CN1212072C - Activation of hydrolase for proteins in seeds of oil crops and hydrolysis for vegetable protein - Google Patents

Activation of hydrolase for proteins in seeds of oil crops and hydrolysis for vegetable protein Download PDF

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CN1212072C
CN1212072C CNB021573972A CN02157397A CN1212072C CN 1212072 C CN1212072 C CN 1212072C CN B021573972 A CNB021573972 A CN B021573972A CN 02157397 A CN02157397 A CN 02157397A CN 1212072 C CN1212072 C CN 1212072C
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hydrolysis
raw material
activator
soybean
enzyme
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CN1509626A (en
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鲍鲁生
陈雪松
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BEIJING FOOD RESEARCH INSTITUTE
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BEIJING FOOD RESEARCH INSTITUTE
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Priority to PCT/CN2003/001095 priority patent/WO2004056980A1/en
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    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/14—Hydrolases (3)
    • C12N9/48—Hydrolases (3) acting on peptide bonds (3.4)
    • C12N9/50—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25)
    • C12N9/63—Proteinases, e.g. Endopeptidases (3.4.21-3.4.25) derived from plants
    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12Y—ENZYMES
    • C12Y304/00—Hydrolases acting on peptide bonds, i.e. peptidases (3.4)
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
    • A23C11/00—Milk substitutes, e.g. coffee whitener compositions
    • A23C11/02—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins
    • A23C11/10—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins
    • A23C11/103—Milk substitutes, e.g. coffee whitener compositions containing at least one non-milk component as source of fats or proteins containing or not lactose but no other milk components as source of fats, carbohydrates or proteins containing only proteins from pulses, oilseeds or nuts, e.g. nut milk
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/14—Vegetable proteins
    • A23J3/16—Vegetable proteins from soybean
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
    • A23J3/00—Working-up of proteins for foodstuffs
    • A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
    • A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
    • A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
    • A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
    • A—HUMAN NECESSITIES
    • A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
    • A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
    • A23L11/00—Pulses, i.e. fruits of leguminous plants, for production of food; Products from legumes; Preparation or treatment thereof
    • A23L11/60—Drinks from legumes, e.g. lupine drinks
    • A23L11/65—Soy drinks

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Abstract

The present invention relates to vegetable protein and the field of the deep processing of vegetable protein products, which provides a method for activating proteolytic enzymes in oil-bearing crop seeds and a method completely different from the traditional process for hydrolyzing vegetable protein, particularly a method for activating and utilizing proteinase in the oil-bearing crop seeds. The present invention also provides vegetable protein products, particularly instant soybean milk powder and soybean milk powder without fishy odor, produced by the method; the present invention not only reduces the high cost of proteinase preparations used for proteolysis, but also effectively eliminates the fishy odor of soybean of products of soybean milk powder, etc., and the solubility of the soybean milk powder is improved and enhanced.

Description

The hydrolysis of the activation of proteolytic enzyme and vegetable protein in the oil crops seed
Invention field
The present invention relates to the deep processing field of vegetable protein and plant protein preparation, relate in particular to research the contained proteolytic enzyme of oil crops self.The invention still further relates to the method for hydrolysis of the vegetable protein in the described oil crops seed, it has adopted the technology that is different from prior art fully, do not use expensive protease preparation, but finish hydrolytic process by the protease of suitable condition activation oil crops self.The invention still further relates to the product, the especially soy protein products that utilize this method for hydrolysis preparation.
Background of invention
All containing rich in protein in the oil crops seed, is the main source of the essential phytoprotein of the mankind.If use proteolytic enzyme can improve yield in phytoprotein processing, the leaching process, change the various materializations of protein, biological nature, develop new product, enlarge the range of application of phytoprotein, this has been insider's a common recognition.The processing of phytoprotein, leaching process are the processes that protein macromolecule wherein is dissociated into small-molecule substance, for example by being controlled at different hydrolysis stages, can partial hydrolysis become polypeptide, or complete hydrolysis become micromolecular amino acid.In three kinds of hydrolysis methods that utilize acid, alkali and enzyme, enzymolysis is a comparison mode gentle and that control easily, so when requiring hydrolysate to be peptide, usually select the enzymolysis mode, promptly selecting suitable protease preparation that protein molecule is cut off under certain condition becomes little molecule (peptide or amino acid).Oil crops are main sources of vegetable protein, can be extensive use of proteolytic enzyme in its deep processing and the new product development, but the commercially available protein enzyme preparation costs an arm and a leg, and have limited the exploitation of plant protein products.Profound influence will be brought to the vegetable protein Application and Development in cheap protease source.
The oil crops seed is the main source of current development functionality protein.Plant physiology inspires us, and the germination process of seed is exactly that wherein various nutritional labelings (as protein, carbohydrate, fat) constantly are decomposed, and is transported to growing point with little molecular conformation, synthesizes neoblastic process again.Its germination process is exactly the active processes of various enzymes, if can in food processing process, utilize these enzymes, to be promotion, human invent the method that the amylase that utilizes in the Fructus Hordei Germinatus is produced beer, illustrate that it is feasible utilizing in the plant self contained enzyme the food industry technology.The present invention produces under this thinking promotes, and key is how to make protease enter the state of activation.
On the other hand, the raw material that can be used for preparing vegetable protein at present generally is the low temperature dregs of rice, or directly use oilseed, the high temperature dregs of rice after the high temperature oil expression then are not used (its important use is a feed), and method of the present invention can activate the enzyme of inactivating protein in the high temperature dregs of rice, has therefore developed the value of the high temperature dregs of rice.
Summary of the invention
The inventor is under the situation of above-mentioned prior art, through concentrating on studies, the proteolytic enzyme of finding oil crops self can be activated under proper condition, thereby proposed to activate the method for these proteolytic enzymes, product after the activation can replace the protease preparation that uses in the present industrial production, is used for the hydrolysis of vegetable protein.
On the basis of the above, the inventor has also proposed a kind of method of hydrolyzed vegetable protein of complete different traditional handicrafts, promptly, utilize the protease of oil crops self to realize the hydrolysis of albumen in the raw material, especially proposed to activate and utilize the method for protease in the oil crops seed, reached the reduction proteolysis and use expensive that protease preparation brought.
The present invention also provides and has utilized the resulting plant protein products of said hydrolyzed method.
According to a first aspect of the invention, proposed to activate the method for proteolytic enzyme in the oil crops seed, it comprises in the slurries of oil crops seed and to add slurries that proteolytic enzyme wherein has been activated in advance as activator, keep stirring under 6.5～8.5,30～65 ℃ of temperature of control pH value.
Method of the present invention comprises applicable to various oil crops seeds: the seed kind of soybean, peanut, rapeseed, sunflower seed, cotton seed, sesame, almond, walnut kernel etc.
Method of the present invention also is applicable to the grouts of above-mentioned raw materials seed after oil expression, comprises the common low temperature dregs of rice and the present high temperature dregs of rice mainly as feed; Method of the present invention also is applicable to the transgene product of these raw materials, for example the enzymolysis of raw material such as non-transgenic and genetically engineered soybean.
According to Activiation method of the present invention, at first need to obtain being used to start " activator " of enzyme digestion reaction, wherein contained the proteolytic enzyme that is activated through any feasible method, under the effect of this activator, the proteolytic enzyme in the raw material also can be activated.In case the protease in the raw material is activated, just can be directly be used for activating protease in the next group raw material as new activator, so handed on, the protease that utilizes raw material self to be had just can be realized the hydrolysis of protein, also just no longer need to use the protease preparation that uses in the present industrial production in addition, can reduce production costs.
The preferred scheme according to the present invention, activator can obtain by three kinds of modes:
The exogenous protease activation method;
Self activation method;
Utilize proteinase activated other oil crops protease of any oil crops that has been activated, that is, and the intersection activation method.
1, utilizes exogenous protease, that is, utilize the commercially available protein enzyme preparation to activate the method for protease in oil crops seed and the grouts.The preferred Activiation method of the present invention further comprises the described activator of preparation earlier: (the desired temperature of different enzyme preparations is different with the pH value under suitable activation condition to use protease preparation, only need requirement operation according to enzyme preparation product, or test gets final product a little) keep stirring, the pH value of observing response liquid, add the pH value that basic formulations is kept reactant liquor when beginning to descend, promptly obtain activator after 0.5～1.0 hour.
According to this method for optimizing, with raw material soaking 0～60hr such as soybean, peanut, rapeseed, sunflower seed benevolence, cotton seed, walnut kernel, sesame, almonds, add the water mill pulp, or the grouts after the above-mentioned crop oil expression directly add the water mill pulp, in being arranged, the container of automatic stirring adds a kind of or several protease preparations, for example plant rennet: papain, bromelain; Animal protease; Pepsin, trypsase; Various microbial proteases: subtilopeptidase A 1398 etc., under its optimum pH and temperature, react, can use suitable alkaline agent in the course of reaction, be preferably hydroxide or carbonate or the citrate of Na, K, Ca, Mg, keep the pH value stabilization of reactant liquor.Treat that the pH value begins decline back 0.5～1.0hr (protease in its crop seed just is activated), promptly can be used for subsequent operation.
The slurries that activate through said method can be used as activator adding next group raw slurry, and addition is generally 1/15～1/5 (weight) of next group raw slurry.That is, only use exogenous enzymes as " activator " in the hydrolysis of first raw material, the every batch of material that takes place behind the hydrolysis that is activated all can be used as activator, generations of handing on then.
2, self activation method: raw slurry at 30～65 ℃, was stirred under the condition of pH value 6.5～8.5 2～6 hours, and protease wherein can be activated, and obtains activator, during the pH value keep by adding alkaline agent.
According to this method for optimizing, oil crops seed (soak or do not soak) adds water mill pulp thing and (soaks 0～60hr, or until germinateing), the grouts that low temperature leaches legal system oil can directly add water mill pulp thing, keep 30～65 ℃ in the container of automatic stirring is arranged, and do not stop to stir, when treating that its pH value begins to descend, constantly add alkaline agent (for example Na, K, Ca, Mg alkali lye or carbonate or citrate), keep between the pH value 6.5～8.5, the about 2～6hr of the protease in the raw material begins activation.Can proceed hydrolysis this moment, or add in the follow-up raw material slurry as activator, and same the realization transmitted.
3, mutual (intersection) activation method, that is, the raw material of described activator and follow-up raw material are variety classeses.
Use exogenous enzymes to activate or automatic activation method, the slurries of any oil crops that activated or this activation slurries are removed the supernatant of telling behind the protein by acid precipitation method can be as activator, activate the protease in any other oil crops, for example Huo Hua soybean slurry or its supernatant can activate the protease in the slurries of crop seed such as peanut, rapeseed, cottonseed, sunflower and grouts, also can activate the protease of soybean, vegetable seed etc. after the same peanut slurries activation.When being appreciated that the protease of the proteinase activated of each crop and other crops of hydrolysis, speed can be different, and the effect terminal point is also different.Therefore can select the most desirable hydrolysis rate and hydrolysis terminal point by collocation.
Use comprises the activator conduct " startup source " that any method of above three kinds of Activiation methods obtains, adopt the method for similar fermentation inoculation then, the material that each batch activated inserts the next group material as " kind liquid ", activate the protease in the next group material again, production can stably go on.And except that starting activating reaction than slow, later every batch reaction speed is very fast and stable.The addition of " kind liquid " is generally 1/15～1/5 of next group raw slurry, in large-scale production, each batch raw material since kind, storage period difference and enzyme activity is slightly different with activationary time, so can adjust the reaction time by the activator ratio of adjusting every batch of adding when changing raw material, keep stable logistics speed, this is easily to those skilled in the art.
Under acid, alkali or enzyme effect, during protein generation hydrolysis, show as a large amount of insoluble proteins and change soluble protein into.The inventor has passed through a large amount of experiments, discovery is operated according to said method of the present invention, and the protein extracting ratio of raw material obviously improves, and proves that thus the protease of raw material self is activated, realized the hydrolysis of protein, concrete experimental technique and result can describe referring to the experimental example 1 of back.When preferable methods was carried out proteolysis according to the present invention, the addition of described activator was 1/15～1/5 of a raw material slurry, and the time that protease needs in the activation raw material is approximately 1～1.5 hour.
Inventor's experimental result shows that also effect was identical when the inventive method was applied to common soybeans with genetically engineered soybean, referring to experimental example 4.The enzyme auto-activation time of genetically engineered soybean is shorter than the non-transgenic soybean slightly, and other raw materials are because of reasons such as place of production kinds, and enzyme activity also has difference slightly.
Inventor's experiment also shows, by adding the method for " activator " activator protein enzyme, except the low temperature dregs of rice, be considered at present and can not also can be activated as the protease in the high temperature dregs of rice of vegetable protein raw materials for production, realize the hydrolysis of protein, referring to experimental example 2 and experimental example 5.
The present invention successfully realizes activating the protease of oil crops self, means that implementing in the enzymolysis process to vegetable protein be no longer to need to use expensive protease preparation at least, thereby can reduce the production cost of plant protein products greatly.Simultaneously, the inventor also finds, is hydrolyzed by activating oneself protease, and protein extracting ratio obviously improves, and " beany flavor " that make us unhappy in the soybean disappears thereupon; The original fragrance of seed, pungent smells such as sesame, peanut, rapeseed reduce along with hydrolysis or complete obiteration.
So, according to another aspect of the present invention, proposed to utilize said method to activate the proteolytic enzyme of raw material self, realize the method for proteolysis.Promptly, the invention provides a kind of method of hydrolyzed vegetable protein, with oil crops seed kind is raw material, utilize foregoing method, at first activate the contained protease of raw material self, make it to become " activator " in the raw material hydrolysis, add in first pending raw material slurry, under constantly stirring, keep pH value 6.5～8.5,30～65 ℃ of temperature, when treating that the pH value begins to descend, add hydroxide or the alkaline agents such as carbonate or citrate of Na, K, Ca, Mg, keep the pH value at optimum range.After finishing the enzyme digestion reaction of vegetable protein, slurry after this hydrolysis of reserve part is as " activator " of the hydrolysis of next group raw material, new a collection of slurries are kept pH value 6.5～8.5 equally under constantly stirring, 30～65 ℃ of temperature, when treating that the pH value begins to descend, add hydroxide or the alkaline agents such as carbonate or citrate of Na, K, Ca, Mg, keep the pH value at optimum range.Hydrolysis can be kept 1.5～60hr according to processing purpose, and behind the reaction terminating, 1/15～1/5 amount that leaves is as " activator ", and an additional new batch materials continues hydrolysis.The material that all the other hydrolysis are finished can enter subsequent processing, and products such as protein, polypeptide are extracted in processing.The reaction that is hydrolyzed is so repeatedly handed on hydrolytic process, can keep continuous production.
The method according to this invention is extracted protein from low temperature soy meal, compare with conventional method, extraction rate of protein can improve at least 8～12 percentage points, and reason is to utilize method hydrolysis of the present invention can make water-insoluble protein original in the soybean be hydrolyzed into soluble protein.
The inventive method does not have specific (special) requirements to the quality of water, need not soften processing yet.
The front is mentioned, and result of the test finds, utilizes the method for the present invention reaction that is hydrolyzed, and 1. can remove the original smell of seed; 2. improve extraction rate of protein more than 8～12 percentage points; 3. can reduce material viscosity, help later screenings separation circuit, simplify separation equipment, improve the concentration after material concentrates.By the said hydrolyzed reaction, changed many physico-chemical properties of protein, particularly can generate corresponding polypeptide, thereby the method for producing the vegetable protein polypeptide cheaply is provided.It should be noted that the polypeptide that obtains through the inventive method obviously reduces even eliminated the bitter taste that conventional method causes, processing produces many-sided useful influence to phytoprotein.
So the method for hydrolysis that utilizes the present invention to propose can prepare had or the still untapped plant protein products that arrives at present, for example the vegetable protein polypeptide of no bitter taste of preparation or low bitter taste.
On the basis of above-mentioned technology, the inventor also finds to utilize method for hydrolysis of the present invention to produce to have good solubility, better the instant soybeen milk powder and the soymilk series products of local flavor and mouthfeel.
Instant soybeen milk powder of the present invention, basically be major ingredient with the soybean, make slurry behind its defibrination issue unboiled water and separate, the soya-bean milk after the hydrolysis go out enzyme and spray-drying in the effect of soybean self institute soybean protein-containing enzyme, wherein, described hydrolysis is to utilize the method for aforementioned hydrolyzed vegetable protein to realize.When utilizing method of the present invention to make instant soybeen milk powder, after hydrolysis is finished, keeping an amount of beans sticks with paste in the adding next group material as " activator ", as previously mentioned, the amount of " activator " can be 1/15～1/5 (is benchmark with the slurry behind the defibrination) of raw material, can be preferably 1/10 of material quantity.The hydrolysis time of every batch of raw material is stuck with paste separation to the beans of having finished hydrolysis and is removed slag preferably at 1～2.5 hour, and the soya-bean milk pH value that filtration is obtained is reduced to 7.5～5.6 scopes, heats the enzyme that goes out immediately.This goes out enzyme process except the various harmful enzymes of deactivation, also comprises the protease that deactivation has activated.The enzyme condition of going out can be boiled 5 minutes for normal pressure; Also can high pressure go out 121 ℃ of enzymes, 30 seconds.Spray-drying can concentrate spray-drying more earlier when needing then.Spray-dired condition is identical with conventional conditions.Concentration operation can adopt the usual method vacuum to concentrate.
In said process, find that the bean dregs of hydrolysis are loose dry and comfortable, filter easily.Adopt this method for hydrolysis, the solid concentration of concentrate can reach more than 30%, helps improving spray-drying efficient like this, and can improve the unit weight and the free-running property of finished product.These prior art that all is institutes are inaccessiable.
The method according to this invention, degree of hydrolysis has tangible influence to the dissolubility of soymilk powder, selects suitable degree of hydrolysis, and soymilk powder almost can reach the solute effect of milk powder.That often adopts in this dissolving truly and the soymilk powder production at present reaches the difference that visual " dissolving " has essence by sugaring raising decentralization in water.And the bitter taste that uses the soybean protein enzyme not exist the hydrolysis of exogenous enzymes preparation to be caused.Degree of hydrolysis can be controlled by alkali consumption, if in NaOH, alkali consumption should be controlled to be in 0～3.0% scope of soybean weight, all can reach and improve the deliquescent purpose of soymilk powder.Generally speaking, the degree of hydrolysis height, alkali consumption is big, and dissolubility is good, content of peptides height in the product, but product special flavour, mouthfeel can be different.
When peanut cream that use is activated joins in the soybean material slurry as " activator ", or when comprising part peanut protein enzyme in " activator " that use, can obtain the unique peanut bean milk powder of local flavor.
Except that production had the soymilk powder of good instant capacity, the inventor also found to utilize activator protein enzyme of the present invention and hydrolyzed vegetable protein technology can produce the soya milk product of deodorization effect excellence.That is, it is major ingredient with the soybean that the present invention also provides a kind of, utilize said method to the soybean slurry be hydrolyzed obtain take off raw meat soymilk.Utilize method of the present invention to produce in the process of soymilk, adding 6～8 times of water mills after the soybean suction is saturated becomes beans to stick with paste, add " activator " that aforementioned any method is made in 1/15～1/5 ratio, 30～65 ℃, pH value 6.5～8.5, and react under the intensively stirred condition, finished reaction in about 45～90 minutes, alkali consumption is about 0.5～1.0% of soybean weight.After reaction is finished, stay 1/15～1/5 in the retort, preferred 1/10, as " activator " of criticizing raw material down, all the other are processed into and take off the raw meat soya milk product through go out enzyme (95～100 ℃, 5 minutes or 121 ℃, 30 seconds), homogeneous, seasoning etc. of heating.The purpose of this enzyme that goes out is urinated except deactivation harmful enzymes such as enzyme, antitrypsin, the protease of also wanting deactivation to activate, otherwise excessively hydrolysis also can produce a series of negative interactions.
Said process can adopt the soya-bean milk that has separated after removing slag to be hydrolyzed, and also can directly adopt beans to stick with paste first hydrolysis, separates then and removes slag.
Soybean drink market also has a kind of soy yogurt that gets consumer reception very much, but the smell of a kind of similar bean curd fermentation that it had also makes the part consumer be difficult to accept, and this has become the biggest obstacle in developing soy yogurt market.Utilize deodorization soya-bean milk that the present invention makes according to the conventional method inoculation fermentation, the soy yogurt that obtains has not then had this peculiar smell, makes the local flavor of soy yogurt and mouthfeel all be greatly improved.
Can see that in sum key of the present invention is to have proposed and the diverse vegetable protein processing thought of prior art,, manages to activate the contained proteolytic enzyme of raw material self that is, make raw material issue unboiled water then and separate reaction in the effect of this oneself protease.Compare with use with the record of prior art, the present invention has stood in the physiological property that a brand-new angle is studied oil crops, and successfully this result is applied to the deep processing of vegetable protein, not only saved a large amount of enzyme preparations, conventional process flow and corollary equipment have also been simplified greatly, thereby reduced investment and production cost, dissolubility that has solved removing soybean smell, soymilk powder simultaneously etc. adopts the traditional insoluble difficult problem of processing thought for many years.So the inventor be sure of that the theoretical research of research and processing, the especially soybean that will give vegetable protein is promoted in enforcement of the present invention and wide new space has been opened up in the production of product.
Specific embodiments
Below describe implementation process of the present invention and beyond thought many-sided positive effect of being brought in detail by concrete test and embodiment; be intended to help the reader to understand better and understand essence of the present invention and the novelty place; but but can not constitute any qualification to practical range of the present invention; in the instruction of this specification with any modification and the change made under inspiring, all should belong to the protection domain of claims of the present invention for the personage who knows art technology to technical solution of the present invention.
Illustrate: in the experimental example of back of the present invention,
Solid content in " solid content recovery rate "=slurry/raw material total amount * 100%;
Total protein content in protein content/raw material in " extraction rate of protein "=slurry * 100%
(the extraction rate of protein is here only represented the extraction effect under the manufacturing condition, does not represent the NSI value of soybean)
Experimental example 1.
The 100g soybean is saturated and become the beans paste with the 800ml water mill through soaking suction.(600,000 unit of activity/g), do not stop to stir in 42 ℃ of insulations constantly drip with 0.1N NaOH solution, keep its pH value stabilization in 7.0～7.2 to add the 0.1g papain.Cessation reaction when consumption alkali lye reaches 50ml.Get in the same beans paste of 100ml adding next batch, react by similarity condition.After this all get beans paste conducts " inoculation liquid " the adding next group that the 100ml reaction is finished at every turn, carry out 12 batches of transfer reactions altogether.
Beans behind every batch reaction are stuck with paste and are removed bean dregs respectively, measure protein and the solid content isolated in the soya-bean milk.With contrasting without hydrolysis.Result such as table 1:
Table 1
Reaction batch Enzyme dosage Alkali consumption The solid content recovery rate Extraction rate of protein
Control group 0 0 55.2％ 65.7％
First 0.1g 50ml 64.7％ 82.4％
Second batch 10 -2g 50ml 65.2％ 84.1％
The 3rd batch 10 -3g 50ml 66.1％ 86.0％
… … … … …
The 9th batch 10 -9g 50ml 65.6％ 85.3％
… … … … …
The 12 batch 10 -12g 50ml 64.7％ 85.9％
From the result of last table as can be seen: when reaction proceeded to the 12 batch, solid content and extraction rate of protein all reduced, and illustrate that enzyme activity is stable, and the papain that adds only remained 10 -12G can not keep same enzyme activity again, infers thus, must be that the protease that exists in the soybean is activated and the effect that produces.
Protease activates the influence of hydrolysis to extraction rate of protein automatically in experimental example 2. low temperature soy meals
The 100g low temperature soy meal adds 9 times of weight running water and wears into the beans paste, gets respectively with amount beans paste and does following processing:
1,, do not stop to stir in 50 ℃ of insulations;
2, adjust pH to 8.0,50 ℃ of insulations do not stop to stir;
3, do not stop to stir in 50 ℃ of insulations, when the pH value begins to descend, drip 0.1N NaOH solution, keep pH value 7.0～7.2, stopped to react in about 4.5 hours.
Above three groups of samples are separated after by the same time remove slag, measure under extraction rate of protein (in the extract in protein/dregs of beans protein) and pH value 4.5 conditions soluble protein (Kjeldahl mensuration) in the supernatant behind the heavy albumen of acid respectively, the results are shown in Table 2:
Table 2
Processing method Extraction rate of protein Soluble protein under the PH4.5
1, the neutral extraction 52.4％ 10.5％
2, alkalescence is extracted 72.1％ 11.3％
3, the automatic hydrolysis method is extracted 86.8％ 30.4％
Can see that from table 2 extraction rate of protein of automatic hydrolysis method is far above other two kinds of methods, and soluble protein also obviously improves under pH value 4.5 conditions, increases the soluble polypeptide that part generates for hydrolysis.
The automatic hydrolysis of experimental example 3. peanuts and the Hydrolyze method experiment of going down to posterity
The 100g peanut soaks a night and the 850ml running water is worn into pasty state jointly.Be incubated 50 ℃ and do not stop to stir, the pH value begins to descend after 4 hours, with the neutralization of 0.1N NaOH solution, keeps pH value 7.0～7.5, cessation reaction when consumption alkali lye reached 50ml through 1.5 hours again.Get 100ml and add in the same peanut cream of next batch, under same temperature, continue reaction.So transmit 5 batches, measure extraction rate of protein respectively and (extract protein/peanut, W/W).
Table 3
Extraction conditions Reaction time (consuming 50 milliliters of alkali lye) Extraction rate of protein PH value 4.5 soluble proteins
Hydrolysis is not extracted 24.2％ 2.8％
Automatic hydrolysis 5hr 30min 28.6％ 7.4％
The second generation 2hr 10min 27.9％ 7.5％
The third generation 1hr 33min 28.1％ 7.2％
The 4th generation 1hr 34min 28.3％ 7.3％
The 5th generation 1hr 40min 27.7％ 7.1％
As can be seen from Table 3, the proteolytic enzyme in the peanut can be activated automatically, and can transmit the protease that activates in the next group material, obviously improves through the hydrolysis extraction rate of protein.After acid was heavy, soluble protein obviously improved, and has promptly generated the peanut polypeptide.
The protease of experimental example 4. genetically engineered soybeans activates automatically and goes down to posterity activation
U.S.'s import genetically engineered soybean directly pulverized and by 60 mesh sieves.Get 100 these bean powderes of gram and 900 ml tap waters and be mixed into suspension, transfer pH to 7.0～7.5, in 50～60 ℃ of water-baths, constantly stir with 0.1N NaOH.The pH value begins to descend behind the 2hr 30min, continues to stop when reaction 1hr 45min consumption alkali lye reaches 30ml, gets this emulsion of 100ml and adds in second part of same bean powder suspension, handles once more by above-mentioned condition, stops reaction when 2hr 15min alkali consumption reaches 30ml.
Two duplicate samples are all in time separated and are removed slag, and measure solid content and extraction rate of protein, and compare by well-established law bubble beans, defibrination, the recovery rate that separates.Calculate result such as table 4 with gross protein in solid content/soybean weight, the extraction protein/soybean:
Table 4
The solid content recovery rate Extraction rate of protein
Well-established law 55.9％ 71.3％
First generation hydrolysis 66.3％ 8 3.7％
Second generation hydrolysis 65.7％ 82.9％
Well-established law is hydrolysis not 41.5％ 48.8％
The presentation of results genetically engineered soybean protease of table 4 also can be activated automatically, and can continue the activation of going down to posterity, and also can reach the purpose of activator protein enzyme without the bean powder that soaks, directly pulverize.
Again the well-established law preparation is concentrated by vacuum with first generation transgenosis hydrolysis soya-bean milk and measures its solid concentration and viscosity, its result such as table 5 respectively:
Table 5
Solid concentration Viscosity (25 ℃)
The well-established law soya-bean milk 15.0％ 147.1cp
First generation hydrolysis soya-bean milk 21.1％ 125.8cp
Experimental example 5. high temperature dregs of beans and the hydrolysis of boiling the enzyme beans paste that goes out
100g high temperature defatted soybean meal is stuck with paste through soaking the softening beans of regrinding into; Get the 100g soybean again through soaking 8hr, after wearing into the beans paste and boiling 5min, be cooled to normal temperature.Two duplicate samples all transfer pH to about 7.0, do not stop to stir in 55 ℃ of water-baths, observe the variation of pH value.Behind the 8hr, both pH values all do not change (therebetween supplementing water to keep original concentration) at any time, and each takes out the soya-bean milk liquid that 100ml removes slag through separation, measures its solid content and extraction rate of protein.And then add same concentration that 100ml has been activated respectively " activator ", press similarity condition and continue reaction.The pH value begins to descend very soon, constantly adds 4%Na 2CO 3Solution is kept pH and is stable at 7.5, and each stops reaction when consuming alkali lye 50ml, remove the gred to such an extent that soya-bean milk liquid is measured solid content and extraction rate of protein respectively.Result such as table 6:
Table 6
The solid content recovery rate Extraction rate of protein
The high temperature dregs of beans Not hydrolysis 32.6％ 34.5％
Hydrolysis 48.8％ 51.2％
Boiling the enzyme beans that go out sticks with paste Not hydrolysis 58.4％ 75.2％
Hydrolysis 67.1％ 87.0％
This test explanation high temperature dregs of beans does not have proteinase activity with the beans paste that heating was boiled, and is difficult to automatic activation, but still can be activated after adding the soybean protein enzyme " kind liquid " that has activated.But the high temperature dregs of beans is compared (when consuming alkali equally) with low temperature soy meal, and solid content and extraction rate of protein all obviously reduce; Boil beans and stick with paste recovery rate variation not obvious (comparing) with table 1.
Protease activates phenomenon automatically in the experimental example 6 various oil crops seeds
Get polly seed nucleole, almond, shelled sesame, rapeseed (juncea), each 100g of walnut kernel and add the 900ml running water respectively and smash into thin pasty state, all become white emulsion with tissue mashing machine.And to adjust pH with 0.1N NaOH be about 7.5, insulation and stirring in 50 ℃ of water-baths, and when the pH value descended, dripping alkali liquid was kept pH value 7.5.When emulsion reduced because of water evaporates, supplementing water was kept the volume of feed liquid.Record pH value begins fall time, the pH value stops to change institute's time-consuming and alkali consumption, observe phenomena.As table 7:
Table 7
Sesame Almond Sunflower seed Rapeseed Walnut kernel
Hydrolysis time 13.5hr 18hr 16hr 4.5hr 10hr
Alkali consumption 23.6ml 4.0ml 24.0ml 15.5ml 12.0ml
In the table: hydrolysis time=drop to the time that pH stops to change from the pH value.
More than the enzyme of several crop seeds all have vigor, and can activate automatically, but active, hydrolysis speed is far below soybean and peanut.After the common phenomenon of another one was hydrolysis, the original fragrance of seed, pungent smell obviously reduced even complete obiteration.And sunflower seeds is through hydrolysis, and clear liquid shows the blackish green of chlorogenic acid behind the protein precipitation.In the above several oilseeds of test explanation the protease that can be activated is arranged all.
The soymilk powder that experimental example 7 the inventive method are produced and the dissolubility check experiment of commercially available soymilk powder
Get soymilk powder (embodiment 6) and each 10.0g of commercially available sample that the inventive method is produced, add 80ml, 50 ℃ of warm water stirrings, and in 50 ℃ of water-baths, be incubated 10min, through centrifugal treating (3000rpm, 10min), discard upper solution, sediment and centrifuge tube are dried to constant weight (105 ℃), calculate insoluble matter and account for the sample ratio, relatively each sample dissolubility sees Table 8.
Sugar content≤60% among commercially available prod A in the table and the B, and milk constituents (packing indicates) is arranged.Two samples of the present invention (hydrolysis with not hydrolysis soya-bean milk powder) are soluble component such as sugaring and milk not.
Table 8
Protein content in soya-bean milk (milk) powder Insoluble matter
Hydrolysis soya-bean milk powder not 46.3％ 92.3％
Hydrolyze method soya-bean milk powder 46.3％ 2.1％
Commercially available soymilk powder A 〉=16% (packing indicates) 2.0％
Commercially available soymilk powder B 〉=16% (packing indicates) 33.1％
One night of 2Kg soybeans soaking is got in the soymilk beany flavor compare test of experimental example 8 automatic activator protein enzyme hydrolysis method manufacturings, becomes beans to stick with paste with the 16Kg running water with the emery wheel barreling again, is warmed up to 55 ℃ and stir with electric blender.Take by weighing 12g NaOH and 10g Ca (OH) 2Be made into suspension with 400ml water, and transfer beans to stick with paste pH value to 7.5 with this suspension.After the pH value begins to descend, it is constant substantially that continuation adding mixed alkali liquor is kept the pH value, treat that alkali lye adds the back and the pH value was reduced at 7.0 o'clock, separate with compact centrifuge immediately and remove slag, the gained soya-bean milk boils 5min immediately, obtain soya-bean milk 18Kg (protein concentration 3.52%), add sucrose 450g and carry out homogeneous processing (20/5MPa).
In addition, add the same defibrination of 14Kg running water, directly isolate soya-bean milk 15.5Kg (protein concentration 3.45%), add 775g sucrose, boil the same homogeneous in back and handle with one night of 2Kg soybeans soaking.These slurries are sample in contrast.
These two batches slurry sugarinesses are similar.
Buy certain brand soymilk (for the import production line is produced) from the supermarket, technical process is: raw meat, protein content 〉=3.5% are taken off in decortication, semidry method defibrination, vacuum flashing.
The trial test method is adopted in the beany flavor contrast, participant 12 people (each 6 people of men and women).Result such as table 9:
Table 9
Beany flavor Mouthfeel
No beans raw meat sensation Little sense beany flavor Beany flavor is obvious Beany flavor is strong Obvious granular sensation is arranged Granular sensation slightly Lubricious exquisiteness
Common soya-bean milk 0 0 2 10 8 4 0
Commercially available soymilk 9 3 0 0 0 9 3
Hydrolysis soymilk 10 2 0 0 0 1 11
Embodiment 1
The 1Kg soybean is soaked (temperature is below 60 ℃, and time 4～60hr can make up arbitrarily) with running water, after suction reaches capacity, add 6～8Kg water mill and become beans to stick with paste, add bromelain by carrying out enzyme digestion reaction under its optimum pH and the optimum temperature condition, and add by 2%NaOH and 2%Ca (OH) 2The mixed liquor of forming is kept the pH value stabilization.When alkali consumption reaches soybean weight 0.5～1.0%, cessation reaction, the zymoexciter that this batch materials is stuck with paste as the next group beans.
Said process also can use any other protease preparation, for example trypsase, papain, 1398 etc.
The 10Kg soybean is through above-mentioned similarity condition immersion and wear into the beans paste, in being arranged, the container of electric mixer is warming up to 30～65 ℃, make nertralizer with same mixed alkali liquor, transferring beans to stick with paste the pH value is 6.5～8.5, add first beans pastes (activator) that protease hydrolytic is crossed again, start agitator and carry out enzyme digestion reaction, after the pH value begins to descend, add above-mentioned alkali lye, it is constant to keep the pH value, according to the processing purpose difference, can control enzyme digestion reaction time 1～30hr, behind the reaction terminating, get the new beans paste of 1/10 this batch beans paste adding next group and proceed same reaction.So can constantly continue, in industrial production, can form continuous production.Except that first used enzyme preparation, later every batch of raw material all can use the enzyme digestion reaction liquid of last consignment of to make activator.
Through so beans paste viscosity reduction of enzyme digestion reaction, being convenient to screenings separates, application generic centrifuge, seperator can first separation go out loose, dry and comfortable bean dregs, bean dregs moisture content 80%, the viscosity of soya-bean milk reduces greatly, can be concentrated into solid content and reach concentration and reach 18～26% and still keep fine flowability, viscosity is less than 200cp.And the common soya-bean milk solid concentration of reference substance reaches 14～16% o'clock viscosity is hundreds of cp.The method can also improve solid content and extraction rate of protein, solid content recovery rate (accounting for soybean) can reach for 58～63% (to be 48～52% in the same old way), extraction rate of protein (accounting for soybean) can reach for 30～36% (to be 24～26% in the same old way), and a series of variations have taken place in the soybean protein character of being extracted, and always extract 25～30% of protein and are converted into soluble protein under the acid condition.
Embodiment 2
The 1Kg soybean becomes beans to stick with paste (dried soybean mill is stuck with paste fineness and should be reached 60～100 orders) through soaking (soaking conditions is with embodiment 1) or directly adding water mill without immersion, transferring beans to stick with paste the pH value earlier is in 6.5～8.5 scopes, under electronic stirring, keep 30～65 ℃ of temperature, carry out automatic enzyme digestion reaction.Behind 2.5～6hr, can be observed beans paste pH value and begin to descend, use the alkali lye among the embodiment 1 to make nertralizer, drip and keep the pH value stabilization.Stop reaction 1.0% the time approximately when what alkali consumption reached soybean weight.
Get the 10Kg soybean through soaking or wearing into beans without immersion and stick with paste, the beans paste that adds above-mentioned enzyme digestion reaction carries out enzyme digestion reaction by first similarity condition.According to enzymolysis purpose differential responses time control 1～30hr.Get following batch of beans paste of 1/10 beans paste adding then and react equally, so can successively go on.Except that first reacting activation phase is about 2～6hr, all about 1hr of later every batch materials active period.
It is all identical with embodiment 1 that the beans of the method enzymolysis are stuck with paste its separating effect, recovery rate.
Stick with paste and can not heat since the second batch materials beans, cool to 30～65 ℃ again after also can boiling, enzyme wherein can be activated again.Enzyme digestion reaction can carry out in beans are stuck with paste, and also can carry out in the soya-bean milk after separation.Resulting soya-bean milk has lost original beans fishy taste, and generation fragrance and sweet taste, become and take off raw meat soymilk, can obtain the good soymilk powder of dissolubility (this soymilk powder adds compositions such as milk and sugar) through concentrating with spray-drying again, its dissolubility can reach the level that contains sugared soymilk powder in the market.
Embodiment 3
Shelling and the peanut that takes off scarlet become pasty state through soaking or adding water mill without immersion, carry out same enzyme digestion reaction respectively by the method for embodiment 1,2, can obtain effect same.Course of reaction, temperature, pH value are all identical with embodiment 1,2.
Also available soybean paste that has activated of the reaction of peanut cream or the soya-bean milk after the separation are made " activator ", and reaction effect and reaction speed are all unaffected.
If activate soybean with the peanut cream that has activated, then soybean enzyme digestion reaction speed is faster.
Enzyme digestion reaction in the peanut cream, when proceeding to the pH value and no longer descending, adjust pH to 4.5, and centrifugation separablely go out grease, the heavy albumen of acid and soluble protein, that is and, the method can form the method for once extracting three Main Ingredients and Appearances.
Embodiment 4
Low temperature leaches low temperature soy meal or the peanut meal that legal system oil is produced, and adds water mill and becomes to stick with paste, and presses the reaction of embodiment 1,2 similarity conditions respectively: 30～65 ℃ of temperature, and pH value 6.5～8.5 stirs auto-activation time 3.5～6hr incessantly; Activator soak time 0.5～1hr.
Low temperature soy meal, peanut meal are behind the inventive method enzymolysis, and extraction rate of protein improves more than 8～12 percentage points, and viscosity reduces, and concentrate the back solid concentration and reach as high as 36%, and control reaction condition and reaction time can obtain polypeptide and other catabolites.The protein isolate that the method is made has better dissolubility.
Embodiment 5
The rapeseed and the low temperature dregs of rice thereof, condition by embodiment 1,2,4 is finished same enzyme digestion reaction, and outside reaction result isolating protein recovery rate improved, the sinapine oil content that Myronate Potassium content in the vegetable seed and reaction generate reduced, pungent obviously reduces until disappearance with the prolongation in reaction time.
Embodiment 6
The 1Kg soybean with ordinary tap water be dipped to the suction saturated after, increase weight to 2.3Kg, add 6 liters of running water again, wear into beans with the small sand disc sharpener and stick with paste, change in the stainless steel cask of band stirring and attemperator, start agitator and be incubated 45～55 ℃, it is 6.84 that beans are stuck with paste initial pH value, begins to descend through 3hr 20min pH value, treats that beans paste pH value reduced at about 6.4 o'clock, separation is removed slag, obtain 7.5 liters of soya-bean milk, solid concentration 7.4% is heated to immediately and boils and keep 5min, homogeneous (20/5MPa), utilize the vacuum film evaporimeter to concentrate, solid concentration 14% is used small-sized spray drier drying (195 ℃ of EATs more in batches, 78～82 ℃ of outlet temperatures, centrifuge 12000rpm), obtain bean powder 475g, this bean powder can be dissolved in 50 ℃ of warm water fully, do not generate sediment and supernatant, microscopically has not consoluet on a small quantity fragment.
Embodiment 7
Remove red peanut 2.0Kg the soybean of running water soaking at room temperature 14hr and 300g similarity condition soak, wear into paste jointly, in the identical hydrolysis reaction device of embodiment 1, stir hydrolysis in 50 ℃ with 6 liters of running water.Material initial pH value 6.75.Through about 2hr 45min, the pH value begins to descend.With mixed alkali liquor (NaOH: Ca (OH) 2=1: 1, melting concn 5%) drips that to keep the pH value constant.When treating that alkali consumption is 10g, stop to drip alkali, continue the about 45min of reaction, the material of taking-up 90% separates immediately and removes slag, and boils the 5min enzyme that goes out.Stay 1/10 material to add a collection of same soybean peanut again in jar and stick with paste the continuation reaction, the pH value begins to descend behind the 30min, adds same mixed alkali liquor immediately and keeps the pH value stabilization, and the 10g mixed base adds the back and continues to react 30min.Get the slagging-off of 90% feed separation equally, 10% of remainder supplies the next group reaction.So process can repeat to continue.Beginning the reaction time by second batch materials is (this time can change because of raw material is different) about 1hr 20min.The slurry that each batch obtains is handled by embodiment 1 method and is obtained bean powder.This bean powder can dissolve fully with 50 ℃ of warm water.Outward appearance is similar to fresh soyabean milk, free from beany flavor, no supernatant and precipitation.
Embodiment 8
2.3Kg soak the soybean of 14hr, add 7 liters of running water and wear into the beans paste that beans are stuck with paste and adding has been activated, in 50 ℃ of reactions that under electronic stirring, are hydrolyzed, and drip with 4%NaOH solution and to keep pH value 7.5, when alkali consumption reaches 750ml, continue reaction 30min, the separation of removing slag.Obtain about 9 liters of soya-bean milk, add 10g emulsifying agent (monoglyceride, sucrose ester etc.) and mix and boil the 5min enzyme that goes out, homogeneous (20/5.0MPa) again obtains the soymilk of fine and smooth no bitter taste., spray-drying concentrated through vacuum obtains soymilk powder.This soymilk powder dissolves in cold water, warm water and hot water.Dissolving back outward appearance such as milk sample exquisiteness, visual nothing be dissolved particles come-up and hypostasis not, and microscopically is not seen particle, and mouthfeel does not have the bitter taste that the protease preparation hydrolysis is produced yet.
Embodiment 9
100g soybean soaking at room temperature 8～14hr, add the 800ml running water and wear into the beans paste, in about 42 ℃ of water-baths, be incubated, use 0.1M NaOH adjust pH to about 7.0, electronic stirring (100～250rpm), add 0.1g bromelain or papain (600,000 units/gram), drip NaOH liquid gradually and keep 7.0, approximately through 2～2.5hr, when consumption NaOH solution reaches 15～25ml, the pH value descends very fast, illustrates that the protease in the soybean is activated, and this batch beans are stuck with paste as " activator " and used.
Use the 1000g soybean through soaking (room temperature in addition, 8～14hr), add 7 liters of running water and wear into the beans paste, add the beans paste that said hydrolyzed was reacted, keep 42 ℃ and stirring, constantly drip 0.1M NaOH solution and keep pH value 7.0, approximately through 1～1.5hr, alkali consumption 150～200ml reaction stops, taking out 90% beans sticks with paste to separate and removes slag, get soya-bean milk 8500ml, its solid concentration 7.3%, protein concentration 3.4%.The gained soya-bean milk boils cooling naturally after 3 minutes.Add sucrose 210g (sucrose concentration 2.5%) dissolving back homogeneous (20～25/5.0Mpa), obtain the no fishy smell soymilk that sugariness is equivalent to 5% sucrose, hydrolysis is described after the soymilk sweetness ratio improve without hydrolysis soymilk sugariness.
Residue 1000ml beans are stuck with paste in the retort, the beans that add the same concentration that the 1Kg soybean wears into are again stuck with paste, press the similarity condition reaction once more, stop reaction through 1hr, processing such as remove slag equally, boil, stay the 1000ml beans to stick with paste as " activator " and add new beans in the retort and stick with paste, can form serialization production so repeatedly.
Embodiment 10
The 1Kg soybean is through soaking (room temperature, 8～14hr), adding the 8000ml water mill becomes beans to stick with paste, in 50 ℃ of insulations, stir with electric mixer, and drop to the pH value with NaOH solution (4% concentration) and reach 7.0, sticking with paste the pH value through the 3.5hr beans begins to descend, dropwise add NaOH liquid and make the pH value stabilization, reach 200ml through about 45 minutes consumption NaOH amounts again, stop reaction in 7.0.Take out about 9000ml beans and stick with paste the operations such as enzyme, seasoning, homogeneous of removing slag, go out, what obtain 2.5% sucrose sweetness takes off raw meat soymilk.The beans that residue 1000ml hydrolysis is crossed in the reaction vessel are stuck with paste as " activator ", add the beans paste that the 2.3Kg soaking soybean becomes with the 8000ml water mill again, continue to press the similarity condition hydrolysis, when consumption NaOH reaches 200ml, stop reaction.This batch hydrolysis 1hr consuming time.
Repeat above operation and all can reach equal effect, except that the about 4hr of first batch of reaction time consumption, about second crowd of 1hr all consuming time approximately later on, can reach deodorization effect.
Embodiment 11
Stick with paste by the beans that embodiment 9,10 methods are made, the enzyme that goes out is boiled in heating, be cooled to 55 ℃, add any a collection of beans paste conducts " activator " that 1/10 embodiment, 9,10 hydrolysis are crossed again, by the reaction that is hydrolyzed of above-mentioned similarity condition, can reach effect same: remove beany flavor, improve protein and solid yield.Shortcoming is to boil the beans that the enzyme that goes out crosses to stick with paste, and protease is long by the time of " activation " once more, extends to 2.5hr by 1hr, illustrates that the soybean protein enzyme of inactivation can be activated once more.
Embodiment 12
900g soybean and 100g take off red peanut, press the method for embodiment 10, press the similarity condition operation, what can obtain peanut fragrance takes off raw meat soymilk, other effects and embodiment 10 are identical, and just hydrolysis speed is all fast than independent use soybean with enzyme activation speed, helps shortening the production cycle.The speed that peanut fragrance weakens is slower than the speed that beany flavor is removed.Therefore can obtain the soymilk of peanut fragrance.
Embodiment 13
Which kind of bacterial classification no matter the production of present various fermented sour soybean breasts use, and add the smell that various additives all can not be removed the similar bean curd fermentation of soy yogurt, and this has become the biggest obstacle that soy yogurt is exploited market.Adopt method of the present invention can prepare the raw material soya-bean milk of soy yogurt.
The degree of hydrolysis of any adjustment hydrolysis in the method for employing embodiment 10,11,12, alkali consumption reach soybean weight 1～2% between, the pH value is reduced to 6.5～7.0 o'clock cessation reactions, heat go out enzyme and homogeneous immediately, inoculation method fermentation routinely then, peculiar smell be can remove, soy yogurt local flavor and mouthfeel improved.

Claims (24)

1, activates the method for proteolytic enzyme in the oil crops seed, it comprises in the raw slurry of oil crops and to add slurries that proteolytic enzyme wherein has been activated in advance as activator, control raw slurry pH value is kept stirring 6.5～8.5 under 30～65 ℃ of temperature.
2, the described method of claim 1, wherein, described oil crops comprise: the seed kind of soybean, peanut, rapeseed, sunflower seed, cotton seed, sesame, almond, walnut kernel; The grouts of these raw material seeds after oil expression; The transgene product of these raw materials.
3, the described method of claim 1, wherein further comprise the described activator of preparation earlier: use protease preparation under suitable activation condition, to keep stirring, add the pH value that basic formulations is kept reactant liquor when the pH value of observing response liquid begins to descend, promptly obtain activator after 0.5～1.0 hour.
4, the described method of claim 3, wherein, the protease preparation of employing comprises animal protease, plant rennet and microprotein enzyme preparation.
5, the described method of claim 1 wherein further comprises the described activator of preparation earlier: raw slurry at 30～65 ℃, was stirred under the condition of pH value 6.5～8.5 2～6 hours, during the pH value keep by the adding alkaline agent.
6, the described method of claim 1, wherein, described activator can from the different types of oil crops of raw material.
7, each described method of claim 1～6, the addition of wherein said activator is 1/15～1/5 of a raw material slurry weight, the time of activating proteolysis enzyme require in the raw material slurry is 1～1.5 hour.
8, each described method of claim 1～6, described alkaline agent is selected from hydroxide, carbonate or the citrate of sodium, potassium, calcium or magnesium.
9, the method for hydrolyzed vegetable protein, with oil crops seed kind is raw material, utilize each described method of claim 1～6, at first activate the contained proteolytic enzyme of raw material self, make raw material issue unboiled water in the effect of this proteolytic enzyme and separate reaction, hydrolytic process is kept stirring, 30～65 ℃ of temperature controls, add alkaline agent control pH value 6.5～8.5, hydrolysis time 1.5～60 hours.
10, the described method of claim 9, it comprises: utilize any protease preparation at first to make raw material slurry generation hydrolysis, when beginning to descend, the pH value of observing response liquid adds the pH value that basic formulations is kept reactant liquor, 0.5 promptly obtain activator after～1.0 hours, then this activator is added in the raw slurry for the treatment of hydrolysis in the first batch and carry out enzyme digestion reaction, after enzyme digestion reaction was finished, the slurry after this hydrolysis of reserve part was handed on hydrolytic process as the activator of next group raw material enzymolysis.
11, the described method of claim 10, wherein, the protease preparation of employing comprises animal protease, plant rennet and microprotein enzyme preparation.
12, the described method of claim 9, it comprises: with raw slurry at 30～65 ℃, stirred under the condition of pH value 6.5～8.5 2～6 hours, pH value is during this time kept by adding alkaline agent, the plant oneself protease is activated, adds in the raw material slurry for the treatment of hydrolysis in the first batch as activator with this hydrolysis material then and carry out enzyme digestion reaction, after enzyme digestion reaction is finished, slurry after this hydrolysis of reserve part is handed on hydrolytic process as the activator of next group raw material enzymolysis.
13, each described method of claim 9～12, wherein, the raw material of preparation activator slurry can be different with the raw material type that is used for hydrolysis.
14, claim 10 or 12 described methods, wherein, described alkaline agent is selected from hydroxide, carbonate or the citrate of sodium, potassium, calcium or magnesium.
15, claim 10 or 12 described methods, wherein, the addition of activator is 1/15～1/5 of a follow-up raw material slurry weight.
16, a kind of plant protein products utilizes each described method preparation of claim 9～15.
17, the described plant protein products of claim 16, it is vegetable protein polypeptide or soy protein isolate.
18, a kind of instant soybeen milk powder is to be major ingredient with the soybean, and the method for utilizing in the claim 9～15 each is to the raw material back preparation that is hydrolyzed.
19, the manufacture method of the described instant soybeen milk powder of claim 18, comprise that the slurry that makes behind the defibrination issues unboiled water in the effect of the contained soybean protein hydrolysis enzyme of soybean self and separates, described hydrolytic process is to utilize in the claim 9～15 each method to finish, the soya-bean milk after the hydrolysis go out enzyme and spray-drying.
20, the described manufacture method of claim 19, wherein, in the hydrolytic process, alkali consumption is 0 of raw material weight～3.0% o'clock, hydrolysis is finished.
21, a kind of soymilk for being major ingredient with the soybean, utilizes each method of claim 9～15 that the soybean slurry soya-bean milk that obtains that is hydrolyzed is made.
22, the manufacture method of the described soymilk of claim 21, comprise that adding 6～8 times of water mills after making soybean suction saturated becomes beans to stick with paste, issuing unboiled water in the effect of soybean self institute soybean protein-containing enzyme separates, described hydrolytic process is to utilize in the claim 9～15 each method to finish, wherein, alkali consumption is about 0.5～1.0% of soybean weight; After the soya-bean milk of reaction after finishing reserved part as activator, all the other hydrolysates heating enzyme that goes out.
23, the described manufacture method of claim 22, wherein, hydrolysis time control 45～90 minutes.
24, a kind of soy yogurt is to utilize the soymilk of claim 21 to form through inoculation fermentation.
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