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Патент USA US3052557

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United States Patent O-??ce
3,052,547
Patented Sept. 4, 1962
2
1
approximate molar ratio of 5:3:2, and a small quantity
of phosphate. This analysis of frozen beer precipitate is
3,052,547
herewith reported for the ?rst time.
PROCESS OF PREVENTING PRECIPITATION IN
MALT BEVERAGES AND PRODUCT
_
The analytical tests carried out showed that no protein
or lipoidal material was present. The calcium oxalate
was determined both by chelometric titration with diso
Gerhard J. Haas, 170 Pleasant St., Rumford, East Provi
dence, R.I., and Alan I. Fleischman, 89-40 151st
dium ethylenediarnine tetraacetate using acyanide-inhib
Ave., Queens, N.Y.
N0 Drawing. Filed Apr. 7, 1961, Ser. No. 101,343
10 Claims. (Cl. 99-48)
ited eriochrome-T-black indicating at pH 10, for calcium
according to the method used by Owades, I . L., Rubin,
This invention relates to the manufacture of malt bev 10 G., and Brenner, M. W., Amer. Soc. Brew. Chem. Proc.,
193 (1956), and by determining oxalate ion, ?rst con
erages, and more particularly, to a method of treating
verting it to calcium oxide and then acidimetrically ti
malt beverages for avoiding formation of unsightly pre
trating the oxide to a methyl red endpoint according t(
cipitates in said beverages upon freezing and thawing
thereof.
Malt beverages such as beer and ale are not inten
the method used by Burger, M., and Becker, K., Amer
15 Soc. Brew. Chem. Proc., 102 (1949). Although the cal
cium oxalate content in several samples varied, it wa:
found, with anion-free beer, that the presence of calciurr
tionally frozen and thawed. Their freezing point is below
that of water, and usually at approximately 28° F. Nei
ther the manufacturing steps involved in brewing and
packaging malt beverages, nor their transportation and
oxalate was neither a necessary nor a sufficient conditioi
for the formation of frozen beer precipitate. Thus tht
storage require them to be cooled below their freezing 20 calcium oxalate is a coprecipitated impurity in frozel
beer precipitate. The approximate carbohydrate concen
temperature. Still, freezing of a malt beverage occurs
tration was determined by the anthrone procedure accord
not infrequently either under atmospheric conditions of
ing to the method used by Haas, G.'J., and Fleischman
low temperature, or when the beverage is stored in a
A. I., Wallerstein Lab. Comm., 21, No. 73, 193 (1958)
refrigerated space the temperature of which is controlled
Qualitative elementary analysis of the carbohydratl
25
below 28° F.
fraction of frozen beer precipitate produced from anion
It is well established that malt beverages subjected to
free beer showed the presence of nitrogen by sodium
one or several cycles of freezing and thawing may de~
metal fusion, and the absence of halogen and sulfur. Th‘
velop a cloudiness or even a sediment which makes them .
absence of sulfur further con?rmed the absence of pro
unacceptable for consumption. Beer and other malt
beverages are expected to be entirely transparent and 30 tein since wort and beer proteins would have some pro
portion of sulfur-containing amino acid residues as cys
free of solid matter. A precipitate whether suspended in
tine, cysteine, and methionine. Acid hydrolysis followel
the liquid or deposited at the bottom of a container is
by reaction with acidi?ed sodium molybdate and subse
considered undesirable and unappetizing by the customer
quent reduction by acidi?ed ferrous sulfate showed 1th
and evidence of spoilage. Substantial quantities of beer
are returned to breweries by complaining purchasers be 35 presence of phosphate.
By quantitative elementary analysis the presence of ap
cause of the presence of solid material therein after
proximately 1% of phosphate ion and 2.66% of amin
freezing and thawing of the packaged beer has occurred.
nitrogen was found.
Such beer cannot economically be restored to market
Frozen beer precipitate failed to dissolved in cold d.
able condition and is usually destroyed. The loss of the
lute
hydrochloric acid or sulfuric acid and in cold sc
40
goods, and even more important, the loss of customers’
dium hydroxide solution but did dissolve in concer
good will is of substantial importance to the brewing in
trated sulfuric acid. The material gave a positive Molisc
dustry.
test and a positive anthrone test for carbohydrates b1
The precipitate formed upon freezing and thawing of a
failed to give Bial’s orcinol test for pentoses or the phlorc
malt beverage consists of insoluble ?ake-like particles the
nature of which had not been exactly determined here as U! glucinol test for pentoses or galactose. The materiz
failed to reduce copper before hydrolysis but did reduc
tofore.
after hydrolysis indicating it to be a non-reducing pol!
It is one object of the present invention to provide a
meric substance. After hydrolysis, Seliwanoff’s test it
simple and effective method of preventing formation of
ketoses was negative. Reaction of partially hydrolyze
such precipitation and sedimentation on freezing and
thawing.
Another object of the present invention is to provide a
stable malt beverage which is non-precipitating on freez
ing and thawing.
Other objects of the present invention and advanta
geous features thereof will become apparent as the de
scription proceeds.
50
material with phenylhydrazine and sodium acetate yielde
an osazone of indeterminate nature.
A 2.5 hour hydrolysis with 4 N hydrochloric acid in
nitrogen atmosphere in which the ef?uent gases wer
trapped in barium hydroxide solution showed that carbo
dioxide was given off by the formation of barium ca
bonate. A furfural derivative was found in the reactio
?ask by the phloroglucinol reaction. This is indicatii
The present invention is based on the ?nding that the
of the presence of a uronic acid. The presence of uron
insoluble ?akes are essentially of a carbohydrate nature,
acid was confirmed by reacting some partially hydrolyze
and may contain minor amounts of co-precipitated ox
alates if such oxalates are present in the packaged beer. 00 material with sulfuric acid-acidi?ed alcoholic carbazoj
solution to give a characteristic purple color.
Typically, the ?akes consist of about 25% of calcium
The uronic acid was tentatively identi?ed as D-glucu
oxalate and of about 75% of a carbohydrate polymer or
condensation product which, upon hydrolysis, yields glu
cose, N-acetyl glucosamine, and glucuronic acid in an
onic acid by chromatographing partially hydrolyzed m.
terial on Whatman No. 1 ?lter paper, descending, in
3,052,547
3
ahenol-water system and spraying with p-anisidine phos
age. The optimum temperature and duration of treatment
v)hat‘e. Glucose was also found on the chromatogram.
is dependent upon the amount of enzyme used.
The ?-D-glucosaminidase used for carrying out the
method of the present invention may be of any origin, and
D-gIucuronic acid was con?rmed by reacting the partially
rydrolyzed material with mannose and thioglycolic acid
11 the presence of sulfuric acid to form a red solution,
he optical density of which was greater at 410 mp than
it 480 me. Other uronic acids, hexoses, and pentoses have
may be employed either in a puri?ed form or as a crude
extract as far as the impurities of the extract are com
patible with the quality of the treated malt beverage,
?-D-glucosaminidase was found effective for the pur
greater optical densities, in this reaction, at 480 mp. than
it 410 ms. The absence of galacturonic acid, which in
poses of the present invention in the presence of oxalic
nest chromatographic systems has a rate of ?ow (RF) 10 acid decarboxylase previously proposed as a means for
:lose to glucuronic acid, was shown by a negative cysteine
destroying oxalic acid, and may be jointly employed
ulfuric acid test.
therewith for simultaneously achieving the bene?cial re
Glucuronic acid was quantitatively determined by de
sults of oxalate removal.
.
:arboxylating it with 12% hydrochloric acid and deter
While B-D-glucosaminidase in a puri?ed form is effec
nining. the carbon dioxide evolved, titrimetrically. The 15 tive, it is rather uneconomical under present conditions.
arbohydrate fraction of the frozen beer precipitate con
The method of the present invention is preferably per
ists approximately to 20% of polymerized glucuronic
formed with a mixture of carbohydrases effective to by
cid.
drolyze polymers containing glucosamine and produced
Chromatography of a partially hydrolyzed sample indi
by extracting certain edible molluscs, especially the snails
ated that glucose was present. This was tentatively con 20 of the Gastropod class of molluscs, such as Helix pomatia
,rmed by reacting the partially hydrolyzed material with
(the French snail) and sea snails, such as Littorina lit
erric chloride and phloroglucinol and determining its ‘ab
torea (the large periwinkle), Patella vulgata (the limpet),
orption spectrum. The presence of glucose was con
Otala lactea, Helix (helicogena) aperta, and others.
rmed also by the unsulfonated resorcinol reaction ac
ording to the method described by Devor, A. W., Unger,
3. and Gill, ‘I. Arch of Biochem. and Biophys., 72, 20
1958). Employing the presulfonated resorcinol reaction,
Preparations containing ,B-D-glucosaminidase may also
25
be prepared from other sources, for instance, from fungi,
such as Aspergillus oryzae and others.
?-D-glucosaminidase is effective in destroying the
1e frozen ‘beer precipitate was found to consist approxi
precipitate-forming carbohydrate polymer at all temper
iately to 50% of polymerized glucose.
atures between the freezing temperature of the beer and a
‘Chromatography of the partially hydrolyzed material 30 maximum temperature somewhat dependent on the origin
nd spraying with ninhydrin indicated the presence of
of the enzyme.
lucosarnine. This was con?rmed by the Elson-Morgan
:action in which glucosamine is reacted with acetyl ace
me to give, under alkaline conditions, 3-acetyl-2-methyl
-tetrahydroxy butyl pyrrole. This in turn reacted with
-dimethylamino benzaldehyde to give a product absorb
rg at 540 mu. Although the reaction is quantitative for
lucosamine, partial destruction occurs during hydrolysis
f the polymer. Therefore, this test was used only quali
rtively. Glucosamine was quantitatively determined by
re method of Gardell, S., in “Methods of Biochemical
The enzyme is active at a pH normally encountered
in brewing operations, i.e. at a pH between about 4.0
and about 5.5.
The enzyme activity is substantially a linear function
of enzyme concentration within the concentration range
which is practical for the purpose of the present inven
tion. Even small amounts of ?-D-glucosaminidase have
a noticeable effect in reducing the amount of sediment
40 formed in beer upon freezing and thawing. There is, of
rnalysis,” vol. 6, p. 302 (1958), Interscience, N.Y., in
him hexosamines are quantitatively deaminated under
lkaline conditions in a phosphate-borate mixture and the
mino nitrogen determined by nesslerization. This method us Or
'ill not determine amino acid, protein, or amide nitro
en. The precipitate had approximately 27% of poly
rerized glucosamine.
Since glucosamine is often found acetylated, a quantita
ve acetyl determination was run. It was found that the
alymer carbohydrate contained 5% of acetyl groups.
From these results it is concluded that the carbohy
rate polymer consists of glucose, n-acetylglucosamine,
ad D-glucuronic acid in the ratio of 5:3:2.
~
In a beer treated for removal of oxalate ions the per
:ntage of calcium oxalate in the insoluble ?akes is lower
course, an upper useful limit determined by the maximum
quantity of the polymer carbohydrate that may be en
countered in beer under normal conditions of commercial
practise. This limit was found to be at a concentration
of about 120 ?-D-glucosaminidase units as determined ac
cording to A. Linker, K. Meyer and B. Weissmann, “J.
Biol. Chem.,” vol. 213 No. 1 (1955) per 1000 cc. Of
beer or process liquid when using an enzyme prepara
tion as obtained according to Example 1 given herein
after. Such a concentration of enzyme is more than
adequate under usual commercial conditions, and no use
ful purpose is served by exceeding it.
The following examples may serve to illustrate the
present invention without, however, limiting the same to
the speci?c embodiments chosen for the purpose of the
disclosure.
ran 25%, and may actually be too small to permit re
EXAMPLE 1
Fifty edible sea snails of the species commonly known
The origin of the polymeric substance has not been
as brown-shelled edible snails (Otala Iactea) were dis
:termined with certainty. It is found in unfermented 60 tended by submerging them in Water. They were then
ort consisting solely of malt and water, and treatment
mechanically deshelled and their ?eshy portions were
E such wort by the method which will presently be
ground with ice water for 2% minutes in a laboratory
sclosed is sufficient to prevent formation of an objec
mill of the “Omnimixer” type whereby the snails were
Jnable precipitate in a packaged beer produced from
macerated and extracted. The ground mixture was cen
trifuged. The supernatant liquid extract was decanted
e wort and subjected to many cycles of alternate freez
g and thawing.
and amounted to 190 cc. The solids were discarded.
Solid ammonium sulfate was dissolved in the extract
In principle the method according to the present inven
able quantitative ‘estimation.
Jn consists in the addition of an enzyme preparation
mtaining ?-D-glucosaminidase to the wort, to the ?nished
xr prior to packaging, or at any intermediate stage of
e brewing process. Such addition has proved to be
ghly effective in degrading the carbohydrate polymer
hich is responsible for formation of a solid precipitate
ion freezing and thawing of the packaged malt bever
until the solution had a concentration of the added salt
corresponding to 40% saturation while the temperature
70 was held at 34° F.
A precipitate formed and was re
moved by centrifuging.
Ammonium hydroxyde was
added to the liquid to adjust the pH to 6.5 and additional
ammonium sulfate was dissolved therein to raise its
concentration to 50% saturation. The precipitate formed
was recovered from the liquid by centrifuging and was
3,052,547 -
5
6
After the ninth cycles ___________________ __g__ 0.049
dissolved in an 0.1 molar sodium acetate buffer solution
adjusted to a pH of 4.5. The solution ‘was dialyzed
After the tenth cycle ____ __' ______________ __g__ 0.056
against an 0.1 molar phosphate buffer solution having
a pH of 7.0 for 20 hours.
EXAMPLE 3
"
5 mg. of the precipitate recovered by ?ltration in Ex
ample 2 were dispersed in 2 cc. of snail enzyme solution
obtained according to Example 1, the pH of which was
The non-dialyzable fraction contained 1.3 mg. per cc.
of protein and had'a ?-D-glucosarninidase activity of
15.4 units per cc.’ This solution will be referred to here
inafter ‘as snail enzyme solution. This solution may be
4.5, and in which the precipitate was insoluble. The re
sulting mixture was inculcated at 99° F. ‘for 18 hours.
converted into the dry state by lyophilizing.
v10 The solid material dissolved. The presence of d-glucose,
An analysis of lyophilized snail enzyme gave the fol
' of N-acetyl glucosamine, and of d-glucuronic acid in the
lowing results.
solution was con?rmed by chemical tests and by chro
A. Chemical Analysis
matographic analysis, as described above.
Component:
I
Percent
EXAMPLE 4
-
Moisture
____________________________ __
'Lipids
_
Total protein
_____
' 12 oz. portions of beer from the same batch from
10.05
which the cans of Example 2 were taken were mixed
___-
Hexoses, polymerized __________________ __
‘Ash
'
15
6.45
___- 20.00
'
2.15
61.60
Uronic acids ______ _'_, _______________ __'__
1.04
Hexosamines ________ _ '_ _______________ _ _
0.5 1
B. Enzymatic An‘alysis
Enzyme
Units/g.
?-Glucuronidsse _______________ ._
TABLE
Reference
22, 800
Amylase ______ ._
with snail enzyme solution and permitted to stand at 8°
C. for 5 days. The mixtures were packed in metal cans
20 identical with' those used in Example 2, and were sub
jected to ten cycles of the afore-described freezing and
thawing treatment. The cans were then opened and ex
amined. The results are tabulated below.
25
Enzyme concentration (units/liter):
0
2
Cpllnlnsn
1,550
3
B-Glucosaminidase ____________ _.'____________ _.
6, 000
4
30
References describing the units:
Result
_______________________ __ Heavy ?akes (+).
6 _______________________ __ Clear (neg).
1
Fishman Unit. “Advances in Enzymology,” vol. 16, pp.
955).
Bernfeld, Edited by S. P. Colowick and N. 0. Kaplan (1955),
. Methods in Enzymology, vol. '1, p. 149, Article by P.
11 __________ __' __________ ..-
D0.
36 ______________________ ..
60 ______________________ __
95 ______________________ __
Do.
Do.
Do.
119-.. _____ _‘_ ____________ .._
1
Do.
That the husk is not the prime source of the polymer
Academic Press N.'Y.
3. ‘Myers, $15.11., & Northcote, D.H.; Biochem. J., vol. 71,
p. 749 (1959). 1 unitzthat quantity of enzyme giving 1 35 causing frozen beer precipitate, was proved by the follow
microgrum glucose at 25° C. _in 12 hours from insoluble phos
ing test.
>
phoric _acid swollen eotten lmtners at pH 5.6.
_
Beer was brewed from dehusked malt by standard
4. Linker, A., Meyer, K., ‘and Weissmann -B., J. B101. Chem,
American practice on a pilot plant level. The beers were
vol. 21-3, No. 1 (1955). 1 unit is defined as that quantity
of enzyme which liberates 1 microgram of phenol in 1 hour
canned, pasteurized, and frozen and thawed. When ex
at 37° C. at pH 4.6 from N-acetyl-B-phenyl-D-giucosaminide
which was synthesized according to the method of Helferrch, 40 amined, ?akes of frozen beer precipitate were found.
13., and 11011, A., J. Physiol. Chem, vol. 221, p. 252 (1933).
By the above mentioned treatment with the enzyme so
EXAMPLE 2
A commercially produced Lager beer packed in con
, lution formation of the precipitate was completely pre
vented.
.
EXAMPLE 5
ventional .12 oz. cylindrical metal cans was subjected to
alternate storage in air at —l8° C. and at +24° C. for 24 45
12 oz. portions of beer from a similar batch from
hours each. The beer as received had the following prop
which the cans in Example 2 were taken were mixed
erties characteristic of good commercial practice:
with snail enzyme solution and oxalic ,decarboxylase.
Average
Color, V2" cell, Lov. Ser. 52 _______________ __
Apparent extract, percent __________________ ___
Real extract, percent _______________________ __
They were permitted to stand at 8° C. for 5 days, then
The cans were
2.6 50 commercially canned, and pasteurized.
subjected to ‘the aforementioned freezing and thawing
2.99
4.73
treatment and then examined. The results are tabulated
below.
Alcohol by weight, percent _________________ __ 3.78
Original extract, percent_________________ __>__ 12.02
Degree of attenuation, percent ______________ __ 60.8 55
Reducing sugars, percent __________________ ___‘Protein, percent
___
1.21
0.31
Acidity (as lactic), percent___>_‘_ _____ ___; _____ __
0.14
pH
..
__
_
_
TABLE
Enzyme
Oxalic de-
tion
eoncentra- formation
(units/liter)
tion
4.25
The storage periods were 24 hours to cause complete
freezing of the can contents during —18° C. storage, and
also 24 hours to'cause complete thawing at +24° C.
Sample cans weredrawn before the ?rst cycle and after 65
the ?rst, third, ?fth, and tenth cycle of alternate freezing
and thawing. The contents of the sample cans were re
0
0
6
11
36
60
95
321
321
321
321
321
neg.
neg.
neg.
neg.
neg.
100
100
100
100
100
119
321
vneg.
100
600
321
neg.
100
standard Americancommercial methods, graded amounts
of snail enzyme (solution were added. The resultant beer
After the ?fth cycle __________ "I ________ __g'__"0.026
_
c
' 0.028
Afer the seventh cycle ______ __, _________ _.'.__g__ 0.034
‘
Wort was prepared by the usual commercial procedure.
Just prior to the addition of yeast and fermentation by
After the third cycle ____ __‘.... ___________ __g__ 0.014
'
0
' EXAMPLE 6
'
Before the ?rst cycle ______ _..__,__..; ______ __ _Brilliant
After the'?rst cycle ____ __.._.. ____ __'___..___gv_.. 0.010
~_
+
‘ Oxalic acid was determined mauometrlcaliy.
moved and ?ltered. The weight of material retained by
After the sixth cycle
oxalate
destroyed
(units/liter)
Carbon dioxide, volumes ___________________ __ 2.67 60
Air content, ml./ 12 oz ______________________ __ 1.1
the ?lter was determined to be as follows:
'
concentra- carboxylase Precipitate Percent of
was treated as described in Example 4 and subjected to
75
the same tests.;
'
'
.
3,052,547 '
TABLE
-
' days, canned, commercially pasteurized, and frozen and
vthawed. The results are shown below.
Frozen beer
Units of enzyme added per liter:
0
precipitate
+
0.8
2
TABLE
Neg.
Neg.
4
‘
8.
24
_
,
Units of Snail Enzyme Added per Liter
Frozen Beer
Residual
Precipitate
Formed
Neg.
Oxalic Acid
(Parts per
Million)
Neg.
Neg.
'
40
78
0
8- 7..-"
Neg.
Neg.
+
Haze :l:
.
17. 4
0
_
0
0
0
A very slight‘haze was seen in the treated material
when examined under strong lights, while heavy ?akes
were seen in'the control.
EXAMPLE 1 1
Chemical analysis indicated
that the haze consisted primarily of calcium oxalate and 15
proteinaceous material. Carbohydrate (Molisch test)
Several brands of American beers bought in the New
York area were treated with 26 units of snail enzyme
and uronic acid (carbazole test) were absent. The haze
l3-glucosaminidase per 700 cc. of beer and 244.7 units of
oxalic decarboxylase. The beers were stored for 3 days
immediately dissolved in normal hydrochloric acid (un
like frozen beer‘precipitate) and upon hydrolysis gave
at 8". C., canned, and pasteurized by commercial meth
only amino acid tests (for instance, Hopkins-Cole gly 20 ods. The cans ‘were subjected to alternate ‘freezing and
oxalate test etc.).
The oxalate was therein a major con- I
stituent and was decomposed to carbon dioxide by a treat
ment with oxalic decarboxylase in a Warburg respirom
eter.
t
'
thawing-and examined for the formation of frozen beer '
precipitate. The ‘beer formed the precipitate Without
treatment but did not form the precipitate when treated.
-
EXAMPLE 12
25
EXAMPLE 7
To 700cc. of an imported all malt beer there were
An enzyme preparation obtained from a crude extract
of Aspergillus‘ oryzae as sold under the trademark
added 11 units of snail enzyme (}3~glucosaminidase). The
sample was stored at 8° C. for 3 days, canned, pasteur
per gram was added to beer in an amount of 100 mg. of 30 ized, and frozen and thawed.
The untreated control showed heavy ?akes. The
“Mylase-P” per liter. The mixture was allowed to stand
treated sample was slightly hazy but devoid of ?akes of
at 35° C. for 24 hours. vThe samples were frozen and
frozen beer precipitate.
thawed in polyethylene containers and examined for the
The process according to the present invention can
formation of frozen beer precipitate. The control sample
showed precipitate while the treated sample was devoid 35 also be employed in the known process of producing
concentrated malt beverages such as beer by freezing.
of it.
'
“Mylase-P” containing 24,200 units of glucosaminidase
Enzymatic analysis of “Mylase-P” showed high con
centrations of the enzyme ?-D-glucosaminidase and was
devoid of p-glucuronidase or cellulase.
EXAMPLE 8
A mixture of the enzyme preparation from snail ex
tract containing 5 units of B-glucosaminidase and 321 units
Addition of p-D-glucosaminidase before such freeze-con
centration considerably reduces the formation of precipi
tates in the concentrated beverages.
Of course, many changes and variations in the prep?
40 aration of the ?-glucosaminidase employed, the amounts
of enzyme added, the reaction conditions, temperature,
and duration, and the like may be made by those skilled
in the, art in accordance with the principles set forth here
of oxalic decarboxylase was added to 1000 cc. of wort at
in and in the claims annexed hereto.
the start of a normal fermentation. ‘As a result of this 45
We claim:
_ addition both the formation of frozen beer precipitate and
1. In a process of preventing formation of precipitates
the formation of calcium oxalate haze in the beer were
in fermented malt beverages, the step which comprises
materially prevented.
adding at least at one stage of the brewing process, prior
to the consumer packaging of the beverage, an enzyme
EXAMPLE 9
50 preparation containing B-D-glucosaminidase to the brew~
ing liquids in an amount corresponding to at least 0.8
unit of ,B-D-glucosaminidase per liter of beverage, so as
to prevent formation of precipitates on freezing and thaw
Several batches of wort were prepared by the usual
commercial procedure. Just prior to the addition of yeast
and fermentation by standard American commercial
methods, graded amounts of snail enzyme solution'were
added. ‘The resultant beer was treated as described in
Example 4 and subjected to the same tests.
_
I
-
0.8
2.4
4.0
8.1
24
8O
160
.
malt beverage.
‘
3. The process according to claim 1, wherein the brew
ing liquid to be treated is the 'wort.
Precipitate formation
--
-
2. The process according to claim-1, wherein the brew- _
ing liquid to be treated is a substantially end-fermented
TABLE
Umts enzyme added: 0
ing of the beverage. '
--
+
60
.
4. The process according to claim 1, wherein the brew
ing liquid to be treated is‘ ‘the fermented malt beverage.
Neg.
Neg.
5. The 1 process according to claim 1, wherein the
enzyme preparation added to the brewing liquid is an
Neg.
enzyme preparation obtained from sea snails.
Neg.
Neg. 65 6. The process according to claim 1, wherein an oxalic
acid-destroying enzyme is additionally added to the brew
Neg.
ing liquid.
~
Neg.
7. The process according‘ to claim 1, wherein the‘
enzyme preparation added to the brewing liquid is an
70 enzyme preparation obtained from fungi.
V
EXAMPLE 10
8. The process according to claim 1, wherein the
To ale, prepared by standard American commercial
enzyme preparation added to the brewing liquid is an
practice, graded amounts of snail enzyme (?-glucosamin
enzyme preparation obtained from Aspergillus oryzae.
idase) plus 224 units of oxalic decarboxylase per liter
9. In a process of preventing formation of precipi
were added. The samples were stored at 8° C. for 3 75 tates in fermented malt beverages, the step which com—
No ~?akes were seen except in the untreated control.
3,052,547
prises adding at least at one stage of the brewing proc
ess, prior to the consumer packaging of the beverage,
?-D-glueosaminidase to the brewing liquids in an amount
corresponding to at least 0.8 unit of B-D-glucosaminidase
per liter of beverage, so as to prevent formation of precipi- 5
tates on freezing and thawing of the beverage.
10. In a process of preventing formation of precipi
tates in fermented malt beverages, the step which com
prises adding at least at one stage of the brewing process,
prior to the consumer packaging of the beverage, 5-D 10
10
glucosaminidase and an oxalic acid-destroying enzyme vto
the brewing liquids in an amount corresponding to at
least 0.8 unit of ,B-D-glucosaminidase per‘ liter- of bever
age, so as to prevent formation of precipitates on freezing
and thawing of the ‘beverage.
References Cited in the ?le of this patent
UNITED STATES PATENTS '
2,878,125
Brenner ___________ __'__ Mar. 17, 1959
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