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Fermentative utilization of cassava

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fanoansASXTX '
dtilizatxgn
of oassaya
w
Julian Bansson
A Thesis Submitted to th© toiiuat® Faculty
for the Degree ©f
m m m
m
philosophy
Stjor Suh|«©t Biophysical OJwjaiettf’
Miwe&^wA i
’-'--■ .
'’-'-
tea.
la" oli^ge
jor worE''
Iowa State Oolleg©
1940
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U M I N u m b e r: D P 1 2 3 6 6
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B£28f
fa Dr* 1. 1* Pulaier aoft Dr* 1* A* ftiAarkofler, th©
author of this thesis takes pleasure in acknowledging his
Indebtedness for the suggestion of the problem for the
investigation and for the very helpful advise, criticism*
and enseuragsment which they
th© course of the study*
m
freely tendered throughout
flit author also wishes to express
his thanks to the College of Agriculture, University of the
Philippines, for supplying the samples of cassava*
~~7~ 6
f-7 y
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fm m
MffSNQ
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4
HI STORXCAL
* • * «.# - » * * * *
HAffSSXAXS
*
METHODS
,* *.
• • •* *
• *
•. » .
•*
.* * * * *
IS
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m m m m fA L m am im
Alcoholic Fermentation
Studies on some factors affecting acid hydrolysis
and the fermentation of oei& hydrolysatea * * * , *
Studies on methods of thinning cassava pastes . « •
-Inccharifioation by raold-hran
f ..
Saocharifioation by barley malt
Butyl-Aoetonio Fernentation * . . . . . . . . * * «- .
Replacement of corn by cassava
Iffect of adding various materials to. cassava
mash on yield of total solvents . . * ♦ « * . . . «
3 W K A S T ASD CGNCI.TJSIOHS
izTMMTmM Girw
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7
SI
31
§0
SO
81
87
if
90
99
*xm
-4—
I.
ISTRODtJCTIOS
The ©sss&vm is a plant possessing quit® unusual charact­
eristics*
It teas no fcnowa pests m m saeal®#*
It grows in
most soils, resists extrecae droughts, sad' propaga.tss easily.*
It is the cheapest soar®# of starch known*
technologists first m m
When American
©v®r to th# Philippines the- accounts
of tteelr work w#rc filled with eattemslasa over this- pleat*
quote Bacon {1908):
To
*it will furnish the «h#sp#£t source of
starch ia the world and also #0Kp#t* on even grounds withmolasses for the manufacture of alcohol,*
But up to the present
the industries have act mads any us# of cassava except for the
manufacture of starch which is found in-the market throughout
the world bosons# it can. undersell any other starch*
A very difficult local situation has arisen la the Phil­
ippines,
The Islands are foun-* to tee almost devoid of petro­
leum resources*
However, the demand for liquid fuel has
steadily increased with the years.
Is IfSi soot® five million
dollars"1 worth of liquid fuels was Imported*
This was not the
entire fuel bill b#««us# ethyl alcohol, either straight or
blended with 50 per cent gasoline, was and is being used quit®
extensively to power tractors, locomotive#, trucks, and busses.
The alcohol is manufactured from ©arc molasses*
But the entire
solas##* output of the ©ouhtry if fermented would hardly ae-et
the domestic fuel roqulrcoent*
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•5-*
The caasava as a potential source of power alcohol has
been wmh discussed and fermentations oa laboratory scale
bay# testa tried.*
But up to the prmm&t there is no fcaowft
large eoela production of alcohol frost this sour©#*
there
are several reasons for this failure of the commercial d@v#l*
opmeat of the abov® outlet*
'demand,
Cassava starch is still la great
the manufacture of starch is a comparatively sisple
matter compared to the processes required in the conversion
of starch to alcohol*
Starch requires a special treatment,
that of saceharifloatioa, before It can be acted upon b f yeast*
The current methods of 's&cch&riftcation a hi eh employ acids or
barley salt are too costly ant art inapplicable to local
Philippine conditions.
Barley Is not raised la.the country
mad will have to be imported*
special equipment,
The acid process requires
ivem in the Waited States wTh© use of
acids to convert starches for a fermentation process has never
1
bees practicablew*
The fermentation industry is one of the largest consumers
©f carbohydrate material aside from the food industries,
The
diversion of cassava fm m th# starch market would seen to 11©
in its fermentative utilisation,
At present tbs two fermenta­
tions that are teeing carried out on a large industrial scale
1, IT, S. 76th, Congress, 1st. Session,
laboratories; report of a survey,
document 65, p. 31, 1939,
2©glonal research
U* S. Congress. Senate
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—6~
the aXoe&ollo a&& tfe» batjrl-aoetoai# foxaoatatiomu
The
produota of Ifeis* foxmentatioaa, othjrl aleofeol, ooatoao, and
butyl al«oboXf find exteaaiire as«g la the arfca and isdustrioa.
flws Philippines Is aa extremely agrloultaral oouafcaey*
fto#
©ommeroial produotioa of obeuioal* toy f#j»®at*atioa Is a atop
towards iadastrializatioa.
la view of fell# oiroumataaoos presented, t&e foxmoatativo
utilization of eassava was taada the ©bj®©t of this i&veatl**
gatioa*
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XI.
BX3TO&X0J&
fh© oaoeavo plant is feotanieally known no Mauihot
utilisslma PoJiI*
It belongs to t xe fsully Suphorblaoeae*
It is also called tapioca or saaioe although the word tapioca
is often used to desiganto certain foxo* of cassava products*
The plant'itself is a perennial shrub which attains a height
of six to twelve foot at the eg# of one- yeor*
At the base
of its atom it produces a cluster of long flashy roots*
Ac­
cording to luttiiala (1931} the amber of roots varies from
3 to 5 and the average length of the roots is from 27,7 centi­
meters to 43*3 centimeters, and. the disaster, from 4.6 centi­
meters to 7*4 centimeters.
A single root nay weigh 4 kilograms
ant all the roots in m plant nay weigh 12 kilograms.*
{1908} gave m
Copeland
excellent description of the cassava together
with its history, varieties, toxicity, culture, ant uses*
la
the Philippines the yield of cassava was reported by Mendiola
{1931} to fee 16,100 kilo-grassi to 38,800 kilogram* per hectare
(7.18 teas to 17.3 tons per aerah la fmva yields as high as
50',000 kilograms to 55,000 kilogram per hectare {22*3 tens
to '84*7 tons per acre} have been obtained*
Several chcaical analyses of the cassava root and flour
have been published.
Xaaalaaa cassava*
ArcMsoM (1903} gave an analysis of
Analyses by Eeibling of Philippine cassava
have been reported by Bacon {1908},
Oolong {1931} compiled
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til® results of analyses of 53 varieties by the Fhlllppln*
Bureau of Science.
Adriano (1933) mad® a comparative analysis
of flours obtained from ©assay®., wheat, ri.ee, and ©ora*
Adriano,
Bases, ant Tfcalvez (ItSB) found the fresh cassava root to he
composed of §3,80 per ©out moisture, 87.65 par sent starch,
•0*96 par 'cent protein, 1.44 par cent ash, ant 0*01 per cent
hydrocyanic acid*
lytrocyanic sold is a characteristic constituent of the
cassava.
Oolleaa I1914) found the peel of freshly tag root
to contain 0*056 per cent hydrocyanic acid; the palp, 0*053
per cent*
After 3 days the peel analysed 0.245 per cent
hydrocyanic acid and the palp 0.1114 per cent hydrocyanic
acid*
The Increase la the percentage of hydrocyanic acid was
ascribed to the loss of water fey the roots*
According to
Forsook (1937) the toxic principle in cassava is a eyenogenetie
glucoside.
This author found that the processes used for the
preparation of cassava foods do not' eliminate the hydrocyanic
acid.
Collens (1914) however found that the hydrocyanic acid
was entirely removed fey helling, roasting, or drying*
The fatty acids associated with cassava starch were found
fey Lehxosa (1938) to fe® palmitic, oleic, linoleic, and
llnolenie* this last fatty acid was stated to fee peculiar to
cassava starch.
According to this investigator the fatty
acids were combined with the sapless component of the starch*
Data fey Walton (19£9) gave the amylose component of cassava
as 16 per cent to- 17*5 per ©eat, the total fatty acids as
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0.10- per cent, an* phosphorous pent oxide as 0,157 par sent.
According t© this author* "The unusually low content of phos­
phorous and of fatty acids in tapioca starch may possibly be
©©unacted with its reported superiority in certain industrial
processes*
This starch has boos ©onsldered by soma operators
as being the most suitable sours® material for the manufacture
of alJcali-sharcb adhesives* aitrostarohes and eertain typos
of dextrin*w
Studios la other properties of cassava starch war# re-,
ported by Arehbold (1903), by Harvey (19Si), by Ophof |1§56}#
and by Stoma- (1936).
The coaiaerclal utilisation of cassava was given attention
a great many years ago by Swell and Wiley (1@93}»
Moor© (1907)
and bang®' (1909) also pointed out Its industrial possibilities
for the manufacture of glucose and industrial alcohol*
Moor©
(1911) obtained a patent covering th® manufacture of alcohol
from a starch nillk prepared from cassava.,
french breweries
were-employing cassava som @ 35 years ago according to Petit
(19171* This practice was discontinued because of a heavy
import duty placed upon cassava whlcl »as believed to be
eyasogenctle*
Monier-Willlaas (1922), in his monograph on power
alcohol* considered cassava a© one of th® more important raw
material© from the point of view- of yield and availability*
Calculated on th© per acre basis th© data of Monler-Wllllama
plaecd cassava as the highest yieldcr In gallons alcohol* when
compared with ©©reals, tubers, roots, and trees.
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**10**
Collman (1914), at the Ooversuieat laboratory la frinMdft,
eondueted i f w experiments sa the alooJiaXle fermoatatlon of
cassava flour; aslt and taka diastase were eofloyed
eaeeharlfleaticHu
im
ffct experiments war# very brief*.
Th® only detailed mat lengthy investigation m
thm
alcoholic fermentation of cassava a® reported la the literature
la that of Stas anfi Martio (1921)*
The work of these investi*
gators was concentrated on aeii hydrolysis and aiibsetueai
yeast fomentation*
They investigated. th# effect of acid con-
sentratl*nt time ©f boating, sash oeneeatratlen, time of for-
mentation, and th# as© ef stimulants*
Yields over §0 per cast
were claimed#
The alcoholic fomentation of fresh cassava roots was
undertaken by De teen and Valentin (1938}*
Acid hydrolysis
was employed tut the yields were low, the highest being :
©aif
43.84 per cent of theoretical*
Tubsngul and eosforkers (1939} investigated the butylacetonic fermentation of cassava flour*
can replace 90
pm mat of torn.
The
fkey fount that « t m v »
i vtio
of the solvents
butanolt acetone, ant ethanol was fount to he normal, that is,
in the ratio 60:30:10.
They experienced difficulty In fer­
menting. mashes of concentration* above 5*23 per cent dextrose
equivalent*
Sumarirniag, the literature on the fermentative utllissaties of casaava is quite brief.
A detailed study had been
mad# of aleeboiio fermentation using acid saeeharifieatloa*
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-IX*
Cassava was used oaee with satisfactory results in the
brewing industry.
There has been a© work; m
the use of
molds for saocharificatlon.
For th# proooeixtion of th® investigation as reported
in this thesis it m s accessary to review not only the liter­
ature on cassava hut also the literature ©a th# tools of the
investigations
the amylases, mold-bran, starch, and related
subjects..
Th© literature on starch is very voluminous*
Walton
C19£?} mad© m ©©aprehtitisiv© survey of th© field and collected
ji
in hook for* th# liberators i m m 1811 to ISIS.
Fringsheim {1952;} credited Irvine.:with th© discovery of
malt amylase in 1785; a better characterisation of this amylas©
was aade by Kirohhoff in ISIS,. It was found then that the
product obtained frost the act ion of malt waylas© on starch was
a sugar which was. not identical with glucose, and Dubrunfaut
{184?)
proved the sugar-to be maltose*
Th© action of maylas© on starch was recognised to consist
of two stages,
first there was liquefaction observed followed
by th® formation of reducing substances*- Marcher {1678) ex­
plained this action of asylas© by postulating th© presence ©f
two components, a liquefying fraction and m .maltose forming
fraction*
-Ofclsaon (1988) and Huim (1925) conl ributed much
of the work that led to the present acceptance of Herk®r*s
theory.■ According to Hopkins and Krause (1937) the liquefying
oompoBent of salt amylas# splits the starch, partial© 1st©
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-18-
smaller particles with very little formation of sugars*
olenvag® products m m dejctrins*
fh*
The maltose-forming fraction
attacks th© colloidal starch partial® and splits off aoltooo
without causing a market €®©r®aa* la viscosity,
fh® lique­
fying ©ompoaeat -whieh is also known as oC-amylas© is more
resistant to beat than the aaltosa forming component f fiamylas#) *
fh« saooharlfloatloa of starch by
has. been knows
not to yield quantitative osouiits of maltose;
it was found
to stop at a limiting value of' 75 t© 80 per cent of the' sugar*
Pringshtia (1938) considers th® process as belonging to an
equilibrium reaction, th« sugar- already format interfering
with further aaeaharifloation*
Th# us© of malt for sasoharifioatioa. is little known la
th# Orient*
Instead, various fungus preparations are used,
in the production of alcoholic beverages*
The most widely
known of these preparations ar<§ taka koji and Chinese yeast*
This latter is a 'dried rice oakw rich la a nieroflora which
Is strongly aaylolytis.
Sines it is found in many parts of
the Orient, its method of preparation varies with the local­
ity,
In the northern Philippines the procedure is as followss
A dough is aade of steamed rise■and mixed thoroughly with
pieces of an old cake*
The mixture ia laid over young: banana
leaves and set aside overnight.*
The dough partially liquefies
and become® sweetish in taste; later it turns strongly alco­
holic*
Th® liquid portion, which is separated, constitutes
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-ISan aleohoH® beverage.
The solid residue Is pressed Into
rotund flat eaHes and when- dried It becomes tbs so-ealled
Chinese feast e* e
'Lafar Iif 11) gave descriptions of the
methods used by the people in
$wm*
Calmette in X89«s made & study of th# aioroflor®. of 6hine.se
yeast hake*
1#. found a Ktinog whleh be assad Aarloaaroag rongii
always present in his samples aai to this fungus he ascribed
the amyl®lytic activity of tbs eak@».
A process
of alcohol!®
fermentation employing this organ!am was dereloped by Calmette
and Boldin* ’"It is known as the Jay!o pro®ess*
Owes (1933)
gave an- aeeouat of the details of the process* which oon&lsted
of three steps s
CD gelatlnizatlon of the stareh under pres­
sure; (2) saeeharlfleation of the stareh by dlastatle action
of tbs sold; and 15) fezaentation of the sugar by feast.
Th®
advantages derived from the process are eeoncsay due to saving
in malt, decrease in loss®® due to infection Introduced with
the malt, and a higher purity of alcohol found is the .mash.
Also the adaptability of the pro©ass to teoperatures as"high
as 38° 0. stakes it of value in tropie&l eountries*
The trend of development in the Amyl© process is" in the ■
isolation and employment of'more vigorous species of organisms.
In th® Boulard pro ©ess use is mate of Mueor "Boulard lumbar 5*
loidin (1933) reported the aaployme&t of Kuoor iellBiftg as giv­
ing th# best results in industrial application*
Yeasts* like
SaeeharoBuroes aaaaenala* isolated fro® sugar oa&e in Coehin
China* and yeaata knows as Boulard Busbars 81*5© have been weed*
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-14Some of those yeasts are aotive at high temperatures and live
in symbiosis with the mold,
According to Heubatwr (1933) the Aaylo process suffers
several disadvantages*
Th© power consumption greatly exceeds
that of the salt process.
An. expert personnel and a complete
bacteriological laboratory are required*
In the perfection of th® Aaiylo process Boidln (1933)
■■found that it is not sufficient to bring about favorable con­
ditions for the development of the molds*, fh# accomplishment
of diastatic action 'Was also important*
fh© physico-chemical
state of th© wort, the -satisfactory liquefaction, and th©
reaction, were deemed essential*
The fomentation by th©
Amylo process of corn, sorghum grain,, and other starchy mater­
ials were reported by Gall© (1923).
fh© sacchar ifi cation. of starchy mashes for alcoholic fer­
mentation toy Aspergillus orvzae has lately been revived*
This
mold has been used in Japan for a number of years'in th© prep­
aration of rlc© beer, soy sauce, and other fermented products*
It remained for Takamine to bring this sold -to the attention
of Western countries*
In a review of M s own work, Takamine
(1914) traced th© history of his efforts from 1891 on, to pro­
duce an asaylolytic material which would take the place of malt.
Wheat bran was found satisfactory as the substrate for growing
Aspergillus oryzae « As soon as vigorous growth ceased, the
bran was dried and th© product thus formed was called taka kojl.
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Wings taka toji was. asploysMt instead of salt, good yields of
aleobQl wars obtained*
The m m product m e t much 1 m s than
malt and required ©hly one-third to oos-fsurtb th® tin© needed
for preparing aslt«
Tskm&lm {ISM, 1096, 1 U J
1911, 1913, 1913, 1933} took
amaarous patents la the United States far his diastati® prodact a ad Its as# la the fermentation industry*
Osbiiaa and
•Sburah {1933} studied in detail tbs enzyme in taka kojl*
Strains of tbs Aspersdllua flavus-oryga# group, ehieb wore
isolated t m m k©.ji used ia m y asm® .isaamfaotur#,1war© found
to vary in their ability to produce tmyl&ma.
Uvea la soy-
hem. flour which east alas no stareb, strong aaylolytie activ­
ity mma prodused toy aom strains*
lb,® intracellular easyi&es
passed out into tbs culture medium soon after sporulaticm*
Of tbs media studied, which iaelouvd corn a»&.i, coconut meal,
peanut meal, ground dried codfish, casein, and crushed soy­
beans, tbs toast found was wheat 'm m *
Qshioa {1938} extended this study*
®f the
Qiizym
Me found
tbs activity
to to# greatest at pH 4*8 to 5*3 and to to# aest
statoi® to beat at pH. ®,4*
At 35° 0* «nd mm boar heating it
to®#®© completely inactivated*
The preprnmtion and properties of Asoorid.ll.ua mm.m.
enzyme wars further studied by Harada {1931}*
A culture of
tbs sioM was ground in cooked wheat bran containing 50 par
scat water*
It took two days to obtain maximum growth*
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fh®
I
«nsjn&# m m prepared bj alootolle praalpltatioft*
M
inpovtftst
footers for ©msara© production, tt» following w m m listed:
quality of team, water eomtamt &t bran, hydrogen Ion ooisosa*
trmbion, fia# of incubation, teapermtmrs, btiiithXiy of chamber,
and at«ri31nation*
Details, temarar, mara.aot glvea*
la
interesting point la HfcraAm's result# warn the fiotlmg that the
optimum pH for enzymatic activity Increased with laoreasihg
temperature*
was 6+4,
At 30° C, tha optima* pH was §,3f at 65° C., It
1mmwmWf at tamparatturaa below §i® 6* th# o p t i m
pH ramtlaaA praetiaalljr oomstamt at a vmlaa of 5+JB*
Studies m
Aspergillus orama© until very raaastlp wara
oomoastrataA ©a tbs eaaynaa*
Th* original prtpoaitiom ©f
Pakamlaa t© ms# tfea soli-bram itself in the aiaolkoXta faanaaa«*
tatlam process was largely forgotten,
ladarfcofler, fulmer*
uni Sefcoama fliSt}, revived tha idea ani uaftartook a detailed
study of the problem, light species of bacteria and 31 species
a* strains,of molds aara tastaft for mmylasa production*
Molds
of the &hlmamma apaaiaa as well ©a ateaiss of th* . M m m l M m :
flavus-oryza® group wore found to poaaaas high aaiylolytic
activity,
Th# AaaaralHma aaltaraa wava m m m rapid and uni­
form in their growth and attention, was- aoaaantxataA on them*
HoM-braa was praparad in aerated five gallon rotating, pyrex
bottles,
An imfortast oostritmtiom w m th# aapXafaaat of acid,
up to 0-*3 normal hydrochloric acid, Imataad of water*
pravamtad growth of aomtamisuitiag ossamiama.
This .
A. detailed'
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proeednr# ©f the p m m m
wm
develop#!,• Th# prepared mold-
fere® wm «s#f la plrne# of salt- for the saeehartfieatloa o€
o&m mmk*
fft# wmtmk mmmmmf wm found to be 8 to 10 per
e«nt of th.® weight of #©rn*
$&eoharlfloatlo& was. satisfaotorf'
within th® range 48° to it® 0,
ninety per east ©©aversion to
mtmlml was obtained la fermentations of Si gallon® of maeh*
fh® investigation pointed oat advantage# in th® as# ©f a ® M *
bran instead
of
malts
eheapn### of th# Material* high aleehol
yields., and shorter tint® of preparation*
Sehoen# (1939) mat Sshoene, falser, and Uhderkofler (1940)
eosapaxed malt, «©ld~bran, and soybean steal a® saccharifying
agent®*,
if' th#®# th*®#,
soybean least*
was the most effective and
Th© types of aash#«, sofaal mat think, war®
also studied is eonjanetltm with oonhlaatiow of th® thro#
anylolytl# oat#rial#*
1© advantage
tlons ©rtr aoM-fersa ©Ion®.*
wm
found'using ©©sifelii:®-
With a©ii*seo@terifl#i. Hashes,
th® addition of sold-brp© gave eoasiderafeXe iaors&s# la. aloohol
yield,
fitis result was asorlbed to easfsati© actios, the con­
version of + is aon-fermentable poiyeaeobanides twaialsg in
th® aold-saeehartfied nasties to femeatahle sugar,
'fh® greater
effeetive&esa ©f meld-bran ©vsr malt was assoelated with the
greater variety of eaasymee present in th# former*
fh# addition
of soybean meal to: mm14-feran was found to inere®## alcohol
yield® markedly la aoi4-sme@hmri.fled mash#®*
It was suggested,
however, that th# employment of mo14-bran alon# without
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18p a o lls im a n f » *ld h jrd ro ly o lo n ig h t too I t s * A if f le o lt a a i
mm
ooonomieatl*
Bootcaloo (1840) lftv»*tlg«to& rations soli m y X m m
la •
th* ooooiuurifloot Ion of *ov&'aaoh for ottuuiol foxaogkatlon*
four strains of Asporgillao
two
strains of Utlsooms
spoeios, a Kneer, obA two o&ldo&tlflod solit# w#s» grows on
molstonsd wheat fcraa In m iototing arua apparatus.
It
wm
foonA that two otrala* of Bfaitosw* spool## wort- os high saylas#
pro&ueoz* as the rofortnoo a o M iigg#miilt» o m m m *
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~li~
m «
mmwrnMM
Th* m o m iwportaat materials u**& la I M s iavastlgatl**
w m m tli# following $
Cassava flour
Chip* of
and ariei oaasnv* roots war® obtained
froa th* College of Agrl*oltur*t t&ivojpaity1of th*- Philippines,
Philippi***#
Th® chips w m m ground in a burr M i l to a flour#
On# bateh analy«*4 88*1 par o*at glue©#® or 78*2 p«r cent
.
starch; another bi tch aa*lyx*d 91*8 par east glucose or 82 §
per Croat stareh*
sagagi
ia this lanaatlgatloa the tana ground aasaava is used
to designate th® material obtained by grinding th# sliced and
dried uap®#X@i root*
Bee**** th® psel or skin of th# root was
not raocnred, this itatsrial would «ost v**y omoh less than
cassava. flour* .It should be of greater industrial insert****
than' the flour*
A 108 pound lot was obtained
tmm
th® College
of Agriculture, Chiveraity of th® Philippines, Philippines.
Co® bateh of ground ******* analyse** 79*1 per east glucose,
another hatch analyzed 85*2 per o«nt
analysed 82*8 per sent gltteoa**
xucose, and a third bateh
Then® percents### ©orrespoai,
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respectively, to 71*Ej 76.6, aiat 74.5 per cent starch*
Mold-braa
fh* aoM*-fe»B used in this investlgattea was prepared
according to th® oethod of th&sxfcefler, Ptslner, aat Seheefi*
{1959}.
It w*u3 sir iri®i and passed through a Yilejr Mill*
Barley salt
Th® barley salt, obtained from th® PlelsefcaeaB Malting
Company * was ground to a ©oars® powder ia s burr sill*
Bacterial enzyme ”Rai>iaaas*
,
fh# sample of th* feaeterlal enzyme ”Bapidase*
mm
ob­
tained from Dr* Bto&egfcefler of th* Chemistry Bepartaeat of this
College*
**a®pi<ias«w'Is prepared by the Vellerstelh. Company,
Sew fork City,
EslisMssS.
Th® salt eztreet seed ia. this investigation to- prepare
beer wort for yeast cultures was Blue Eibbon Malt attract
eeaoerelally available from th© Preaier-Bafest Corporation,
Peoria Heights, Illinois.
Slsoellaaeous materials
Shrimp powderi
BriaA shrimp was obtain®#, fro® the open
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market in Chleegp*
1% was ground by passing oaea through a
t e f at 11*
Cfempreesed yeast was of the Flalschrnana brand*
Soybean* earn, and m m
grades*
gluten were of the eeistiiereial
The soybean was ground in a toorr mill*
Waters.
Distilled -water was shed la ail «xp*riiseit£s+
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i?»'Miffioas
A*
Klorobiologiaal Froo*&ttr*a
Yeast culture '
One laMfti grams of malt extraet was dissolved la ils»
tilled water to make 1 liter of eolation*
median was placed la Irleameyer flask*:
Such beer wort
30 ml. In SO ml*
flasks for earrflag the cultures, and. 300 ml, in 500 ml.
flasks for inoculating *xp*rlaental sashes*
plugged with cotton
and
The flasks were
sterilised for 30 slants® under a
steam pressure of 18 pounds*
A stock culture of Saaokaraaure** cerevisiae. designated
in this laboratory
m
yeast Hunker 45, was transferred to
a 50 ml. tear wort flask an# incubated at W m C, for 24 tours*
By means of a sterile pipette 5 to 5 ml. of this yeast culture
was transferred, to another flask ant incubated at the same
temperature an# time*, fransfers were mate in a similar
manner, daily, throughout th* course of this investigation,
to maintain a vigorous culture of yeast*
Th* laoonlum for experimental mashes was. prepared b y '
Inoculating SO© ml* of beer wort with 15 to- 20 ml* of a vigor­
ous yeast culture*
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—S3*"
Culturing of the butyl organism
About 0*2 to 0*4 gram of a spore culture in sand of
Clostridium aeetobutylloum designated in the collection of
this laboratory as PCS, was transferred by means of a sterile
pipette of wide glass tubing into the bottom of a tub© of
sterile corn ■mash (6 per eent }. 'The culture was given a heat
shock by placing the tube in a beaker of vigorously boiling
water, allowing it to remain in the bath, for exactly two
minutes, and cooling promptly in cold running water*
tube was then incubated at 37*5° C*
The
After about 36 hours,
when vigorous fermentation had begun, about two ml* of this
culture was transferred by means of a sterile pipette into
the bottom of a new tub® of sterile corn mash and incubated
as before,
the third transfer was mad© into a flask of sterile
corn mash containing sufficient volume to serve as inoculum
for the experimental flasks.
In some experiments, the third
transfer was made into a flask containing a mash of the same
composition' as the experimental mashes; for example, a six
per cent cassava mash containing a quantity of shrimp powder
equal to three per cent of the weight of cassava*
This was
used for inoculating mashes containing shrimp powder.
In all
experiments the butyl cultures were started from one source
which was a sample of a spore culture designated as POS*
inoculations were made from the third transfer*
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All
grsp-a3ratie.it of aold*»ra»
I
fits method of preparing aoM-braa was. that developed by
laisrtoflsr,. Fulmer, and Sehesa# fltit).; fhe sethoi essentially
was as fellowsi
a fivegallon pyrex bottle was charged with
800 grams of wheat termn sat tbo roughly mixed by moans of a.
wooden stick with 1000 ml, of 0.05 nonaa! sulfuric acid*
The
bottle was plugged with cotton and sterilized under steam
pressure for an hour*
Upon cooling to room temperature, it
was inoculated with 80 groats of a well sporoXatsd culture of
Aspergillas orygao* designated in this laboratory as Humber Z,
prepared in the following manners
Sixty grams of wheat bran
was thoroughly mixed with §0 ml. of water contained in a flash
of liter capacity.
The mixture was heated for an hour at 20
pounds .steam pressure*
% © » cooling, it was heavily inoculated
with the stock culture of Aspergillus g | | ^ »
After four to
five days incubation at 30° ..0* a wall sporulated culture was
obtained, which was used for the inoculation of the five gallon
drum, containing a sterilized mixture of
100©
grams of wheat
bran and 1000 ml* of water*
Mixing of-the contents was accomplished by rotating the
drum for a few minutes*
It was then allowed, to remain at rest
for IB hours with continuous aeration.
After'this period, growth
was well started and the dram was then slowly rotated, about one
revolution per minute.
The white mycelial development stopped
and yellow spores began to appear, after forty to fifty hours*
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-£5ffee noM-braa thus foraiet was spread out m
to 4ry la the air.
sheets of paper
ft® air tried preparation was ground In a
burr mill and in this fora wan used for the experimental
mashes.
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—2<J-*
B.
JUutlytloal Frooo&uroa
Determination of starch and of
mmmm
Starch was determined toy noli hydrolysis is ac cor&anoe
with the Official and tentative Sethods of .Analysis of the
AnsoeiatioB of Official Agricultural Gheminto#
The reducing
substances famed la tie hydrolysis were estimated according
to the method of Shaffer and Hartmann (1921).
The reagents
were standardized toy means of a sample of pare glucose*
All experiments in this thesis were earried out in
duplicate or ia trlplloate#
Determination of ethanol
The entire volume of fermented mash or « measured aliquot
of it was distilled and the first 10© ml* was collected.
The
volume distilled was kept below 300 ml# for preliminary runs
had shown that with such volumes the ethanol was recovered
quantitatively la the first 100 ml*
About 0,5 gram- calcium
carbonate was added to the mash to neutralize acids present#
The specific gravity C2#/E5®) of the distillate- was determined
toy means of a Oheinomatio Weetphal balance 'and the ethanol
concentratlan in graume per 100 ml# of solution was read from
a table*
Temperature was controlled In the following manners
the distillate was coaled In'running water to about 24° C.;
on pouring into the specific gravity cylinder the temperature
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-87-■
was raised to exactly 25® C. by warmlag tha cylinder with the
hands and stirring by gentle notation.
the specific gravity
float acted as a stirrer.
Bcteriilnatien SL mm-rn&ti&m t m immwXmi* aoM»bra&.'and. malt
ft® inoculum for the alcoholic fermentable*: was.. made .of"
a solution of malt extract.
’Ydan inoculated with, yeast as
amount of alcohol was' formed and I M i was- the correction for
inoculum. _However* it was found that la th® presence of aoltbras or malt*, a larger quantity of ethanol was obtained# ■'A
differential ttethod was usad to evaluate the corrections for
th® beer wort and the mold~hraxu -for beer wort the calculation
was as follows:
4 g. iaoM»bran + 200 si*, beer wort
#*14 g*- ethanol
4 g. sold-bran + 100 atl«..beer wort ~ 5.15 g. ethi
100 ml. beer wort = 3.01 g. ethanol
fh e .average of two determinations was 100 sal, beer wort
« 3.S3
m*
ethanol,
the direct fermentation of beer wort gave
100 ml. s 2.01 g. ethanol,
for ttold-brcn, the calculation was as follows:
8 g. »©ld«bna» +
200
ml.
bear wort «6.29g. ethanol
4 eu seM*»bran -4-
200
ml.
beer wort s6*14fc. ethanol
4 g. mold-bran
s
0.15 g#/-»t£umal
fh® average of two debexminatieBs was 1.0 g, mold-bran
» 0.0325 g. ethanol* ■froa th® direct fermentation of
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-88meld-bran, 1 g. sold-bran * 0*0120 g* ethanol,
Similarly,
for barley malt 1 g. malt « 0*354 g, ethanol*
Calculation ef ethanol yield
In an experiment the original mash eonteiaed the follow­
ings
40 grass cassava with a, glucose equivalent of §5*2 par
cant;
20
ml* bear wort as inoculum| and 4*2 gratae mold-bran*
The entire fermented mash |300 ml*} was distilled and the
first 100- *1* dlstillete collected,
The specific gravity
f25°/l50} of the dlstillete was 0*9753 corresponding to ,,15.22
grains ethanol par 100 ml* distillate*
m m ms
follows-1
for the
B0
ml. inoculum
for the Qiold-brajB.1
« 0*500 gratis ethanol
4*2 s. 0*0325 s 0*135 grams ethanol
Total
Therefor®, the quantity of ethanol
- 0.74 gr'-ns ethanol
frm
grams minus 0*74 grams -or 14.48 grams,
c#
The ethanol corrections
12g6 — >
the cassava is 15.22
from the equation
200g -f 20glgOH, ISO grams of glucose would
yield 92 grams of ethanol.
Forty .grass of cassava should then
yield
(40
x
0*858} x 92/180 = If.40 grams ethanol.
The ethanol yield is therefor# 14*43/17*40 » 85,1 per cent*
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of total solvents
in
the butyl-aoatoats
tation
The i m a m nted butyl aasfe {the entire valuta© or an'aliquot
if it is larger than 300 ml,! was distilled,
The first 100 ml*
of distillate w m collected a M its specific gravity {£5°/250}
detexfeiiiisd* 'The total solvents were ealouieted fro® the
Cltrl«tszuw&*<fulnMkr equation {1935}«
Total solvents,
gvaws/luO ml* « 698
(1,0000
** *p, gr*
85°/8S®}*
Correction for Inoculum In the butjl-aeetoaio fermentation
was detexalned by actual fententatioh*
.for ejunple* Sf© atl* of
a oorn inseultKi yielded S*0# g r a s of total solvents| a tea ml*
inoculum therefore was equivalent to 0*137 gras solvents*
Correction for additions of shrimp powder, soybean, pep*
ton#, eaa$reesed yeast, urea,.*»# sera gluten
w@m
evaluated by
determining the Increase in total solvents produced by adding
a
kaowa mmmmt mi the
material to the corn mash-*
for example*.
21 grams corn plus 1 gram shrimp powder yielded 4+06 grams
solvents | 81 gran® eera alosa yielded 3#.#© grams solvents*
Therefore one gram of shrimp powder eras equivalent to 0*86 grams
total solvents*
Tim
corrections par great of material found
were 6*26 gras for shrimp* 0,48 gram far peptone* 0*082 gras
for compressed yeast, and m m
for urea, eors gluten* end soy­
bean*
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-30
Calculation of yield in the Imtrl-aestoaie fermentation
Tlald in tli® butjrl-acotonio formont&tion was oaloulatad
ms grama total solva&ts pay 100 grams glaeosa aqalTalsnt*
For,
sxampla, from 21 grams of easaava having a gLuoos*' equlvaie&t
of 17,4 grams., 4. ft grams total sol vasts ware attained after
daduotlons tad feeea mad# for tnoonlwa.
Tbe yield was therefor®,
(4.19/17*4) r 100 * 27*4 grams solvents par hundred grams
glucose in the sample*
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v* sxptimsifja m sm s s
A.
.Alcoholic Fomentation
I# 31tidi.os cm. 1101110 fhetors sxfog 1 xng iili fetgsatola .m.. M M
fermentation of noli hydrolysutes.
The oonvoreIon of starch to glucose by acid hydrolysis
lias boon used for years on as ti dust rial scale.
Thor® have
been aunoroaa reports is ih# literature on the ootpleyiBeBfc of
acid oaceharifleatiozi in alcoholic and other fernantatione*..
Those studies have boost only on laboratory scale and
m
far
as knows the alcoholic fermentation Inu-iTfcry docs act find
the use of acids feasible.,
Jtsverthaleos, data on the behavior
of ground cassava root towards acid hydrolysis were deemed
desirable.
a. Iffact of acid concentration and the .ratio samples
acid m
the lierefaction of aaeh ami on the per cent conversion
to SSISIJI*
fipww* of ground easeave. root were thoroughly
mixed with sulfuric acid solutions.
ware.:
The concentrations of acid:
Q.l, 0*2, 0*4,-and 0.8 normal; the ratio grams sample:.
ail, acid solution was varied aeeor&lng to the proportions of
1*1, 1*1.5, 1*8, 1:2.5, .and 1*.3;- for example, 10 grasm of
sample wore mixed with 30 si* of sold solution to sake a 1:5
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ratio*
All the mixtures wore heated under a steam pressure
of 20 pounds for two hours.
the samples was noted*
The degree of liquefaction of
If the sample was definitely liquid
and flowed at room temperature it m m designated -t- In Table I;
if it ©afced solid end appeared dry, then it was designated — •
Sufficient water was added to the. flasks to dissolve, or
suspend the hy&rolysate and the solution diluted to erectly
100 ml*
Sugars were determined from this solution by the
Sfaaffer^Hartaana method,
The data are ©©Hasted in Table I*
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W !
I
M t m t of Acid Concentration; « 4 the Ratio Samples Jolt on
Lt§u«f&&tlon of Mash and on the for Gent Conversion to Sogars
Acid
0*1
0,3
0,4
0*8
Samples Acid
{Dilution}
Killl©tplvalent
Acid
Lique­
faction
% Conversion
to Sugars
XtX
1
—
not determined
1*1*8
1*5
—
m
XtS
Z
—
n
li8*5
2*5
—
n
ItS
S
+
«
111
2
—
*
Is1*5
3
—
*»'
1st
4
«
ItS* 5
S
±
+
19*8
1*5
8
+
38*8
111
4.
—
1:1.5
6
±
33*4
111
3
+
75.4
1*3*5
10
+
98.0
its
11
+
94. S
111
8
±
§8.3
1:1*5
11
+•
87.0
111
18
f
90*0
Is1*8
88
f
94, S
1;3
14
t
08.0
not determined
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-34this esperiaent was carried out to da'texKine the limiting
valus of tli#
sugars*
sample?- acid for satisfactory conversion to
The results obtained indicated that sulfuric eeld
concentrations of 0*1 and 0*8- aora-al were not satisfactory*
With 0*4 a o m & X sold, 93 par cent of theoretical glucose was
obtained when the samples. eeld ratio was lit.*5*
W m i tbs moil
eoaaeiitratlott wee raleed to 0*8 normal» 8? par eent of theo­
retical glaeos# »as ©btmiaed at a sampler arid ratio of IsJUi
and ooa¥©rsi©»s of 90 per cent or above were.-obtained at higher
dilutions*
At equal quantities of sulfuric mold, the higher dilution
( I m m samples aoid ratio) fevered higher eftdeharifieetion*
The
conversion from 0*4 normal noli and lit dilution was f8*4 per
sent which is higher than the raise obtained with 8*8 m m & 1
acid and a dilution of 111 although in both eases the total
quantity of unit was 8 mllllequlvolants*
Again with 0*4 normal
acid mid 1;3 dilutio * the conversion was higher than that with
0*8 m m m l m i d and lsi*S dilution* the quantity of acid in
both cases bexn^ constant at IS millie<iuivalents*
It is to .fee noted also that 10 allJLiequlvalcute from a
0*4 normal aeid gave a 93 per cent conversion while IB allllequivalents from 0*8 i « l
acid gave only S? per cent eenver-
eioaj even IS milllequlvaleifts of the latter m M
eoneentratlon
did sot give quit# as high.a result ee the first mold ooneentr*tlen*
that the higher dilution favored hydrolysis was alee
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-3 5 -
borne oat by litter experiments*
b# Iffeot of aeid concentration aad of the ratio gaaplei
aeid on per eent conversion to swears and to ethanol*
Using
the preceding. data ms a guide is selecting suitable samples
acid ratios, and employing t b « a a M procedure, the experimemfc
was repeated.
The weight ©f sample was increased to 30 grass,
the eotiking pressure to 15 pounds, sad the cooking time to.
two and. oae-Judf hours.
The hjrdrolysstes were diluted to
g-00 ml* and Z ml* of the solution wee pipetted off for sugar
deteraiaatioBS.
to the aula volume of hydrolysate fl§8 ml. ) a.
sufficient quantity of concentrated «aBoaiua hydroxide was
added to bring the pB to 5*
yeast culture,
laoeulatloa was made with 10 *1*
After luoufeatlos at 30° 6» for seventy-two
hours the eloohel was detexmlmed*
Is la the preceding experiment the estooat of sugars actually
obtained by the hydrolysis was compared with the theoretical
aaomat derivable from the etareh*
in Table 11 as
The eaaparl*e&» are expressed
eonversien of starch to glucosew*. The actual
weight of ethanol obtained by fomentation was also compared with
that which would have been obtained from the glucose present
had the conversion been quantitative; the values m
calculated
are recorded in Table II under th® heading m-$ eesversioa of
glucose to It 08**
Similarly the per cent eottversioa of the­
oretical glucose to ethanol mmn miss calculated*
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R eproduced with perm ission o f the copyright owner. F urther reproduction prohibited w itho ut perm ission.
fable II shows ilal under the conditions of the experiment satisfactory conversions of starch to sugars war® Oh~
•bal&ed with 0*4 normal sulfurl© acid at sample:sold ratios
of ls8, 1:2*5,. ■and;1:3 and with 0*8 normal acid, at propor­
tions as high as 111. and 111*5.* The advantage of using as
high a ratio of sampl® to acid as possible, is economy of
autoclave space espeolally whea th® autoclave has to be of
especial material resistant to acids*
Is in the previous experiment* it is shown by the data
in fable II that at equal quantities of acid the lower rati©
of sample:acid favorsi higher saocharifieatloa as demonstrated
by sample nun&ers f and 10, and 10 and 11*
However, this was
hot always the case,, for sample numbers 1 and 5, and sample
ambers 2, 4, and S exhibit the- reverse tendency.
Both con­
centration of acid and'dilution of sample seem t© be important
■in saccharlfi'cation by sulfuric acid*. Sigh acid concentrations
prenote hydrolysis ©f the starch to sugars but have ® tendency
to destroy the sugars formed*
High dilutions of the sample
prevent destruction of the sugars bat slow down hydrolysis*
la the' present experiment an acid concentration of 0*4 normal
and dilution of 1:2*5 or 1:3 showed the highest degree of con*
version of starch to sugars*
The conversion of the sugars "(formed by the acid hydroly­
sis) to- ethanol by fermentation did not run parallel to the
quantity of sugars present*
Thus sample number 6 with only
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61.3 par cent hydrolysis gave 'the .highest ethanol yield of
81*4 p f e«it
this result is in aeeord with ths findings
of Severson <1937) who worked with th« acid hydrolysis of
©ora*
The par seat eoaversloa of theoretical §ia@®s® to ethanol
\
s«®s«i to l&erease m
tho ratio- sample:acid decreased.
This
was invariably ths ease irrespective of ths acid m m m n t m *
tion*
Thus sample numbers 5, §» 7* 8, and i show a progress*
if® increase of per seat conversion of theoretical glucose to
ethanol*
* * S S ssE m i l i i l H iM f it .s iH a ii la s it iE figgaalft a t
aold-braa to sold hydrolrsatc.
thirty grams of cassava floor
war# thoroughly mixed with S50 ml. of -§♦! normal sulfuric acid,
gelatinised by heating over a burner, and autoelaved for two
hours uador'a steam pressure of SO- pounds*
Hpon. oooliagt ths
pH was adjust®* to 5 by sieans of conoantratad .ammonium hydrox­
The sash was saooharified with various amounts of mold-
ide,
brah for on# hour at 6©13 G. and thea was Insulated with
yeast*
Alcohol was determined after iaeubatioa at 39® C*
for fifty-two hours*
The results as shown in. Table 111 indioat® that the add­
ition of isold-brea increased the alcohol yield.
This is in
accord with the finding® of Schoeue {1939} and of Sehoe&e*
Fulmer, and t&derkofler CltdO) who worked with earn*
tftt&er
the eoaditioAS of the preseat experlae&t, the quantity of :
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-ituoM-braa required to i&orease alcohol yield is about 10 per
e#at*
la. 1a ter •xpovinoats
mmtimimtmf
fields were obtained
with atold-bres elen# without recourse to the m m of ueld*
M i d hydroly#is, under tbo eoailtioas adhered to la the pres­
ent «psrlMat, offered a© adraatag* orer straight a o M
ssoshorlfiestloa*
m i
nx
Effect ©a Alcohol Yield of Adding: farying Asimi&ts of
loM-braa to Acid Sgfdroljrs&ts
Kold-bran, Par Cant
by Weight Sample
Par Gent
It OB ■
0
74,4
1.7
7t.a
5
?#,4
m
7t,S
•
£0
ew'W
81.6
*• S f w l «a *Sfc«a®i z i ® M °f addin* asld-brea. barley
Halt. inactivated taold-bran, iaaetlrated barley aalt« wheat
bran, and yeast extract to meld fardreljrsstss.
the jftrttoi and
qiia&titits of Materials ussd la this «jrp«ria@nt wore tbs
same as in the preceding ©a©*
©tiring sacebariftcation, however,
different aatsrtmls war* added to tbs samples,' tbs seM-brayo,
and barley amlt m m ® lassttvstsd by mixim
tbs* with water and
boilin.ft for fifteen aismies*
R eproduced with perm ission o f the copyright owner. F urther reproduction prohibited w itho ut perm ission.
*»4G"*
fit® 'rmnltB m
given in fable IT indicate that the
fuaefcioii of m o M ~ W m is that of m saccharifying agent.
It
seemed to servs as a eoureo of nutrients ©aly to a slight
exteat because the addition of inactivated mold-bras raised
the alcohol yield somsfvliftt;. so did the wheat brim..
Inactiva­
tion of soM-braa ant umlt lowered yield**
TABXX If
ffiffee* on tbhaitol Yield of Adding M©14~bran, Barley
Malt, Inactivated Mold-bran, Inactivated Malt, Wheat
Bran, and Yeast 3xtract to the Acid Hydrolysate
Material Added
Material Added,
.
$ by weight of 9wm\m-
non# ( m a t m l l
Far Cent
It OH
f4,4
aoM-braa
BO
Iliactlrated mult-*feram
20
barley salt
m
68,0
m
■«4,f
Issstlvatsd barley
malt
81,4
.
??,!
?.§■
ff.4
10;h Mold-bran)
2*5
80.0
.yeast extract (&s&
10$ mold-bran)
31.#5
wheat bran
yeast extract {ant
«* M £ m l s& tMsssi iSjM s
tloa of acid kvdrolygates,
i s m x im M & m s k M m m m im r
Weights of cassava flour -{IS, 30.,
45, and 00 .grams)- ecuresponding t© #, IE, 1®, and 14 p*r cent
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-41sashes were mixed la 500 ml* tlasts with 250 ml* of 0*1
normal acid.
The mixtures ware gelatinized by heating and
then, hasted. under a steam pressure of 20 pounds for two and
one-half boors*
Vp&n eoollng the hydrolys&tes were ad­
justed to pS 5 fey means of concentrated a*i*@iiio» hydroxide
and inoculated with yeast*
Incubation was made at 30° 0.
f o r •seventy-two hours*
The results as given In fable ¥ show that the highest
alcohol, yield, was obtained with the lowest mash concentration
studied*
This is in accord with the results of the experi­
ment recorded, ia fable 111 the higher the dilution the
greater is the conversion of theoretical sugars to alcohol*
The Metier dilutions studied in the present experiment may
account for- the higher conversions*
With a 0 per cent mash,
which would give a saxlau® of only 2*0 per cent bear, the
actual conversion was 81*3 per sent.*
Proper dilution of acid
hydrolysate seems to furnish ms high alcohol yield as that
obtained with aoM-brim.
The dilution necessary is so low
however as to be of a distinct disadvantage. In large scale
fermentst ions.
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*48W 1
?
Effect on Ethanol Yield of Tarytag the Hash Concentration
of laid Hydrolysates
Kash
Concentration
t * M im
Ratio
Samples Acid
Per Cent
Et OH
lflt.8
81*3
in
ls8*3
- *8*0
m
l?§*§
98*4
24
It4*8
48*3
i £H g*sssg& mM M i£ sgJMlii. gaslgaa ztasM
in the fermentation of acid faydrolygatea containing mold-bram*
Thirty gra®m of mmmmtm flour w m m heated with 250 ml* of
0*1 normal m i d for on® hour at 20' pounds eteea preeeure*
Upon oosllag the samples were adjusted to pi 5 by means of
aame&lum hydroxide &»4 treated with $ g r a s of mold^bran for
one hour at $0# 0* ,The- mixture was then eooled to 30° 0*
sad inoculated with yeast*
Toasts designated as amltfi 2,
16, 81, 35, and 43 were employed.
Is shown in fable ¥1, all tiss yeasts studied, except
amber 2, gave alcohol conversions very close to or'above 80
per -cent*
turn highest yield was ebtaine& from amber 43 which
is the yeast used throughout this study*
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fjysu u
Effect of Employing fai'ioms feasts on Acii By&roX.jrsat*
Containing Mold-bran
Y««»t Stab*?
Z
Par Cant
St OH
?0.8
u
79*8
Zl
79.0
■35
80«6
43
«U8
'
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-4 4 -
&* kTfsst on ethanol yield at varying the aeiC-aoaeajtttratloa
and the oookir#, praasure*
fifteen grama of eassava flour were
geletlnlsed by heating with *80 ml* of aeid aelutloas
ious oonoantrations*
Th# cooking pressures as@4 were 0 (atmas*
pherle pressure)„ 15, m A 25 pounds.
vsrie&s
mt var­
The eooklag time was also
15, '3©, and 80 minutes {at IS and 25 founds a teem
pressure); sad SO, 120, and 180 minutes {at etmespherl* pressure)*
Cooking wmm considered started ten aixmtee -after the autoclave .
hat reashed the desired pressure*
gfeesm was turned off after
the desired heating tins heft elapsed sat the autoelere was
cooled as rapidly m
pass111# by a ourrest of air*
Twenty-five
minutes were refaired to taring the pressure deem to etmospherle
without eattsiag the samples to boil over*
The pi of eash mix­
ture was then adjusted to- i by mease at ammonium hydroxide*
Saoeharlfleattoa was carried out at.iO® c. for
m
hour with
soM-bras equal to 10 per eent by weight of the sample*
The
mash was seeled to 30° i* sad inoculated with 80 ml* of yeast
eulture*
hours,
Alcohol was determined after Ineahetlon for sixty
fhe results are tubulated Is Tables fll, fill, sal JX»-
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HIS**
fABLS 111
Effect on Ethanol Yield of Acid Concentrat-loa ami Time
of Cooking (at 25 pounds)
Acid Concentration
fia# of Cooking, Minutes
{Normality)
(25 pounds)
15
m
75.5
78.5
78.5
0.02
71.5
75.5
77.5
0.05
79.7
eo.5
84*5
0.10
S4.0
84. 5
86. 5
; 0*0
.
50 .
TABLS till
Effect on Ethanol Yield of Acid Somcsatration and Yin#
of Cooking (at 15 pounds)
Acid Concentration
fist# of Cooking, Minutes
(Normality)
(15 pounds)
15
38.
50
0.0
80.5
86.4
79*6
0.02
81.0
84.5
74.3
0*0-5
85. S
83.8
77.0
0*10
85* 8
■80*0
76*0
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-*46**
TA B U IX
.Effect on Ethanol Yield of Acid OonssatratlOa sat fiat
of Cooking at Atmospheric 'Pressure
Acid Concentration
{Normality!
Time of Cooking, Miautss
(Atmospheric Pi?sssurs)
60
180
0*0
80*0
@4*1
0*1
85*4
85*6
84*1
0*1
¥8*4
79.6
84.8
0*5
81*0
80*1
83* 6'
180
.
83*6
At a cooking pressure of tw#»ty-five pounds {Table ¥11},
alcohol fields Ioprovad with Increasing acid concentrations
ant iaereaaiag ©©©king times.
This is readily •xplftlaafelu
for both of these factors p m a & t m hydrolysis of the starch
to fermentable sugars.
When the cooking pressure was fifteen
pounds the results were not so uniform*
Alcohol yields in­
creased with acid concentration only when the cooking time
was fifteen Mantes! with thirty Alautes cooking the trend
wag the opposite*
With sixty ailautes cooking, the yields
were comparatively low*
When the cooking was done at atmospheric pressure the
results did act indicate any marked trend, although alcohol
yields attained a quits uaifora value when the cooking time
was prolonged to on® hundred and eighty minutes.
Cooking at
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atftosp&erio pressure seemed to give as good results as
pre*sura beating,
la generel it may fe® concluded that the
conversion of eassava flour to alcohol under the conditions
studied, is a process la which the eoo&Lug pressure, eeld
concentration, » M seeking time are not the ©sly controlling
factors.
h* Iffeot on ethanol yield of aeid concentration and
s
s
a
M
s
s
.titnim
* th
ec
o
o
k
ieM
m.M
4
ms
m
ia
a
i. il su&
s
a
i>
The results reoorded In Tables 711, fill, and IX are not
directly eemparable because the srpeximeiita war® made on
different days*
to study the relation between cooking pres­
sure asi acid ionoantratiea, runs war® made employing the
same materials and methods of the preceding experiment*
Mashas of different acid ooneeatratloss war® cooked at the
same greeeure.
For example, mashes having acid concentration®
of 0*0, 0*05, .and 9*1.normal ware steamed at the same uniform
pressure of 15 pounds*
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>48**
fJkSLS X
iffect on ethanol Y i d d of A d d Concentration m A Cooking
Pressure; Tim© of Cooking One Hour
Acid Concentration
(Normality}
$©okin« PrOBMVOPi»• Founds
.
.
0,©©
0*05
'
0.1©
0
15
25
81*4
73,4
*5*4
*5.2
77,7
*5.5
80*0
'
*8.4
84.5
According to the results gives is fall# X* at cooking
pressured of 15 and 25 pounds, the trend was for improved
ethanol yield m
a d d concentration was increased.
then the
cooking pressure was atmospheric, the results were rather
irregular,
This- any he due to a factor which was feotag elia»
lasted hy higher cooking pressures and stronger a d d solutions.
.©s. the whole these experiments show that eti ansi yield#
of m m
BG per ©eat can he ©htnined from. the aleo&olle f«r*»
aontation of eaosava without roeoureo to pressure cooking or
a d d hydrolysis.
1. Coap.aylson of alcohol yield from cassava flow? and
from ground caagave root, a d d hydrolysed at varying pressures.
Fifteen grata saiapXes of oaggttva flour and of ground cassava
root were a d d hydrolyzed as la preceding experlaests with
0.1 normal a d d for ©a* hour*
Cooking pressure® of 0- (otao**
pherle), 15., uni B5 pounds worn studied.
©pen cooling the
a d d hyir©lysates ware adjusted to pH 5 with concentrated
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•49.
ammonium Hydroxide.
K«M-braa equal to 10 par cent by
weight of sample was aided and the sash was inoculated with
yeast,
Alcohol was determined after incubation at SO0 0.
for fifty tours*
The results nor® eammarlaed la fable XI.
f ABX1 U
Comparison of Alcohol Yield from Cassava Flour and from
Ground Cassava Hoot, Acid Hydrolysed at Fary.tug Cooking
Pressures.
Pressure. Pounds
Sample
0
15
18
Cassava Flour
*3.3
80,0
84.5
Ground Cassava Hoot
78.3
85*0
86,0
The above data show that the entire cassava root, peel
and all ground together> is a hatter' r m material for alco­
holic fermentation than is cassava flour.
The alcohol yields
were Invariably higher with the ground root than with the
flour.
This may he partly dm# to the higher percentage of
protein in the root*
Archbold (1903), in his analysis of
Jamaican cassava, found the whole root to contain 2*45 per
cent protein and the flour only 1*§1 per cent.
Senior-
Williams f19221 also states that fermentation of concentrated
worts are assisted by the presence of a certain amount of
solid matter*
The hark particle® might be performing this
role*.
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■•50Besides offering. improved fields of alcohol the ground,
root is auoh cheaper than t&e flour,
A large part of the
m st ©f preparing cassava flour -seises from tbe peeling of the
roots,
Is far as known ground eaaaeva root has not been used
in the industries nor studied in the laboratory.
**
3M M m m
a s M M f.
st I&
la lis . a im m is s ls s *
Meshing may he defined as the process which renders a
starchy material ready for alooholle fermentation,
cess may be considered to involve three steps:
the pro­
gelatlnlantion
of the starch, liquefaction of the starch, and saccharifies*
the final objective Is- a soluble transformation product
tlon,
which can be readily acted upon by yeast*
low sash concen­
trations of cassava do not present much of a probleaj the
higher eoaeeatratloas offer difficulties mainly due to- the
■thick* muollaglnous pastes obtained,
desirable because of resulting
Oomeantrated .sashes are
mommy of space
of cooker*
feraenter, and still,
a.
OelatinlelaK temperature, In a SS ml, Srlenmeyer
-flask, S grams of ground cassava root were mixed with BO mi.
of distilled water and heated slowly in a water bath.
The
temperature of the bath was gradually raised, the- contents
of the flask being, stirred in the- meantime,
fh# temperature
at which the mixture stiffened to a paste- was taken as the
gelatlaissiag temperature,
'fhis was found to be about t0° 0,
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-§ t»
For tfa® purposes- of aaafelitg the «bov# value was eoastt#r®t
stiffioieijtly oocurata b»#at»a all tt»t was desired was the
approximate temperature to -which a cassava-water -sdxtmr®
must be heated to aa&iev* galatiaisatloa of tto# starch.
low value of tht* teiiperabmre is of aigalflaaaea.
Thm
ffe® starch-
liquefyiog ooaipoaaiita of *ooa asjrlaaaa m m still active at
70° C. although for only a abort tin®,
Asyl&s*® are known
which are capable of attacking staroh avaa at S®° 0.
For sag#
of sashing it would, b® daalratla to bar* m gslatiiiiziag teapsr*
atur# low eaaugii not to inactivate aaqrlaaa or to locate as
amyla**, aaffloiaatly resistant to best, to atbmtk starch at
or above Its gelatinising, tsapsratar®.
Too general procedures
for the- llquafootioa of «taro& atay be followed:
First, the
aoylasa any be heated together with tba starch land water) up
to the gelatinizing t«p#ratiire; the aoyXaM would attaalr the
staroh granules during the proses® of swelling and the starchwater mixture would be expected to hardly pass through the
gelatinized state at all,
Second, the amyl&sa say be added
to the .starch at a temperature above it® gelatinizing temper­
ature.
fhese procedures are designed to avoid carrying out .
of the liquafylng operation at toBparaturo® lower than the
gelatinizing temperature feaoatiaa then pastes encountered are
thick and difficult to work with*
It was noted, however, that cassava paste® vara fairly
workable at #©0 to S§0 0* whoa they w @ m u m & at one® and not
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•52allowed to 0001 down to m m a temperature.
b.
jgl of 'cassava mash*
h m & a pS of about ?*
Cassava mashes were found to
As expected it varied-from sample to
sample, but only slightly*
Thus.-30 grama of cassava flour
gelatinized with. 250 ml* of distilled water gave a pH of 6*®*
Ground cassava root under similar conditions gar# a pi of
7*0*
To ietaraiB© the amount of a d d required to obtain’a
desired pi, the following experiment was serried out:
To
96-gram- .portions of ground cassava. root, §50 ml* of sulfuric
aeid solution, containing-a definite, valuta®- ©f 2 UQtmml
sulfuric acid, were added mat the mixture was heated*
The
gelatinised mixture thus formed was autoelaved for forty
minutes at *0 rounds steam pressure.
The pi was determined
upon eooling to- room temperature <26° to £?° 0).
'T4BLi 111
The pi of 'Qelatluised Oassava 0-oatalttiag Varying founts
of Sulfur!© Aeid
ml. 2 sormal -%S§4
car 25® ml*. .
PH
0
7,07
1
§*55
2
#,1®
3
5*28
4
4*as
R eproduced with perm ission o f the copyright owner. F urther reproduction prohibited w itho ut perm ission.
53**
o. Saeeharlfleatlon at 55# to 60° £.
were found to tea difficult.to
mask
Cassava pastas
at 05# to 60° 0#, the
usual ..saeoharlfylng temperatures, too to the stiffness of
pastas,
tkm
fo obtain a final alcohol concentration ©f 5 per
cent, the original cassava Mash must teavs at least IB grass
of material per 10C ml* mash.
la even higher
ttos would be advantageous and desirable,
To
mmh
concentre*
overcome or
avoid the formation of thick mashes, the operation known as
thinning was gives careful study,
A* ^bisuing with acid.
When 40 grams of cassava was
©coked with BOO ml, of 0*1 'normal sulfurlo aeid for one hour
at 20 pounds pressure» the result was a this aash.
Using
0*05 a o a a l aeid instead of 0.1 normal aeid, the resulting
produet was not
mo
fluid, tent was sufficiently thinned t© tee
workable.
Attempts war® also made t© employ sulfuric acid concen­
trations as high as 0*8 normal.
trations, the ratios
up to lil.
of
At these high acid
concen­
sample to acid studied were increased
Using 10.-grams of sample and autoelaviag for eat
hour at 20 pounds stems pressure, the rati©, of sample to aeid
that gave a pro&ust that was more- or less fluid end not naked
hard were the followings
with 0,8 normal acid, 1:1; with
0,4 aersial aeid, lsl.S; with G*£ aoraal acid, ill; and with
0.1 normal acid, 1*3*
the produe't from this aeid treatment, was
diluted with water and formed a rather thin sash,
On© objection
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*54*
to th® employment of as 14s Is the necessity of neutralizing
the 6X6983 aeid,
9* Thinning with dry s M s S S S *
With 40 grams of
easeava was nixed thoroughly 0.0 grams of dry aold-braa
{2 ,p®r cent of weight of staple),
Two hundred ml. of water
at 85° 0* were then added with e©&binti#<t .stirring of the
paste formed« Th® teatperature of th# mixture was a little
above 70° 8., whieh is the gelatinizing teoperatare.
The
mash was at first thick but llqasflet sufficiently upon
standing for fifteen to thirty minutes*
Mashes
m
concentrated
m
.24 grams per 100 ml* of mash
were prepared with this method of thinning and found subse­
quently to give good yields of alcohol.
However, 14 per
cent mash is about the satisfactory upper limit with regards
to ease of working,
for the thicker sashes e larger amount
of thinner could be used with advantage,
la experiments on.
aoM-bran* it was found that the amount of thinner could be
increased to 3 per cent of the weight of the sample without
detriment to. the alcohol yield., although the mount of mold-
bran for sacsharifieatten was reduced he ? per cent,
While
this method was found to be very .satisfactory with reference
te the preparation ©f mashes, it suffers the disadvantage of
losing s good portion of the active amylase of the sold-torsn
through the seeking subsequent to thinning*
la this method
the mesh is first thinned and the seeking 1* performed later*
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*§§«.
** Thinning with moM^bran suspension.
Ttttossi&s* Inc.
m®mm®n€ th© ms of Q.5 -for ©cut by might of' sample of their
beica-koji (a i&sberlai similar to m©id«bra»} for thinning, th©
koji being a*Ae into a asspsftslofi in water.
In line with
this procedure, oxperiwintft were tried with isold-hras Suspen­
sions. ■fo 100 ml* of distilled water warned to 40® 0-*., 1.8
. grass of mold-bran wore added and shaken vigorously.
The bran
particles settled to the bottom of the container; th© super­
natant liquid was light g v m s l t h yellow and somewhat turbid.
Ton ml. of this suspension was used to thin cassava past©#
in the following auuutaces
Th© heated and gelatinised cassava
past© was cooled to 74° C. and at this t«f®r»tiir©* a volume
of th® prepared thimner saatalnlng a weight of stold-bmn
equal to 0.5 per cent of th* weight of th© sample m s added
and th© mixture stirred,
place speedily.
liquefaction was found to take
Will© this method was found to be effective,
It worked best at mash coacentrations in th# neighborhood of
IE per cent.
There war© times -whom liquefaction by th® above method
did not produce eaosistft&t results* ©specially when th© pH
of th© mash was lower than § and th® temperature of thinning
was greater than 70° C.
In this case, the mush did not
liquefy wsll and reasljidd as a thick semilltutiu
such a mash,
for scats* reason* refused to liquefy well even when additional
mold-bran m s added*
Th© resulting .alcohol yield# ware also
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
~36»
abnormally low.
A properly thinned mash always separated
Into two layers, a water-thin. supernatant liquid and a
sediment layer.
Such nasties eoBSlsteatly gar# very satis­
factory yields upon subsequent aleoholie fermentation*
f©
obtain such a sash the procedure employed was as follows!
The cooked cassava past#,, fresh from the autoclave, was
allowed to cool to §§0; 0,
At this temperature th® thinner,
in the fora of a suspension in water as described earlier,
was added and. the mixture was stirred*
place in a few minutes*
liquefaction took
After letting the aiasii stand for
fifteen to thirty minutes a separation of the layers started
to take place.
This method was found to give consistently
good results*
8* Temperatare for tJhlaalmu
A IS per cent cassava paste-
prepared with sufficient aeid to bring the pH t© § ant cooked
for thirty Minutes at 15 pounds pressors was found to be of
©
sufficient consistency at #5 C* to allow miring in of the
.
thinner*
More concentrated pastes were somewhat isor® diffi­
cult to handle.*
The pastes were more workable the higher th®
temperature but easymic activity decreased quit# rapidly at
these high temperatures*
Th® following experiment was under­
taken to determine th# upper limit of temperature for thinning,.
Thirty-six grams of cassava were gelatinised with 150 ml* of
water containing 3 si* of 2 normal sulfuric acid (to adjust pH
t© 5) and the sample was autoel&ved for thirty minutes at 15
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
pounds- steen pressure*
allowed to cool.
M
Tba flask containing tba a m p I® was
s m m as th# ianparature reached 85° 0.*.
10 si* of moM-torsta suspension aontalalag 0*18 grans moldbran was added and tba mixture stirred with a glass rod*
After fifteen minute* tba saftple was rapidly sealed to 30° 0*
by aaana of running tap water {14° 0.).
Bflployi&g bb@ gtag
procedure tba temperature of tba past®, at the instant of
adding the steld-bran suspension, was varied'fro* 80® 0* to
#8° 0,
Th# past# was oonaldarad wall thinned if a separation
into two layers w m obtained aftar two boors standing*
The
results ©f these arpariaants era shown ia Table XXXI*
.-
TA3SLX 1X11
Iffoot of fanpsratora on tba Thinning of Cassava last#.
Initial
t’aanaratora
faaparatura after
88°
83°
is •
80®
78*8°
IS
75°
?#
13
O'
fB
74®
80
»
0
70
69°
18
n
70°
6°. 53
m
n
®S°
«4.i°
.18
«
05®
§4°
30
Adding Thinner
Minutes
StandlniE
Degree of
Thiiminfs
unsatisfactory
«
satisfactory
H
It is seen from fabla XXXX that satisfactory thinning was
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
-58oht&l&od
mmn
©
at ?5 6*
This was th# upper limit ®f t *p i f-
ati re for tba notarial* aaployod aafi th# oaadltlos* of th#
fh# thinning or llfwsfaoMoa of the pastas
oxparlaeat*
saaoad to proeood m o m rapidly at tba ! § » ? toBporaturos*
Tfcia is probably du# to th® laaotiYatxon of m Tar®# part -of
tfe® ©naps# at th# higfear t m p m m t m m B *
for this roaaoa th*
lowest faaolhla taagwectura is «d#i«atlo is th# tbiaaiag
op*ratio&+
h. T iiim lm with baaboilal t M f w i *
Tim m M - b r a n
«Bploy*d is th# ozporlmoat* doseribod prwloualy ooabalood
o&aynos whioh war® aatlro at not blgfcor thus ? # 0*
*&
o&syao frcn Baelllua aaaantarioa* has boa*.kbowst to
ataroh siren at as high a toaporatur* as 95° 0*
sy*», knows W
y
Suoh an ®n-
th# trad® aaa® of "Sapidas* 10X”, was ob­
tained fmm th# Wall#wl#la Cottpaoy and its suitability for
mashing o*saarro was dotoanalaod la th# following manners
W
of a gram of th# osaym® la th# fo m of & powder.,,
was disaolwod la 20 ml* of distillod water*
A wolgbod aotouat
of g r o a M oassaba root was aixod with though distillod water
at 70° C* to sake 80 si. of mixture*
One si. of the "Sapidaso”
solution was addad b y -*#«&« of a plpott*.
heated on a water bath to BO® 0.
Th# mixtwro was
Th© ooasistoaoy of th# mix-
turo after ton mlautos hostlag was a©tod.
Th# results ar#
shown in fabio XTf.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
*■*00s
**
M l
Ilf
Liquefying Action of th# Bacterial lasyai# ”8apl&ase* on
Cassava Pastas
For Oeat
Mash
______
Crams Sample
Crams "Rapidasa"
Par 80 ml,
Par 100 grsa
Mixture______ Simple
Liqua-.
faction
15
11
0.908
If* 5
14
0*1ft
n
n
n
m
0.190
m
»
w
25
It
0*125.
slow, hut satis­
factory
SO
90
0.104
quite thick;
thinned slowly
it
.
very sat infant m y
Th# atev# results daoenstratn th# suitability of th# bacterial enzyme "Sapidasn* for thinning cassava fast#®*
fh#
advantage of this maxymm prefarsMes over asl4»bra& lias la
its ability to aot on th® oaasava starch at temperatures math,
high#!* than th# gelatinising tnnperatura*
ooaoantratad as SO
pm mn t
Cassava mash®# as
were obMnafela wh*n th® amount
of ansyan asad was only Q..184 par sant ©f th# weight of sample*
this Material nttsyn» U &w m mr k m m
aaoaharlfjrifig f#w#r*
Th# starch is transformed only Into dsxtrlns.
Mold-hras. is
therefor# still needed for th® formation of fermentable sugars*
However, this b&eterial snxyas is a very satisfactory solution
to tba nashlag problem presented toy tba thick, mucilaginous
Ohara#tar of concentrated cassava, pastas*
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-# 0 -
8*
8a©«J»rlfleiiti©ii by mold * Bran
a*
lash concentration, five hundred si* Srlenmeyer
!
flasks were ss&rkei. to contain z m ini* ■ Various amounts of
ground cassava root (10, 8©, 40f 50, arid §0 grams} corres­
ponding to 8, IE# 1ft,'St,.and 24 per cent ®aeb were weighed
in*
A quantity of-soli-braa eorrespoadlag to 1 per oeat of
the sample was added t© the flask and nixed with the sample
to act as, thinner*
Two hundred si* of distilled water,
heated t© 85° 0* were added with continued stirring by means
of a glass ret*
The pH was adjusted to a value of ft by the
addition of % aorisal sulfuric aeid*.
She mixture was cooked
for one hour at 80 pounds etdem pressure*
After cooking,
ssccharlfloation was all©wed to proceed at 60° C. for an hour,
with § per cent by weight ©f a©ld**bran*
added to auka .Eh© aCU
Distilled- water was
The sashes were Inoculated with 10 ml.
of yeast culture, incubated at 89® 9* for 72 h o w s , and dis­
tilled*
The results m m scows la Table XT.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
t £ M M Hf
'BtluuioX fl©M. from CtaUNftm tfftOfeoo of f e l o n s Concentrations
Bmp A m t
.
Mmh
Qamma
Samela/250 Ha,*
fsir Seat Si OS
8
89
88*8
12
9*
a$»ft
1#
4©
8**8
28
5©
88**
24
m
81*8
Glucose la Soapier ff..l Per feai
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
-SIfla above results indloat* that highest ethanol yields
Wfte obtained tvwk %$ p m
The slightly
seat easamvfi mash*
lower yields a t higher sash «<mo*a&rfttloii« itti perhaps dm®
to th© high eoaeeatratloii .of ethanol obtained.
films,, the 14
per ©tut stash after f # i » » t m t i © » s contained 20,96 g r a » &f
ikimlml p«* Si® Ml* of roash.
this, is equivalent to a eesoejitra-
ties of 8*14 pm? m m t ethanol witieli is about the H a l t for the
growth of yeast i®»«bll3J»tis®i to high alcohol ooneoatrotioa**
fb© higher o u t
were difficult to prepare due
to the stiffness of the paste ©y gel femei*.
W H & o u t very vigor­
ous stirring Imaps were likely to for® ant reaain unacted upon
by th# «apl*«o»
A rneh concentration of IS per cent seemed
to be about the upper llstt for this sethoi of mailing in
regard to ease of working.
**■ Sffeet of mold-bran ooneeatratlon on eleohol yields*
forty g r a » of ground « u c m root were sired with 0.8 gram
aoM^braa as thinner asi aashet with 8@0 si* of. distilled
water at S # 8*
''sulfuric meld*
fh© pi w%s adlusted to 8 by
snnuui
of 8 normal
fhe aistwro was autoolaved at S® pounds for
one hour and eocAoA,
Sac char ifi©at ion was ©tarried out at
§b® ®* for an lour wst&g various saeKnta of tuoM-briia.
fie
gnashes were inoeuleted with 1® ml* of yeast culture, diluted
to 880 si** l&eftfeKted at 80® 0* for ft lours, ant distilled,
fie data obtained, ere ilvwa. in fable X U *
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«
63~
tmm m
Sffeet of Meld-Sriiii Oosoostsrotloa on Bt o a o l Yields
Per Cent MoldBran Total
Per Cent Mold-Bran
for Saooimrifioation
Per Cent
B* 0H
2.5
0*5
64.0
5*0
3.0
71*4
7*5
5.5
78*5
10*0
S.,0
81*0
11,5
10.5
88*8
13*0
84*5
15*0
01uco.se is sample t
'
7**1 per cent
The above result# allow that, for the sample of moldbran e m p l o y e d t h ® hipest yields of ethanol w©r© obtained
with a total of 12*5 per m m t mold Bran*
Of this amount*
only 10.5 per o.ant actually acted as saccharifying agent
Because th® 1*5 per oast employed as a thinner was rendered
Inactive by 'subsequent cooking,
o.
Iffsot of varying the amount of thinner*
The amount
of solfl-bran employed as thinner flays «a Important part In
the washing. process for the fluidity of the resulting sash
depends on the amount of thinner used*
th® thinner*, however,
is later rendered inactive due to the subsequent cooking
process*
la the following' mpmeUmab the total weight of
mold-bran was kept constant at 1© per rent by weight of sample,
but the weight of isold-braa employed mm thinner was varied*
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
Forty grass
ot grovustd
easiwrya root wo*o KixoO with
weight of aold-braa as thinner
water at 8 # 9«
wtm
mi. suufeed with WO
m glnm
al« of
After the fS was adjusted to S the gdxtttro
at IS pomade
mtmwm
pressure for
mm
hour and
cooled. fii# mash wfs ooeohorifted site stifflei eat rs>ld~0*«ui
to. mate tli# total 1#
pm* seat*
iooooloted with 1© ml* of
yeast culture,. and distilled after loot&t felon for 70 hours
at m® 0.
The reialts ate given la table OTX*
f&B£I M U
Xffoot of Varying the /mount of Thinner on Ittianol Yields
from gsoMtvo Sashes Booofcsriflod with Mold-Bran
„_________ B a t l o $ Yield
^r^iSFi^r^riwEI^^Sr^i
t
Thinner/s&eelmri-ifer W®£%
. fter i It OH
1
1©
Ilf
73,5
i
10
ltd
80.3
s
m
3t?
80,3
4
40
2t3
70.4
s
90
111
79,9
t h m
data laOioote flat wifi tils method nufeifie, and
with th# material® «n$2of*&»
z
to S per oont
as
tMa*i#r give the highest yields ©f alcohol if Hie total weight
of »ol#«hran aaoooto to M
90*
gor.oest of the snaplo.
with 3
seat t t l w f , the sash is so fluid as to Is easily wort**
til#,
f&iiy# is therefore a Aletinot. advantage in inereas-
ing the thinner
twm
£ to 9 per ooot* for there Is
& gain
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
In th# workability of the mash without any accompanying decrease
In yield*
Since th* thinner Is not fully utilized for sac-
eharification, being rendered inactive by subsequent auto-
©lavlng, these result# Indicate that, for the materials here
employed, 7 per cent »©ld-bra» is sufficient to carry on seeehayifieation*
The data also show th# importance of thinning
the mash , for in the runs employing 1 per cent thinner, fully 9
per sent was available for sac-charification, but the alcohol
yield was lower than la a well thinned' mash in which only 7
per cent aold-brmn was present for sacoharifIcation*
It is,
therefore, to be anticipated that If the sash could be thinned
by some procedure ter the same extent as that produced by 3 per
cent mold-bran, th# awM-bran required for subsequent s&ocharifieatlon could be reduced to ? or 8 per cent*
It might
be pointed out, however, that it Is not so much the -amount of
thinner used, as the degree of thinning obtained, for as was
demonstrated by experiments i® b# described later, even 0.5
per cent mold-bran,' properly employed, produced- very satis­
factory sashes*
d.
length of bla# p£
Forty grams of
ground cassava root, aired with 0.8 gram mold-bran as
thinner, were mashed with 200 ml* of water at 85° 0*
was adjusted t© 5 with B normal sulfuric acid.
The pH
The mash thus
-obtained m s heated at 15- pounds sterna pressure- for an hour
and tfe«a cooled*
Saceharification was -carried out with 3.2
grams mold-bran at ©0® 6* for one -hour*
Inoculation was sad#
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
with 10 ml* yeast culture aai l&etibfttlea was at 30® ©, a#
astt«l.
4 pair na* distilled every 14 hours.
Table ZVXXX
presents results of these oxporiaoata*
"1$»YOftr
Effect on Ethanol Yield of Time of Fermentation of Cassava
Mash Saccharified with Kold-Braa
Time ©f Fermentation,
Per Seat It- 01
Hours
.0
0
24
44*3
48
it.T
?m
M S
«*
BOS
120
QisooM In samples
The data A m
§2.6 Per Cent
that beers should he distilled 'after 3
t© 4 days incubati©*.
m i s psrttoula* run was observed to be
rather liet, toot there was practically no change ia the yield
even after It# hours or 3 days,
ToanMBtod mashes have a
tendency to bee*® soar due to- @©ati«i.aeti©n with acetic
organisms.
Since these fermentations employing mold-bran are
not carried under strictly oooptlo conditions,, there is always
th# danger of aootlftootles of the oleehoCU
The beer ia this
experiment attained a concentration of 5*19 per seat ethanol,
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
•Hi?**
which Is low enough %& b© conducive to attack by acetic
orgeats®#.
#*
iffeet of Imeemliai yeti©*
forty grants of ground
mumwv* root were.sashed with A 00 si*, water at 80° C*
A weight
of laolS-braa ©qua! to two par #©»t of the ssspl* was asst as
t&inser*
lb© strtar© was amtsoiarst at IS p©«afts for 40
minutes*.
Upon cooling it m s saeebariflei. with 8 per oast
soi€-lraa at m m 0 * for «a* fcour, iaooulatai with yeast cul­
ture i» varying ea&ants, and diluted to £88 ml.
was at SO® 0 * for f& bears*
Incubation
lb© results are giy«a in fall# 12%.
Iff@ot of per 0g&t Inoculum on Ithaaol Yield
Inoculum
Per cent of
Total Volume
Per Cent Si OH
.I
f?*J8
4
fi.*l
M.
W
&{
% £%
3
w*
a
77.0
10
77.0
1&
ff.l
Th© abor© data-show that th# volume of laoeniwt# in th©
range studied, does not apyseeiably affect th* yields*
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
Th©
—Hi!**
highest yield «as obtain#! *, hmtmmw*
«sewr|: of .laomxXwi is «
mtmmxm
mb
# per Mat*
Tto
of th® masher of yeast #®lls
imtr©tu#®4*
*•
S
i
l
l l g M l s l a
atoftlMUtlflftfttKiftft# A sampl® of oftftium .root was pass®#
a tarr mill f e w times to
toother
ama&lm
i m m m m %i»
through
fiaemsiw of smMitisiom*
was ptssst last on*# a® usual*
Hashes wife
prepared fron these saaplfts usiag 36 grams la £50 mi* water
at 8 # 0.
fhl.iuil.iig was Harriet out at fl# 0* by the addition
of 10 ml, of jsolt-braii ftnspftasion prepared by stirring 1,8
grams
w&M^hrm
In 100 si* water at 40°.0*
mriisMes war# stmtlets
The following
fiaeness of Mapl®, stirring taring
thinning, and eooMng pressure.
SatoharifioatioH im m e«rri®A
out in all samples by neftas of 10 per sent by weight of moldbran at 60° 0# for 1 tow*
All were inoculated witfe SO si* of
yeast culttire imombatei at 50® 0, for fS tours*. The results
are stow in Tab!# 2X»-
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
« !§ .§ »
t ib ii a
Iffill os Ithanol Yield of Ftne-Orlnd 'lag,f Stirring «at
Pressure ©f Cooking of Cassava J-fash
Tlmmmm
Stirring
■Tiae*.
Mswtes
■ fine
#
Cooking
Per Cent
Eh OH
Pressure.,.
2©
90*9
0 '
78,6
90
©
80.9
«
#
it-
ff*l
H
If
20
ft,?
«
90
.It
ff.f
ieixii
©
n
Glucose, is s**$l*x
85*2 Per Cent
This investigation **» carrie®. **& f»*«a** th® slop er
residue after f o m o a t s i i i m ea&t*la** parti©1## #f th# original
Material urn**** upon by the noli mgrl***»'
A
atsrosoopls
«xaain*tl*& revealed plooss of the original root still re­
tained their ah«p* although the ■starch granules war® 'Swollen*.
It was thought that fine-grinding uni shirring m h m M eliMaate
these particles, for fine-grinding ifiiiwtM giro snail particles
that would he readily ***** upon by mold .wayles#* sit* stirring
should break say lamping that m y occur.
The pressure of cook-
lag say also be a factor is the disintegration of the particles.
Higher yields were elrteiite®. when the sample was ground
fin®., High cooking pressure w
of doubtful valuef the lower
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
pressure even showed an advantage*
this result la perhaps
explained os the basis that th© cassava starch granule is
small* gelatinise# at a lew besiperatur© and thus ”cooksw
easily 'without th© need of pressures above atmospheric.
The advantage of stirring was not demonstrated in a clear
cut nmn»er«
'fkmm was m
lagwevMMk&t la alcohol fields with,
stirring when the ooofclag pressure was 20 pounds.
But there
m s a slight gain with stirring when cooking was tone at
atmospheric- pressure.
«"* I*a*th of tlse in the, aaaehiuelfytflg hath
Thirty-
six grass of ground cassava root m s gelatinised with 250 ml.
of water betted to SO® C* ant amtoelaved at 20 pounds for an
hour.
After cooling to 74° 0. it was thinned with 10 ml. of
a 1*8 per cent suspension, of aoM^feraa and allowei t# attain
the temperature for saceh&rifioation by setting' the flasks
in the sacchmrifylhi hath which was held at 60° 0*
When this
temperature m s reached* 10 per cent fey weight of »oM'»feraa
wmm added and the mixture was well stirred fro® tin® to time*
■fb« samples were allowed to restaim in the bath for varying
length*, of time*
They were the®, removed and quickly cooled
under th© tap to room tanpereture*
The cooling required no
more than two minutes for the tap water was at IS® 8.
The
s«ples were Inoculated with 20 »1* yeast culture*, incubated
at 30° 8. for ?2 hour# and distilled.
The data obtained are
given In Table S I .
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
~ n ~
tmm
s i
Effect on Ethanol field of £aagft& of tl»# of Sa®ihiirlfl®«»
tion at 60° C, of Oassaire null
Saeoharifieation 3 tae#
Minutes
Fer ®mt
th OH
IS
m
m
?3*1
m
ft.S
■H» results mmm to iailost® that a saocbarlfi cat ion
him® as abort «0 li -Bloats* Is oooOnoito to felghfif yields*
The Im m eom'reraioaa at longer period# *oy fe® 4m® to the
growth of t:h«»©phi!I® tooterl* or leeetltstieo of th®.
SBxyse*
h.
tOHBsiistaas of seeobarlfyjflg hath#
As experiment
was eertied oat oslag the urnm set-hot aad 0*000$ of neterieXs
as la th® prees&lag experts®at* feat the ooooboriflootioik
teapsratmr® mm vsrleft*. fhe his® of seeebsrlfisetlea was
kept coast ant at on* tear./ Semite mm glow* la Table SIX.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«*Pin
/&«*
1M I M
txty
Effect of Temperature of Saeelinrlfyiiis' l&th on Xt&ft&el
Yield from Cassava Mashes
Temperature
®C«
.Per Cent .It 08
30
75.5
40
66*&
m
73.5
m
72.6
-
«hil» the fields in iMft experiment were rather low,
the data show- a
trials at various ttses
and &0° 0,
for higher field# at 30® 0.
wmm
further
ran. at the two temperatures, 30°
'The. results, as showa. la Tahle H i l l indicate
that the saeoharifieation process amy as well take place at
30° 0. as at 6 # 0.
TiffiH XXTIi
Comparison of Ethanol Yields from Cassava Mashes &mh>©harifiM at 30® 0. and at 60° 0,
Trial l«feer
30° 0.
80®' 0.
1
78.4
79.0
%
f«*5
ft.4
.3
0G*8
98*4
4
80*1
fS.t
%
84.2
85.5
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
The ia&m m m ub lu yield in the later trials* ©specially
the last, 1# ime to the thinmlng technique*
ftttnoastrato that,,
the
The above results
the ooaftitSoiw used In' this «xpori«oat»
aaoofeoYlfiofttioa procedure wherein, the Bash Is kepi at 6G°C«
Is & superfluous process «aft nay fee #liaiaat«t*
That a tempera­
ture off 80*> G« oorkn Just as well m §©# C. m y be explained
in the following Mk&B«rt
It is well km®m that eaecharificatiea
of starch by amylase toes-act g# to ®mplebica but reaches s
state of equilibria between the reactants ant the protects of
reaction.
feast ©amps®# act ©a the-protects,, setting up
another ronotion.
The process of couTOrsioa of starch to
id ©©tael in the pres w o n of nfflaco an! yeast consists therefor©,
of'two consecutire roaetioa* ititch say b® fonmlntod tteist
te&cbiea 1
Starch
Seaetioa ft
ifeXtoao
-p .%©
4- % § Tenet
Httt©#®
v CgHgOH +• 00g
The first reaction proceeds forward ant approaches completion
because the prefect of reaction, salt#®©, is removed through
the second reaction.
If the ooeoaft reaction is slow enough,
that is, slower than the first reaction «t 30° C. then there
M i l ' b e no difference in the alcoholic fermentation whether
saccharifieatioa fBe&etioa 15 take* place at 60° 0. or 30° ©*
According to the above experisettt, the role of the
;thisner is to mmm& the starch into soluble ferae, oooBKraly
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
designated as dextriixs.
The literature retards this conversion
to he optima at About 50° 0* snfi this is found t© be the case*
But the results obtained in this oxporfjMst tend to show that
the conversion of the dextrine to salt©#© is better achieved
at 30° G* rather tten 60° 0.' fbe- higher ts^sratmr© for such
a prolonged tin* as 1 hoor'auy iaae-tiwithe the m M
tasya***
Sinee there is reason b© mm.feet that inactivation at the 60° c*
bath is conditioned by the length of tins the simple is heft at
this temperature* this factor was farther studied.
1*
interrelation of mold-bran concentration, length of
iisa la & • sail. M S . sai z M S . si sStessl-
sarty-six
grans of .ground cassava root were gelatinised with 2S0 al.
of water, at 80° 0*t eontaloiiag 3 ml* ©f I normal sulfuric
acid ft©- adjust, the pi t© ■§)* fweaty^four such simples were
prepared in 500 si* flash© and heated at 15 pounds steam
pressure f©r an hour*
the- samples were allowed to cool to
§-# 0 * fen ml, of a. 1*8 per -cent niold**bmo. suspension were
admixed as- tMftner#, and the mixture was- -allowed to stand for
15 minutes with frequent atehiag*
three concentrations of
a*ld«»hr*a were staled for saccharificatloni
per cent*
5, 7.5, and 10
the lengths of t i » la the saccharifying bath were
0, 18,. 60, and 120 Minutes*
In the css® of the *0” minutes
me«fea«if£efttlo& the fleshs were first cooled to 3Q° 0,* and
then ssold^brsji. m s added*
All the flashs were inoculated
with yeast and incubated as in previous experiments*
the
results- are given in -fable XEE?»
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
fJ p T .1 l Y T T Y
Effect on Ethanol Yield of Varying the Mold-Bran Concen­
tration and the Time of Saceharification at 60° C. of
Cassava Mashes,
*
ifeM-lran
Cone*
l
'i
length of S m © in the 60° 8* Bath,
. . Matties
-
-I
nor—
“ i i r “"—
5*0
78.3
?i*4
76.4
75.6
y.,s
61*4
64.0
80*5
80*8
10*0
04*0
§4*5
83.5
80*4
B w results obtained in this experiment indicate a
elicit advantage of carrying ©mb saccharifieation for just 15
■lushes at 60° 8* -With these eentltlea* of tin* and tempera­
ture it is found that f*5 per cent &o14U1»**b was sufficient for
obtftiaing good yields of ethanol-*. The data given, la Table K I T
show that alcohol yields are affected adversely when seaoeharification was allowed to- ftoceei at § # 6*. for one or
two hours-.* la fact better results were obtained when the
saccharifying or "malting* hath m m entirely eliainatet and
the mold-bran introduced into the cassava sash at 30@ 0*
Xros the shove ye-suita it was recognised that, proper
conditions for saeejtairIfieation have m decided effect on yield,
for this reason a study was carefully n A « of the process*
Various' tooporaturoa of the saccharifyisg hath and different
lengths of bis® In the hath m m
Investigated*
The method of
preparing and thinning th® staples was -as la the proceeding
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
©3qperiii«it*
'Tkm proportlon
of isolA-hram eopleyed was ?*S
per eent of tins weight of the saaple*
war# aats: In the following manner:
sample, tli# repairs! aacnait of
She m a s ■at 30® €«
fo the prepared tfeianad
woi/Mvemn was
added and thm
flask was allow#! to'rao&ln at Si® §• for dd aUnxtes is one
©ass aai 12© adnatas tii the other feafore inoeala&iea with
yeast was easarfed <wt*
411 runs war* la doplieet* as nsnal*
Bxe results are presented la fall# M T .
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«*??■*
T-Afnyn s y
Effect on Ethanol Yield, of Temperature and length of Time
In the Saccharifying Bath
Temperature
of Saccharify­
ing Bath
Time in the Saccharify­
ing Bath, Minutes
Per Cent
It OH
.90
84*?
m
84.5
40®
15
84.4
40°
m
84.0
40°
-so
83.5
40®
im
81*8
§5°
.15
99.8
SB®
30
83.8
55®
50
81.3
55°
190
60°
15
S5.4
60°
50
80.0
mm
50
78.3
190
73.8
90®
so®
.
i
80.i
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
w98*»*
The jrftftult* obtained from tilts eoqpsrlnuot as presented
la Table H I , demonstrate ©nc# nacre that the saccharifying
#r ^nftltiag* bath, la' m%. ©sly unnecessary but M s a tendency
to lower yields,
for this reason it may be eliminated, thus
eia^lifylag the alcoholic fermentation of starchy Hashes by
the allslaetleft of one eparatlaa*
the fniuLtiag* operation
M s always been consider^ very essential la. the' brewing end
alcoholic femeatattea proses®## estpieylng salt for ©©aversion
of starch to sugars*
According. to faeofea end Menton (1938),
one objection to the ms# of salt aa saooMrlfyiag agent is
the ooiimertial production of ethanol is the tin® and careful
control refulr-ed to- hold the eoofceft starchy mash at m certain
temperature to Insure .complete iiastati© action,
Frost the
results obtained In the present experiment, it seem® that this
objection could Ira removed by the replacement of malt by mold**'
bras*
With the employment of aold^braa, the saccharifying
or ^talting*1 bath may bo allmihatad*
introduced into lit natch at 30° $*
The sold-brsn is simply
The t«a##r«tura. Is not
critical* for mm demonstrated by the debit In fable X U , there
is 90 practical.' differesce is final ethanol yields whether
the aold-br«n, is added at SO® "0* or 40® 8.
j*
Saocharlflcation at temperatures above 60° 0 .
^ploying a procedure essentially identical with that used is.
'the proceeding experiment higher saccharifying temperatures
were investigated t
65°, 70®, ?§®» -ant 80®.
The thinning, was
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
started at i # 0* mxA saccharifioatlon m,* carried out at
th* d#»lr@fi temperature.
mad# at 60®*
For comparison, ran# w m m also
Saccharifiestion at 60°, 65°, t M f # w@r©
carried for an hour*
Sacchariflcation at 65° was esrrisd
©at for 1 minute and § Msilii*
i»i only for 1 alnat**
Saoeharification at 80°
the results ay# given in fable XCVX*
yOT'
fIy
XXf
c t on Ethanol Yield of teployiag fsaspcraturiis Above
0» for Sacchariflcation of Cassava Hash
T«meratere
Minutes
Per Cent It OH
60
m
fi.lt
is
to
®4*f
to
to •
57.5
fi
f§
' 80
■ i
S6.fi
3
i f *4
1
if.lt
.
Til© above tats demonstrate that tissperatuyea: above 60° 0.
give low yields ©f alcohol.
These low remits m y be ascribed
to destruction or inactivation of the ® @ M ensyaes.
**
H U g , gt m M l & m MMmm&.jmmSii*
The materials
and methods in this experiment war® the same as in previous
runs*
The improved method #f thinning at 65° with 10 'ml* of
1*8 per cent »oM*toran. suspension for 1 hour and saccharifying
with 7*5 per cent .sold-bran at 3§# was employed.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«
90*
fte results «r* shown lirYahl® XXVXX*
.
tAB£X M l
Iffeet 'on Ethanol Yield, of Employing Yayi©ita Yeasts la
th« Fermentation of Cass&m Hash
Fey Cent St OH ..
.... Yeast M M r
2
78 #0
14
80.2
21
82.2
98
.80.8
m
84.0
43
83.5
Yh© data stetatasd indicate yeasts SiaOmF 81* 42 and 43
were found test suited to th© ©wuMrnt smhstimt® used la this
iaYestigatioa*
Hsrib«rs 42. « i 43
-mm
slightly hstisi* than
laatosj? 21.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
—
ox*.
4*
SaseMri fi cation with barley aialt.
«.
Effect on ethanol yleiaa of varying the malt
pentrat Ion,
bob-
Thirty grams of g-ttss®rfa flour was gelatinized
with £30 ml* of water containing 8*5 si* of 8 normal sulfuric
acid soft heated for 4© minutes at IS pounds m % m m pressure*
After cooling* sacchariflcation was carried out at 60® S. for
an hour#
Various proportions of hurley malt were used*
the
sashes were ittoeulated with 20 m 1* of yeast culture and al­
cohol was determined after ineubaties at 30° 0* for fi hoars*
Two runs were ..saute on different days, each run being- in
duplicate as usual*
The results are shows- in Table XXVIII.
m m M xxvxii
Ethanol Yield from Cassava Mash Saccharified with Various
Concentrations of Barley Kali
Per feat it OS
. Malt Coaoeatration'
.1
8
l*f
4i.S
4t.t
1
59.6
55.0
10
68.4
62.3
80
69.5
ti.f
-Cora with 30
per eea-t self
59.9
i®.i
The above data show that -ethanol yields increased slowly
with X m m m t m proportion of salt in the- mash,
The yields
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«68*»
were rather lew ©fen
the mgna% of malt used was 20
par seat of the m m m v r n sample*
*>•
98 etjLanoX ylald of TOrylng the cassava aaah
concentration.
late flasks, market to contain 250 ml. of
Hash, 20 grans of ground « i « » « mad 0.2 gran® of malt {1
per' seat of sample} we re introduced a n t itaishad with sufficient
water at 8 # 0. to make up a volume of ISO sal*
s&oaer, pashes esatalAlag 30
la the sag*
grants!f 40 g e m m mat SO grants
cassava per £S0 ml* of mash were prepared.
All the gaaples
were allowed to stead for 2 hours to complete the preliminary
The sashes were- then amtoelavei at IS pounds for
lifuefmotion*
40 minutes*
After cooling, sacchariflcation was serried out
at #0® S, for «a hour espieyiiig a quantity ©f smalt equal to 9
per sent of the sample*
The inoculation with yeast and in­
cubation at 30®' G« were carried out a® usual*
The results
are given in Table S I X *
TABU rrrv
M t m m l Tleli fro® CtaMMnrm Mufti o f furious Concentrations
Saccharified with Barley Halt.
Mash Concentration
Ground Cassava
oar 250 ml. Mash
Per Cent
8
20
65.6
12
30
70.5
li
40
74.0
IKf
90
73.0
Per Cent
St OH
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
The data show that cassava. nashe* prepared with this
m b
pi® of hurley salt gave almast identical ethanol fields
when the sash concentration was IS., 1ft, or 30 per sent.
The
y l f M s wars law indicating that malt toes not ant well ©a,
M S M T S
«*
»§t®4l*
Hffeot of varying the properties of thinner to
aaeoharifler on alcohol yields.
The procedure employed is
this «p#ri»@at Is identical with, the proceeding*
The amount
mf salt as thinner, however, was varied. so that the thinner
constituted 0, 1, ft, S, 4, ani § per test of the sample,
The
corresponding -amounts of malt employed for sacchariflcation
were
1 0 f
t ,
8*.
6 ,
and
§
per cent of the sample.
words,., the total «osst' of malt was held
Is other
at 10 per
cent of assigl®.* the results m m given is Tahle S I *
TJBUS v w
Sthanol Yield from. Cassava Mash When the Proportion
Thinner:Saccharifier was Varied; Barley Malt Used.
Thinner
Per Seai t’ 'Far Wia€ o£
©f Sample ; Total Malt
0
§
;
Eatio
s thinner jSaccharifl©r t Per Cent
t It OH
..».......
•ft
•
*
0:10
67.2
.1
1©
1:9
07*0
ft
.SO-
1*4
64.6
S
li®'
Slf
07*2
4
4®
2:3
66.*
5
SO
1:1
07*5
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
fto© data show'that* varylhg the proportioa of thinner
to saccharifier lias no apparent effect upon alcohol yield.
With mold-bran there was a definite gain in. yield when the
ratio of thiaaey to saeol»rifyiag agent was ltd or f?7.
All the sashes la the present experiment were fait# thin
■ant fluid*
the apparent inability #f barley salt to giro
higher ethanol yield any he A m to the nature of the «sty-
las® or amylases present in emit*
the differeaee la
behavior hotwoen iseM-braa ant unit in this typo of ex­
periment lends support to this view.
d*
CteaHMuPieea of aoit^hrea and iwrlnr gelt as nan-
charifying agents In the mleeholl# fefgentstlon of nananta
gash.
fo 40 grams of ground cassava* 0*4 gram of seld-braa
or barley salt was added as thinner and the nixtnra was
Bashed with £00 el* of water at 8S# 0%
The samples were
then heated for an hour at J8® pounds s tow pressure.
Sac­
charin cation was carried out at the i # hath for m, hour;
isold-braa ©r Parley -salt m s employed depending upon which
aaterlal was used as thinner*
In nan experiment the samples
were thinned with malt ant saccharified with aold-hran.
usual' all the samples were laoenlntnd with yeast sad in­
cubated for 72 hours at- 80° 0.
The ethanol yields are
presented in fable S S , ■
•
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
An
TABLE m i
Comparison of Mold-Bran and Barley Malt as Saccharifying
Agents in the Alcoholic Fermentation of Cassava Mash.
Saccharifying
Agent
thinner
Per Gent
It OH
1 Per seat Meld-Bvaa.
66*8
Barley Malt
1 Per Oeab 'Barley Malt
fS*«
larley Halt
1 for Seat Hcld-lraa,
fS'.t
la *his experiment the effectiveness of salt as m liquefy­
ing and mm a saocharlfyiag agent m s i«onstr«t#i separately*
The ethanol yield m s low 1st the mash thinned and saccharified
with wilt*.
The yield increased when, the thinning was carried
out by m m m of mold-bran instead of malt.
It way he con­
cluded that salt amylase is rather deficient ia the
which liquefies starch*
It wag observed, however that the
wmsbm which were thinned with malt were as fluid as those
tfelaned with mold-braa.
A possible explanation to these
rather ^atradietory results may be offeredi
The amylase
la malt differs la nature fro® that la iaeM-braa -sad the
difference lies mainly la the sbereh^iifeefyiag. component.
It is therefor® probable that the primary comrereion pro­
duct® txcm starch are not Meatleel,*
The thinning action
is them aot only physical but els© chemical*.
The products
formed wtmm nold-hsia is «pl@yed to liquefy cassava starch*
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«•&&*>
are- of swell a nature as to be readily aefced upon further
by the tmflAtm in both malt or janM-bran*
When salt is
used ms the thinner thm prisary trsnsfow.tioa pro&uete
are probably m & f resistant te feather' nobles of the
enayae in salt*
Whatever the m m ® § the superiority ©f a»X&*bra& over
salt, as seeeharifylng agent in the aleoheli© feraentabiom
of eassara stash tet been demonstrated by this experlsMat.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
-87-
8*
1.
Butyl-Acetoni c Fermentation
Replacement of corn by eassaro*
Three hundred ml* stashes were prepared In which the corn
was progressively replaced by ground cassava*
The weights
of corn and of cassava are given in Table XXXII* The.pro­
cedure was as followst The weighed quantities of corn and
cassava were placed in 500 si* erlenmeyer flasks and water
o
at 70 C. added and thoroughly mixed* The mixture was made
to gelatinize by heating*
The samples were then autoclaved
at 80 pounds for 40 minutes* allowed to cool
and inoculated
with 10 ml* of a culture of the butyl organism la corn mash*
The flasks were incubated at 37*5° 0* for 78 hours and then
total solvents determined by distillation*
The results are
given in Table XXXXX*
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
t m m ran
Yield of Total Solvents in the Butyl-Acetonic Fermenta­
tion of Corn Mash in Which 0 to ICO per cent of the Corn
was Progressively Replaced by Ground Cassava
Per Cant
Cora
100
Grams
Com
Grams
Cassava
Warns
Grams Solvents per 100
Total
grams of Glucose in
Solvents . Seasnio
18.0
0.0
4.16
30.88
m.
m
14,4
3.6
5.95
88*90
10*8
f.5S
4.50
31*9©
40
f.S
10,8
4.61
11.98
m
3,i
14,4
4,44 -
lo .a
0.0
18.0
0*.f4
0
Glucose in Corns
.
5,00
76.5 Per eeatf la Cassavat 88,8 Per cent
Table XXHI shows that butyl-acetonio f*»«atatlon tit
not take flaee to may large extent in a aash containing only
cassava,
Bewevar, a ea c m v * a u t containing SC per cent corn
Yemen*# satisfactorily with yield# of total solvents com­
parable to those obtain®*, fro® a mash containing corn only*
Cassava sashes containing 0 t@ 30 per cent c o m were
further studied.
Baploytng the same procedure, as la the
proceeding experiment and using a sample weight of '48 grams
to make 800 ml. mash, the experiment m s repeated.
The replace­
ment of corn in the wash was made in steps of 5 per cent.
results are given in Table XXXIII.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
The
-89TABLE XECtll
H e ld of Total Solvents from the Butyl-Acetonlo fermenta­
tion- of Corn Mash In "'hich Corn was Replaced by Cassava
Per Cent
Corn
Gram#
Corn
Grams
Grams
Cassava
Total
Solvents
.30
14.-4
m
12*2
m
Grams Solvents per 10©
Grams of Glucose in
Samel®
33*4
11.1#
2-9*0
35* &
11.03
28.#
9.6
38*4
10*44
87*4
15
7-*.,!-
■40*8
6.45
88*4
10
4.8
43.2
6*44
,17*0
5
2*4
45*4
3. 48
10.1
0
0.0
48.0
0*74
1.4
100 ■
21.0
0.0'
4*4.8
31.7
dittoes* in Gormi
.
74*0 Per east; in Cassava:
88*6 Per cent
Table XXXIII shows that la order to obtain good yields
of total solvents In the fenhyl-aaetoala fermentation, of
oassava, at least SO- per ©eat of the mash -should ha corn,
A d s fls ls fis y of cassava in protein or nutrients any explain
its low fsrmsnt&feillty.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
2* m s m s t th#
mi^mm figur,
sans* m
of shrimp mw&m* m m gluten a#ml«
isigi, s*s%am* M
m ® zisMsi ia£& J R & W & *
w s
M
s m s m
To SI graft of ground
oasasva* 150 si* of water were added and tborwiglily mixed*
Tbs pH m s adjusts#- to 6.5 by the addition of 0.5 ml, of 1
noma! sulftsris ncM*
.the storing* powder,
or^ soybean flour was then added*
mm
gluten ssiiX.,
Tit#■soybean ws* best intro­
duced by shaking the weighed quantity with a w ail amount of
m t s r la a test tube asft pouring tbs xuraltlug smpe-nsloa into
the mixture ©f mmum®. and water.
SaytXl ssKra&ts of soybean left
la the test tube were rinsed out Into the flask by means of a
wash bottle* .In the ease of peptone, urea, and expressed
yeast solutions or suspensions of known concentration were
made mad the desired quantity was pipetted off into the
cassava flask,
for exaapls,
$grans of peptone'was
dis­
solved in water to m i s 15® si* of solution and I ml. were
used for introducing 0.1 gram of peptone into a cassava mash*
The mixtures were heated under s t e m pressure for 4® minutes
at IS pounds*
Upon cooling the aastoes were inoeulats# with 10
si* of a eaXtwre of the butyl orgftatSK*
The total solvents warm
detealned after incubation for ft hours at if.5©- <j. in these
«p«rlaents ■the- sample of cassava employed had a glucose
equivalent ©f 8£*.8 per cert.
The correction for inoculum
as detssaiiied by actual fermentation was 0.139 gran total
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
-*Lsolvents per X§ ml* of ifioetiUou
The oth«r corrections, ia
grass sol-rents per «ra» material, were?
for siirlsp, 0ȣ6f
for peptone* 0*4S; for soybean, cexpressed yeast, oorn
glttfcea seal, and area, the correction was found to be neg­
ligible.
The reanlta of this erperiseaf or# preseatet la
Tables XXX1T, X O T , XXXVI, XXXfll, 2OTX1X,. and X T O X .
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
w § ll*
TABLE XBEFt
Yield of Total Solvents In the Butyl-Aoetonio Fermentation
of Cassava Mash Containing Various Amounts of Shriaap Powder
Per Cent
Grams
Shrlm®
*O vQLJL
Solvents
Grams Solvents per
..10© pernem, Giuoese
0.0
0.0
0.76
l.t
0.5
0.1
1.67
'»*«
1.0
0.2
2.38
U.5T
2*0
0.4
3.85
ti.i
1.0:
©.#
4.8©
Bf.i
4.0
0.8
5*26
3©*2
5.0
1.0
4«f§
tf.i
•
4 .92
31.?
'Cora
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
«*$$*»
TABLE H M
fi #M ©f Total solvents In the Butyl-Aeetoni e Fermentation
mt Cassava Mash Containing Various Amounts of Corn Gluten
Seal
Grams Corn
Gluten Meal
Grams Total
Solvents
0,0
0.0
0*94
1*9
0.5
0.1
0*09
3*6
1.0
0**
0*69
'5*9
2.0
0*4
2.28
13.1
3.32
19*1
Per Cent
Corn Gluten
Meal
3.0
Grams Solvents per
100 grams Glucose
4*0
0.6
4.01
23.1
5.0
1*0
5.07
59*2
4*9#
.31*9
Corn
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
<*94**
TABLE X X M I
Yield of Total Solvents in th© Butyl-Aeetonic Fermentation
of Cassava Stash Containing Various Amounts of Soybean Flour
Per Cent ■ Gnawi Soybean
Soybean
Grams Total
Solvents
Grams Solvents per
100 arums Gluooae
0,0
0.0
0.26
1.6
0,1
0,1
0.43
2.6
1*0
0*£
1.13
6*6
.1.0
0*4
l.fS
11.6
3.0
0.6
3.1?
18.2
4.0
0.8
M O
21*2
1.0
1.0
4.43
£6*6
m*
4.36
28.1
Corn.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
#*§§**
fiBIS ZZXfZX
Yield of Total Solvents in tit® Butyl-Ac ©tonic Fomentation
of Cassava 14a.sk Containing Yariosts Amounts of Owpreesei.
Yeast
Per Cent
Yeast
Ml. of 10
Per Cent
Grams Total
Solvents
Grams Total Solvents
per 100 grams Glucose
SasnoMioft
0*0
0.®
0.14 '
1.5
0.5
0*«
0*85
8*5
1 *0
i*a
0*40
4*8
8*0
8*4
0*f0
0.0
8.0
0.70
7.0
4*0
4*8
1.26
18.8
8*0
0.0
1.61
16*8
Corn
-rnm
.fweigkt of ausjAiii
86.6
IB gjNum}
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
maws m v m
Yield of Total Solvents in the Butyl-Acetonic Fermentation
of Cassava Mash Containing Various Amounts of Peptone
Per Cent
Peptone
JCU of 5 Per Cent
Peptone Solution
Grams Total
Solvents
Grams Total Sol­
vents per 100
m u m * Glueose
cut
§
0*§f
3*#
€UJt§
1
©*m
5.2
2
1*75
10.0
1.0
4
2.55
14,7
1.5
6
3.If
SS.t
2.0
8
3.3#
IS *5
4.0#
S3-.5
5.6#
23*2
2*3
Corn
m
•
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
-§f.
Yield of Total Solvents in the Butyl-Aeetonic Fermentation
of Cassava Mash Containing Various Amounts of Urea
for Cent
Ml. of 10 Per Cent
Vgtmo Total
Urea
Urea Solution
Solvents
Oraas Total Sol*
vents per 100
grams Glucose
0,0
0
0*88
3.9
0.5
1
0.85
4.9
1.0
8
1*06
6.1
8*0
4 ■
1*18
8*8
8*0
ft
1.16
6*6
4*0
8
1.36
7*8
8.0
10'
i.it
0*7
«*
8*88
88*6
Corn
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
Tables 2 HZf , fflf, XHII, XXXfH, XXXVXXX, and I S H
present the results ©f attempts to locate substances tfiieh may
serve as nutrients when added t© eeaae** mash,
M
repeatedly
sliows is these agparlaaat* eaaaeva by ibseM is a very poor
substrate for the butyl-aeetexils fermentationj the yields of
total #©Ivents were always very low*
The addition of small
quantities of shrimp powder, sorm gluten Jamal» aai soybean
flour was found to increase the amount of total solvents to
values ooMparsble to those t * m aosm*
Corn w
■used as the
reference material baaaaaa it 1# k no w to be one of the best
substrates for tli® tetyl-aeeboai# fermentation*
Shrimp powder
as far as known has not been need previously in an investiga­
tion of this kind*
Its effectiveness in promoting fermenta­
tion m a y b e asbribei to It# highly proteinous character. It
is abundant is m m ® localities like the Philippines and this
fact would favor its employment as a nutrient la the eon▼eisioa of essbatm into butanol, moetoae and ethanol.
Compressed yeast ant urea la the quantities studied were
not found suitable as nutrients.
Peptone in email amounts
served as an exasllaat nutrient.
Cm a large scale fermentation
heifsvsr cost is a drawback to its «pl©yseat.
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
m m u x e s m conclusions
1*
A study ©f tli# fermentative utilization of cassava
was undertaken*
Both the alcoholic and the butyl-ac©tonic
feamtftilea process## were applied t# ©assay#*
Acid* sold*
l«sa» and salt were Investigated as gacoharifylng agents*
S*
la studio# m
thm noli hydrolysis ©f very concen­
trated im#h#s* in nfhleh th# proportion of sample to acid was
carried a# high as lslt it was found that conversion of
starch to sugars was jOAxiam whoa the rati# of sample to
acid was 1*2*5 to 1*5 and when the concentration of the
sulfuric sold m # 0*4 noriaaL.* Yeast fermentation of the acid
hy&rolysates guv# poor yield# of ethanolf the addition of
mold-bran .improved the yt«M# *©Mn&at«
5*
Wien molcl-bran |la «oafun#tl#a with acid ) was em­
ployed for saccharifi cation of -pasles* cooking pressure ant
aoid concentration m m ® found to ter® llttl# offset ©a the
final alcohol yield*
4*
Several methods of thinning cassava a**h war® studied
bsowaa the usual method of latedmelag the mold-bran or salt
into the sash at 55°~60® C. was found to be difficult.
The
difficulty lay la the mucilaginous ©Mracber of «tii«nt paste*
Satisfactory thinning. was accomplished by stirring the past#
■with a suspeasioa of aold-braa la water*
The pro©### was
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
—1 0 © —
carried out between
m°
temperatures the m m k h
easy mixing*
and ?§® S. (best at 65° C*} at' whtcb
paste m i sufficiently fluid for
A toetarisl sassy®#
k&mm
commercially
waft fdMBl t# be a satisfactory thinning agent*
m
uHapidaaen
Mashes which
tod been thinned by means of mold-bran gave consistently high
■ethanol yields upon saccharification asfi fttntotttotleiu
la the
thinning protons the chemical change brought about by aol€»brsa
is believed t@ differ fires ttot brought about by amlt boeums#*
/
although both of.these »yloiyti# jMbsrlals were able- to liquefy
cassava paste to the Mam. exteat, the ethanol yields varied
aceeydtag to which mterial was employed ms-saccharifying agent*
8.
Yields well above ©0 per cert of the theoretical were
obtained in the alcoholic fermentation of m m n
under labora­
tory conditions, fhe procedure which gave consistently good
yields is
m
followst A cassava paste is prepared containing
M gmm* of cassava per 100 si* of water*
th* paste Is
liquefied at 85° C* by means of a suspension of xaold-br&n in
water | the «©-uat of mold-bran is 0*8 per meat of the weight
Of the sample. A quantity of mold-bran equal to- 7*5 par meat
of the weight of the cassava, is introduced into the thinned
mash at S#' te 40° .§* fto « s b is inoculmted'wi'th yeast and
distilled after 7E hours.
6* 'the .Ataaftsfd sftectorificmtioa operatioa- for otherstarchy mashes, known
for cassava*
am
halting", was found to to usneoessary
A study «ti made of the effect on «th®u»A yields
R eproduced with perm ission o f the copyright owner. F urther reproduction prohibited w itho ut perm ission.
-101*
of th® length of time, th® teapot®turn, a M the coneefitratlon
of mold-bran for saccharif ication.
fh@ higher the ten&eretur*
and th® longer- th® time for- sacoharifleatlon the lower «***
th® ethanol yields obtained.
ffe® best results war# nttataod
when th® mold-bran was introduced into th® mash at 30° 0.
M®
t«®feratur® 'Control was necessary b#om«s® yield* were almost
identical whether th® operation was. carried, oat at 30° 0« or
at. 4 # e*
f * 9he saccharificatloa of ******* sash with barley salt
was found to be e&Matlsfaefcoacy*
low.
fit® fi isl ethanol yield® were
Mold-brsn was found to be much more effective than
barley &alt.
8*
fh# butyl-acetoai# fexmeatetlo* of cassava by Itself
m s found to giro very low yields of total solvent®.
The
replacement by corn, of #0' per cent of th# w$i&fc of the
cassava resulted is yields *e*p«r*hl* to those from corn,
alone.
9b* effect of th® addition of various amount* of
shrimp powiev, « m
.gluten meel* soybean flour* compressed
yeast, peptone, ant urea on the yield of total solvents m s
Investigated*
Of these, shrimp ponder was found particularly
'fKrcedeiag because of its ready availability -in th® Philippine*
and became® it gave satisfactory yields of solvents when it
me* employed la small qttftntltiee*
f* for th® alcoholic and the- bmtyl-acctonic ferrasntations, it tree found that th® eeaeeve root aoei not b® ponied
as is ton® customarily*
9he pooling pro®*** estell* espouse
Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
-Mi-*
«il m n a m m tii« lass of fomoatftbl* material*
root nimm-
Tim unpeeXM
m m fount to im, as. suitable for fsrsanba*.
as tli# H our proporoA from. poolat roots*
Reproduced with permission of the copyright owner. Further reproduction prohibited w ithout permission.
jyOSSRASJlHS CITED
M w M m * f* T.
Th© m m & m isAusirj im the Fhllipptij©#*
Philippine I* Agr* 4, 271-285.
1933.
Adriano, f, T*, lames, s« T*, sad fn&lyess, 1. A*
The
pvoa&nate 9&«nlo*l analysis of Philippine foods and.,
feeding staffs?
530-34*
A r c h b o M , 0.
Ill*
Tim Philippine Agriculturist 10
1932*
€a*funra m
a soaipetiior of a»l2# in the
production of starch « t allied products*
.Ohiaa* Xa&. £2, 63-06.
I* So#*
1903.
Association of Official Agricultural <3b«nlsts*. Official
and tentatiw aetlnds of maalysis, 4th «i*
Bacon, S* F#
Itii.
Starch production la the Philippines*
Philippine 1* science.
JiA^. i3-i8*
1908.
Baefcfe&oo* 0* ¥1*. The use of ■various mild amylases in the
»a##harifi#ation of corn mmmh for ethanol femeatetioa
Onptthliehei. fhesis*
1ova*
Soiiifi, A.
tihmry, 1m m State College, Asm*
1940.
the Amyl# proeess.
Inc. lag* (Bum* JH# 712-13*
1933*
Christensen, 1* If* and Fulmer, 1* X*
Analysis of g-feutaxuiX
aeetoae, eat ethanol in aqtamme aolutios*
<tom»
Anal.
li* 7, 180-2.
lad* 'Sag*
m
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-104Collens, A« I.
Alcohol fro® cassava,
and 'fohsip, Bull#- i±> 56,
1914.
Copeland, S. 1,. H u l o c or cassava*
l#Vlew
139*188*
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Philippine Agfi®,
1908*
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0©11«®§ of Agriculture, ■
University of the Philippines,
Privet® eoaimmi cation
Becesiher 3, 1938 *
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ift-s©,
lot* sur 1® fiucoss,
Ana, Chlm, Fhys. El,
is4?.
! ! § » » S* B.
Ctam!®*! studies- ©» cassava products,
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Ewell, 1* 1* and Wiley,
mmi@m
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Soae products of ©assava*
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furao-ek, 1* 1* W.
1936.
885*89*
1893.
Poisonous constituents of sweet cassava
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1937,
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Gall®, i*
1931.
Bus Amyloverfahren unt seine AnwendungssSglich-
h®it@»..
Harada, T,
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1913*
Preparation of A* orvzae enzymes.
Cheat. B3. 14E4-37.
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1931.
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American loaraal of Pharmacy tf,, 988*58*
1914 »
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tsorg® Mica oat Unwin. Ltd.,
malting uni brewing.
Loadon*
Biochemistry applied to
198? •
la#©!** f * »* and Iiitea, S* f * Ifebor fuels. from farm products
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lilt.
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Die ▼®r«h«lbuag yam Iftaioka test -Spirltnt uni. Hefe.
leit* fir Spirituslndustrle 33* S00-201.
not seen*
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Original
Abstracted, in I* See* Qb&m. lad* 28, 618*
ItOf.
Leitemaa* L*
fatty acids
with cassava starch.
lour* Mm** (Mmt* So#*
H&reker*. M*
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Standpuakt unserer Kentniss tber -tie diastatischen
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Ha turf. Versamol. Munchen. tMS*
Original not seen*
559-60.
iStf-30*
16??»
Abstracted in ® M m * Zentr * JJ*
!«?©.,
Seadiola* I, B.
Cassava growing « € etsiafi.stareh manufacture.
Philippine Agr* jg|* 447-76.
llonier-Williams, S. W*
utilisation*
« *
foiwr alcohol, Its production and
Henry Frwsde
and Hodder & stroughton, London
1923.
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•106*
®mmwm
Moor#, 0. U*
starch*
its -oanfcant of hydrocyanic acid and
¥• .8* Bur, of O m * Bull* 106.
Moor#* 0* 0*
Alcohol from ««ss«v«*
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Abstracted i»:0,
A*
^
961«
j|f
1911*
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process®# for production of industrial alcohol,
lag* Cneru. §S* 712.
Ohlason, 1.
Ind.
1933.
I « l*#xist#,iio# d# deux t ©moats amjriolytiques
dans la dlsstiis# da » it*
Compt. **o&, soo* biol. 87.
1183-#4. '1988*
Qphof, A* im
Swelling of omsssm starch.
33, 91^3*
Oha®, Wookblsd*
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1. Soil* Agr* Hokkaido lap. Univ. 19, pt. #*.15$-243*
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Original m % seen*
838S*
1828*
Abstracted in. 0. A. 22.
Dahls*, K*# and Church, if* Industrial sold mmmymm*
lag. 0h«a. |t, 67-71*
Gwen, W* 1*
lad*
1989*
Production of industrial alcohol froa grain b y '
Amyl® process*
Ind* lag* Ghost. If, 89*69* _ 1933*
Petit* fV Brewing, wsterlslii*
129-32.
IfIf*
12, 1680*
Pring®h#ia* 1*
Brasserie at titlharia* f,
Original not **«&•
Abstracted in. 6. A*
If18*
tts afeMdatry of th« sonosaocttarites and. of
th# f©lfsa##terid«s.
McGraw-Hill Book Co.*. I«? York. 1932*
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1oxfcftr !, L* fiat Kfittlo* fU. ?,
Industrial alcohol from cassava.
Philippine Agr. 10, ?S-84.
m m r ma ,. 8*
US,
Alcohol yield.# f r * aelA. saccharified oavoals*
% m m state Soli* 1. Sal* Q ,
£L5~£0*
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1939*
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Taismiae, 1.
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IbxikLo substance and process of making th®
Attests#, ?«
siat*
V* S* Patent 1,4-60,756.
Tubangui, M* A* and others,.
It»S*
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^
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1939*
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e©»i»ris©n of several
lad. lag. ©*.«&, 32, §44-47*
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Separation » M characterization of starch
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Reproduced with permission of the copyright owner. Further reproduction prohibited without permission.
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