Note : Les descriptions sont présentées dans la langue officielle dans laquelle elles ont été soumises.
-- 2 --
HOE 8Z/F 275
It has already been proposed that functionaL
hydrolysates of microbial protein ;solates be prepared
by initially subject;ng a m;crob;al protein to an extrac-
t;on treatment to reduce the content of nucleic acids and
l;p;ds in it, and then subjecting it to enzymatic hydro-
lysis by one or more endoproteases (Canadl~n Patent
Application No. 414,860).The protein hydrolysates have
advantageous properties in the technoLogy of foodstuffs
and bring about not only a nutritional improvelnent in
foods, but also facilitate the ;ndustrial processing of
them, especially if they have a porous structure but, at
the same time, contain ~ater-soluble and lipid~soluble
constituents, as is the case ~ith desserts, baked pro-
ducts and pasta, and imitation cheeses~ One fraction of
these protein hydrolysates shows a particularly high
emulsifiability as is required for sausage products~
cheese products, confect;onery and creams~ but aLso for
mayonnaise and salad sauces or other fat preparations.
It has no~ been found that spec;fic protein
hydrolysates of this type are able to serve not only as
food additives contained in foodstuffs to only a minor
extent, but rather as protein components ;n hi~h-protein
foodstuffs~ especially ;n cheese-like products.
The invenLion relates to 3 process for the pre-
paration of a functionaL hydroLysate of microbial protein
~ 3
isolates by extraction of a micrchial protein to decrease
its content of nucleic acids and lipids~ follo~led by enzym-
atic hydrolysis by endoproteases, which process comprises
carrying out the hydrolysis with physiologic proteases under
mild cond;tionsf and relates to the functiol7al protein hydro-
lysate-s obtainable thus and to preparations which contain
these functional prote-in hydrolysates~ Preferred embodi-
ments of the invention are illustrated in more detail below.
All h-igh-protein microb;al cell aggregates are
suitable as the startin~ material. The bacteriaL cell
aggregates whicli are obtained according to Gerrnan Patent
2,633,451 by culturing bacteria of the strain Methylomonas
clara ATCC ~1226 in a nutr;ent medium containin0 methanol,
out of which acigre-~ates the lipicls have essentially been
rcmoved by the process in German Auslegeschrift
2,h33~6~6, by extrac-tiorl with a mixture, which is larciely
or completely anhydrous, of ammonia and a polar solvent
from the series of lower alkanols; lo~ler glycols or
methyl or ethyl ethers of a lo~er glycol, in particular
nethanol, and the nucle;c ac;ds llave essentially been
removecl in a subsequellt step by extraction uith water~r
are particularly advant-ageous.
Trypsin~ chymotrypsin and mixtures thereof are
pre-ferred as the physiologic endoproteasesr Trypsin is
particularly preferred. These endoproteases lead to a
product havil~g a satisfac10ry flavor and valuable phys;o--
logic and industrial propert1es. It is possible to use
"aggressivc" en~yl,lesr hut this requ-ires the main-
ten~nce and r,icnitcring oF particularlv adjuste~ cond;~
r~ ~ e
~ 4 ~
tions, this not bcing generally advantageous by reason
of the increased elaboration. Enzymes such as pepsin
hich cleave in a very acid range lead, when neutral;zed
as is subsequently necessary, to production of an inc-
reased amount of salts which can be disadvantageousO
- Obv10llsly, the hydrolysis conditions are governed
by the enzyme employed. In the case of the preferred
proteases~ hydrolysis takes place in the ptl range 7 to 8,
in particular at pH 7.5, in a ternperature range from ~0
10 to 50, in particular 45, C, over the course of 5 to 30,
in particular about 20, minutes~ In general, 500 to
2,000 parts by weight, in particular about 1~0L10 parts by
we;~ht, of substrate are employed per ont part by weight
of protease.
When the hydrolysis has reached the required
extent, the reaction is stopped, advan.ageously by
inactivating the enzynle. This ;s advantageously carr;ed
out by heating the batch to 70 to 90, ;n part;cular 80,
C wh;ch takes a few M;r~utes.
The funct;onal protein thus obtained can be
dried~ for example in a spray dryer, and it can then be
stored. Whtn this dried material is used~ it is first
resolvatized, this advantageously taking place under
slit~ht vacuu~ in order to prevent inclusions of air~
However, it is advarltc1geous to process the pro-
tein hydrolysate d;rectly to give the applopriate pre--
paration~ Thus~ the hydrolysis can be carried out, for
exampief in a processed cileese kettle after which tht
product is ir~mediately further processed with the cus-
i ~cooker)
~Z~77~ .
~ 5
tomary adclitives~ such as emulsifyin~ salts~
A number o-f processes~ in which the proportion
of cheese used in processe~ cheese and simllar products
is wholly or partly replaced by proteins wh;ch are more
5 readily manipulated industrially and are nlore -favorably
pr;ced-, have already been disclosed. ~xamples of this
are unriperled milk protein (Britisl1 Patent 1,601~672,
German Offenlegungsschrift 2~834,093), caseinate (German
Offenlegungsschrift 2,4-11,913, U.S. Patent ~,2~2,050) or
products from whey (U~S. Patent 4,166,1(~2), some o~ them
being used a~ter mixing with vegetable products, such as
wheat gluten (U.S~ Patent 3~806,606),. soybean protein
tlJ.S. Patent 4,080,477) or peanut prote-in (I:ood Techno-
Lo~y, July 1979, 88 - 93).
The use of a microbial prote;n, a sodiurn, calciuln
or magnesium salt, an acetylation product or a phosphate
complex thereof or a l~eat-denatured microbial protein as
the protein comporlent in foods, such as baked products,
meat products, drinks, -fermented dairy substit;ute pro-
ducts or desserts has been disclosed in German Offen-
le~un~sschriFt 2,651,464.
It is important, ~or health and flavor reasons~
especially with protein isolates from bacteria, to
decrease the cor~ent of nucleic acids and lipids. How-
ever~ ~from the industrial v1ewpo-,nt, isolates of this
type cannot directly be employed for ,oodstuffs~
If it is at.tempted, for example; to employ a
protein isolate whicl1 ~as been prepclred by the Metl,od of
German ~uslegeschrift 2~633,66S for a processed cheese
6 --
like productt dif,iculties arise e~en when high propor
tions of elnulsifying salts are usedr a homo~eneous and
smooth emulsified composition is not obtained, and this
is evident not only from the crurnbly and mealy cons;st-
S ency, but it aLso leaves a gritty sensation like sar?cl inthe mouth.
t-lowevel, surprisingly, the fun~ctional bioprot2ins
according to the invention ar excellently suitable as
protein components for cheese substitute products. On
1U processing with customary emulsify1ng salts as ~re des-
cribed in U.S. Patents 4,080,477 and 4,166,14Z, German
Offenlegungsschriften 2,411,913 and 2,834,093 and British
Patent 1,601,672~ ~ood protein breakdown, c~ood emulsifi-
cation and a good texture are achieved, high-polymeric
polyphosphates being preferred. P~eme1tabi1i-ty is
achieved by mixtures of citrate and polyphosphate.
On manufacturin~ processed cheese-like products,
it is possible to add othr protein carriers, such as
cheese or the replacement products which have been men
tioned. Butter fat and vegetable fats are suitable as
the fat component. It is possible to add, in a customary
manner, water an(3 auxiliaries, such as emlllsi~iers~
colorants and flavorings.
The products can be procssed in cuscolTIary
Z5 machines (for exarnple pan,
cutter or 1aydolln cooker) and filled out ;n a custor(lclry
manner and packed, for example for spread;ng, ;n alu-
m;num foil~ tubes, tubular film, jars or plastic tubs
or, for slicing~ in blocks~ It is also poss;ble to
3'7~
- 7 -
endles s
process them to produce slices in known~tuber bel~ or
roller processes~
In addition, it is possibler using the ~-unc.^;Gnal
bioproteins accordin~ to the invent;on, to manufacture
other preparations, such as, for example~ spreads, coffee
wllitcne*s, sauces or the like.
In the examples which follow~ ~percentage data
relate to weight unless otherwise spec;fied. The addi-
tives are characterized in compliance with the regula-
tions ~overning the l;cens;ng of additives (ZZulV~ BGBl
T.I r 1~81~ 1033).
*German
E _ ple 1
Preparation of a functional bioprotein accordin~ to the
i n v e n t :l o n .
The starting material used is a bacterial cell
aggrc~ate according to German Patent 2,633,~S1, Cxample
2, the content in whicl1 o~ nucleic acids and lipids beirlg
decrcased by the process in German Auslegeschrift
2~633,6O6, Example 1.
10 kg of this protein isolate are suspended in
65 liters of ~Jater and the pH of the suspension is
adjusted to 7~5 by the addition of 4 N sodium hydroxide
solution.
The suspension is preincubatecl at 45C with
25 stirring, tl1en 20 ~ or tryps-in PTN 3.0 S (supplied by
~ovo) are cadded and ;ncubat;on is carried out, wi'ch
s-tirring, at 45~` for 20 min. To ;nacc'ivate the enzyme,
the batch is heated to ~0C ~or 10 min arld the suspensiQn
is spray-dried'. 1'he p~l o, the product falls to values
7~
. ~
hetween ~.5 and 7~0 because of the hyclrGly~cic activity
of the enzyme~
e_2
Production of analogs of slices of processed
cheese containin0 50% fat in dry matter, and 56~ dry
matter. The following recipe is used:
15.000 kg of rennet casein
5.000 kg of spray~dried hydrolyzed bioprote;n lsolate
accord;ng to Example 1
30.000 kg of butter
3.000 kg of sk;mmed mil~ powder
0.600 kg of sod;um chloride
1.350 kg of comnlercial emlllsify,ng salt m;xture baseci on
sodium polyphosphates ~E 450C), socdiurn citrates
~E 331), potassium monophosphates tE 340),
sodium diphosphates (E 450 a), sodium triphos
phates ~E 450 b) and sodium rnonophosphates
(E 339)
O~OS0 kg of ci~ric acid
2U 0.200 kg of cheese colorant
32.nG0 kg o~ water~ includin~ condeilsate~
; h~c~le~st~
wh;ch is provided with a stirrer at 120 rprn~ is raised
to 80C l~ithin 7 min.
The product thus produced ;s characterized by a
rat;o of bioprote;n isolate to rennet casein o-r 1 : 3.
The sl;ces show good elastic properties and good toasting
characteristics.
7~
Fxam~le 3
Example 2 is repeated with the modificat;on that
25.000 kg of hydrogenated vegetable fat is ernplo~ed in
place of the 30O000 kg o-f butterr and the armount of ~ater
is increased to 37.000 kg. A product havins the same
prope~t;es is obtained.
Example 4
Production of a product as in Example 2, but
using the f Q l lowi ng recipe:
9.750 kg of rennet casein
9.750 k9 of spray-dried hydrolyzed bioproteirl isolate
accordiny to Exarnple 1
30.000 kg cf butter
- 3.000 kg of skinlrned milk powcler
0.600 kg of sodium chloride
1~350 kg of emulsifyin~ salt (as in Exarrlple 2)
0.050 kg of citric acid
0.200 kg of cheese colorant
32.000 kg of water including condensate.
Z0 The heating is carr;ed out as described in Example 2a
The product thus produced is character-ized by a
ratio of bioprotein isolate to rennet case;n of 1 o 1.
The slices stlow good elastic properties ancl
moderate to ~tood toast;ng characteristics.
25 Example S
ExaMple 4 ;s repeated wi-th the modification that
25~000 kg o-f hydrogella;ed ve3etable fat are employed in
place of the ~n.ooo k~ of butter, the arlount of water
being incre;ised to 37nOOO k~ A product tlaving t}l~ san;e
2~
- 10 -
properties as in Example ~t iS obtained.
Production of analogs of processed cheese for
spreading, containing 40% fat in dry rnatter and 41%
dry matter. The following recipe is used:
3.000 kg of rennet casein
9.000 kg of spray-dried hydrolyzed bioprotein isolate
according to Example 1
1 2 . 0 0 0 k 9 of butter
2.000 kg of skimmed milk powder
0.400 kg of sodium chloride
1.300 kg of commercial ernulsifying salt based on sodium
polyphosphates (E 450c)
2.400 kg of precooked cheese
5 33.500 kg of water including condensater
~cheese
The temperature in the~processing equipment, which is
provided with a stirrer at 120 rpm, is raised to 92C
~ithin 15 min.
The product thus produced is characterized by a
20 ratio o-f bioprotein isolate to rennet casein of 3 : 1n
The cooled melt has ~ood spreading character;s-
t;cs and exllibits an acceptable cheese-like behavior in
the mouth.
e 7
~_ . .
Exanlple 6 is repeated with the modificat;on that
9.6 kg of hydrogenated vegetable fat are employed in
place of the 12.000 kQ of butter, the arnount o-f water
be;ng ;ncreased to 35 900 kg. A product ha~lin~ the sar~e
properties as in Example 6 is obtainedn
7~
Exam~
Production of analogs of processed cheese for
spreading, containing 60% fat in dry matter, and 47% dry
matter. The following recipe is used:
3~000 kg of rennet casein
9.000 kg of spray-dried hydrolyzed b;oprotein isolate
according to Example 1
Z.000 kg of skimrned milk powder
23.000 kg of butter
1.500 kg of ernulsifying salt (as in Example h)
3.600 kg of precooked cheese
37.000 kg of water incLuding condensate.
The heatir,g is carried out as described in Example 6.
The product thus produced is characterized by a
ratio of bioprotein isolate to rennet casein of 3 : 1.
The cooled cheese has good spreading characteris-
tics and exhibits a pleasant cheese-like behavior in the
mouth.
_xample 9
When the process is carried out as in Example 8r
but 22.400 k~J of hydrogenaced vegetable fat are employed
in place of the Z~000 kg oi butter, and the amount of
water is increased to 43.400 kg, then a product having
the same properties as in Example ~,is o,btained.
Example 10
Production oF a high-protein spread with a peanut
flavor containing 25% fat in dry Ina~cter, and -SS% dry
matter.
The follol~ing recipe is used:
~..2~ 7'â~2J
- 12 -
6.000 kg of skimmed milk powder
6.000 kg of spray-dried hydroLyzed bioprotein is~late
according to Example 1
O .400 kg of sugar
12.000 kg of peanut butter
0.430 ky of commercial emulslfyirlg salt mixture based on
sodium polyphosphates ~E 450c), sod;um mono-
phosphates (E 339), sodium diphosphates (E 450a)
and sodium triphosphates (E 450b)
0.05 kg of carboxymethylcellulose (E 466)
49.000 kg of water ;ncluding condensate.
The heating is carried out as described in Example 6.
The cooled melt has a good cons;stency and, com-
pared with peanut butter~ has good spreading characteris-
tics even in the cold state. The product has a pleasantpeanut flavor.
Example 11
Production of a high-protein spread with a peanut
flavor and contain;ng 45X fat in dry matter, and 44% dry
matter. The following recipe ;s used:
3.000 kg of skimmed milk powder
3.000 kg of spray-dried hydrolyzed bioprote;n -isolate
accordillg to Example 1
6.000 kg of butter
0.200 kg of suyar
6.000 kg of peanut butter
0.220 kg of emulsify;rlg salt (as in Exalnple 10)
0~020 kg o-f carhoxymethylcellulose (E 466)
Z0.600 kg of water including conderlsaten
t
- 13 -
Heating is carr;ed out as described in EY~ample 6.
The cooled melt has a good consistency and, com-
pared with peanut butter~ has good spreading characteris-
tics even in the cold state. The procluct has a pleasant
peanut -flavor.
Exampl~e 12
When the process is carr;ed out as in Example 11,
but 4.800 kg of hydrogenated veyetable fat is used in
place of the 6~000 kg of butter, and 21.800 kg ;s used
in place of the 20.600 kg of ~later, then the properties
of the product correspond to those of that in Example 11.
Exarnple 13
Production of a coffee whitener.
The following recipe is used:
800 g of water
70 g of vegetable fat
120 g of corn syrup
60 9 of spray-dried hydrolyzed bioprotein isola-te
accord;ng to Example 1
0 8 g of phosphate stabilizer mixture consistin~ of potas~
sium rnonophosphates (E 34U), sodium polypllosphates
(E 450c) and sodium monophosphates (E 339)~
The pl~osphate stab;l;zer m-ixture ;s d;ssolved in
the water, and then the other constituents are added and
the mixturc is heated, homogenized and spray-dried.
The product clistr;butes well 7n coffee and
exhibits ~Jood wllitellinl3 properties.
