CN1318603C - Method for preparing plant separated protein - Google Patents
Method for preparing plant separated protein Download PDFInfo
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- CN1318603C CN1318603C CNB2003101208352A CN200310120835A CN1318603C CN 1318603 C CN1318603 C CN 1318603C CN B2003101208352 A CNB2003101208352 A CN B2003101208352A CN 200310120835 A CN200310120835 A CN 200310120835A CN 1318603 C CN1318603 C CN 1318603C
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- Peptides Or Proteins (AREA)
Abstract
The present invention provides a method for preparing plant separated protein, which adopts a new method which is completely different from the classic production principle of plant separated protein. Proteinase (one of the dynamic force for seed budding) in a raw material is firstly activated in the present invention, and protein in the raw material is dissolved; after the protein is separated and deslagged, an enzymolysis reaction is continuous, and protein is precipitated at an isoelectric point by the acidity generated in a hydrolytic process. The method belongs to a biochemical method, acid and alkali are not used in a large quantity, and no salt pollution is generated. The production flow is simple, and the functions of solubility, etc. of the product are greatly improved. The method also provides convenience for extracting side products, such as isoflavone, oligosaccharide, etc., from whey. The present invention especially provides the continuous production technology of soy separation protein and groundnut separation protein.
Description
Technical field
The present invention relates to the deep processing field of vegetable-protein, relate in particular to and utilize the contained proteolytic enzyme of plant self, adopt biochemical process to extract the method for plant separation proteins such as making soybean and peanut as albumen stripping agent and precipitation agent.
Background technology
Vegetable-protein, especially the extraction of soybean protein isolate research starts from the forties in last century in the U.S., very perfect to the eighties, the technology that has formed with a number of major companies is the production technology of representative, and be considered to the complete sophisticated classical technology of a cover, write into the specialized textbook of the college of engineering, to such an extent as to beginning most of scientists from the eighties changes over to one after another to soybean protein and oligose, the research of physiological functions such as isoflavones has also obtained large quantities of scientific payoffss, allow people that soybean nutritional and physiological function are produced new understanding, the research center of gravity has also been left the soyabean processing field.
Soybean protein isolate is to be raw material with the defatted soybean meal, separates purification through removing the non-protein ingredient of solubility with insoluble macromolecule component, contains protein and is not less than 90% soybean protein powder.Before the present invention, the representative processes route that extracts soybean protein isolate all is an alkali extraction and acid precipitation, promptly utilize soybean protein solubility rate under alkaline condition to improve and in the method that its iso-electric point (under the acidic conditions) is settled out, produce soybean protein isolate, its theoretical basis is chemical process.Traditional technology is generally raw material (for example low temperature soy meal) is added the water mill pulping, add NaOH liquid adjust pH 9.0-11.0 (add alkali lye and be used to improve proteinic solvability), in 15-80 ℃ of stirred for several ten minutes, make the protein ingredient stripping, add acid (adding hydrochloric acid usually) protein precipitation then.Need to consume a large amount of NaOH in the alkali extraction method of this traditional technology, acid consumes a large amount of acid again and causes and produces a large amount of salt in the byproduct when heavy, for from whey liquid, extracting the difficulty in isoflavones, the oligose increase production, increase the desalter input, raise the cost greatly and cause environmental pollution.
Along with people's going deep into to the physiological function research of deep processed products such as soybean separation protein bletilla oligose, isoflavones, the Application Areas of these products is also continually developed, especially more and more accepted as the function of medicine or healthcare products by people, soybean protein isolate is as the important source material of functional foodstuff, demand also increases suddenly, and the method for exploitation high-level efficiency, low cost production protein isolate, isoflavones and oligose also just has very practical meaning aspect improving the economic and social benefits.
Summary of the invention
The inventor through exploration discovery for many years the oil crops seeds, especially supply the utilizability of proteolytic enzyme in industrial production of physiological development in soybean, the peanut and other crops, found under working condition, activate its active method within a few hours, and proposed application for a patent for invention (application number is 02157397.2).On this basis again through exploration and the effort in 1 year, discovery utilizes the activation of these proteolytic enzyme can finish stripping and the method that precipitates two important procedures automatically, promptly, not exogenously added alkali and acid, under the effect of plant oneself protease, realize proteic extraction and separate, can replace the production that traditional technology realizes soybean, peanut protein isolate product fully.
So, the invention provides a kind of novel method of producing plant separation protein, on principle, be different from the traditional chemical method fully, no longer need to use a large amount of bronsted lowry acids and bases bronsted lowries, production technique and equipment all obtain simplifying.
The present invention especially provides the production method of soybean protein isolate, peanut protein isolate, and the protein isolate that obtains is meeting or exceeding the protein isolate of produced in conventional processes in nature.
The method according to this invention, at first activate the contained proteolytic ferment of raw material self, making raw material issue unboiled water in the effect of this proteolytic ferment separates reaction and obtains protein emulsion, when the pH of this emulsion value reaches 6.5-6.0, separate slagging-off, make protein emulsion continue hydrolysis, near the pH value reaches the albumen iso-electric point, separate and collect protein precipitation and whey liquid, adjusting protein precipitation becomes neutral protein liquid, and drying becomes separated protein powder then.
According to method of the present invention, separated protein emulsion continues hydrolysis reaction under the effect of enzyme, do not add alkali control pH value this moment, near protein emulsion pH value is reduced to the albumen iso-electric point, protein precipitates in the sour environment that self hydrolysis produces, and can obtain protein isolate through centrifugation.Through transferring processes such as neutrality, sterilization, spraying drying, can obtain separated protein powder.Through felicity condition control, its solvability will improve greatly than the chemical process that it is heavy that traditional alkali is proposed acid, can reach the milk-protein level.
Aforesaid method goes for the extraction of various vegetable-proteins, and for example: the remaining grouts of oil expression of oil crops such as soybean, peanut, vegetable seed, sunflower seed or leaching back all can be used as and utilize this method to produce the raw material of separated protein powder.Albumen according to raw material is formed and content, and what relatively be suitable for suitability for industrialized production is soybean protein and Semen arachidis hypogaeae protein.Be appreciated that, in the actual production, different according to raw material and albumen kind, the iso-electric point that protein emulsion continues in the hydrolytic process is different, in the extraction for described soybean protein or Semen arachidis hypogaeae protein, general may command pH5.0-3.0 separates when preferably being controlled at 5.0-4.0, and the selected character and the purity that also depends on the protein ingredient of desire acquisition of the pH value scope when beginning to separate.
Still contain more protein in the whey liquid behind the precipitation separation albumen, can heat the Partial Protein that makes once more wherein and solidify precipitation, collect secondary sedimentation albumen, adjustment becomes neutral protein liquid (using alkali lye generally speaking), drying becomes protein powder, perhaps this secondary sedimentation albumen and protein precipitation merging is last time handled.
About activating the method for the contained proteolytic ferments of raw material self such as soybean, the contriver has had a detailed description in first to file 02157397.2, can adopt exogenous protease activation method, self activation method or activation method that intersects.The contriver introduces the present invention as a reference in full with specification sheets and claims of this application case.So based on method of the present invention, the process that activates the contained proteolytic ferments of raw material self such as soybean comprises and adopt one of following method preparation activator, utilize this activator to start behind the hydrolysis reaction of raw material then as time activator of batch raw material.
1) utilize any protease preparation to make raw material slurry generation hydrolysis under optimum conditions;
2) with raw slurry at 30-65 ℃, stirred under the pH value 6.5-8.5 condition 2-6 hour, during pH value keep by adding alkaline agent, make the contained proteolytic enzyme of raw material self be activated the back as the activator of time criticizing raw material;
3) protein emulsion that obtains after preceding a collection of raw water is separated, the perhaps direct activator of whey liquid behind its continuation hydrolysis precipitation separation albumen as following batch raw material.
Wherein, the raw material of preparation activator can be soybean, peanut, sesame, almond, walnut kernel, sunflower seed, Semen Brassicae campestris or cotton seed.As the contriver above-mentioned described in the first to file, in case the activation of the proteolytic enzyme in the realization raw material, whey liquid after every batch of hydrolysis or wherein a part just can stay the activator of doing next production cycle, so, only need before the hydrolysis of raw material in the first batch, prepare activator.Use these activator as the startup source, adopt the method " inoculation " of similar fermentation inoculation in the raw material of next batch, to be used to activate new proteolytic enzyme, start the new production cycle.
As can be seen from the above description, though method of the present invention and the traditional similar part of classical way are still to comprise two operations of stripping (protein extraction) and precipitation, the principle of its realization is diverse.Classical way is a chemical method, utilizes alkali to finish process in leaching for chaotropic agent, also further reduces the pH value to iso-electric point with the acid neutralization again, and two-step all uses the Chemicals starting material.The inventive method is a principle of having used for reference plant physiology, utilize the active substance processing oneself of plant oneself: promptly the proteolytic enzyme in the plant seed germinating growth process is applied to industrial production, make the original protease activated process that a couple of days finishes in germination process shorten to a few hours, and utilize these proteolytic enzyme to finish plant protein stripping and two processes of precipitation.
So key of the present invention is to adopt enzyme process separation and Extraction and enzyme process precipitation.Extraction process is to utilize the short solvency action in protease activated early stages such as soybean to replace the alkali in the chemical method to put forward technology, the enzyme process depositing technology then is to utilize the further hydrolysis of proteolytic enzyme that albumen is precipitated in the sour environment that self forms, and replaces the heavy technology of acid in the chemical method.The concrete implementation method according to the present invention is ground into dregs of beans earlier 100-120 purpose fine powder or adds water and directly wears into screened stock with colloidal mill.The preceding less generation foam of a kind of method; A kind of method in back adds when water grinds can produce foam, is unfavorable for operation.The present invention does not have particular requirement to water hardness, need not to use softening water, as long as meet the foodstuffs industry water supply standard.This beans are stuck with paste and are placed the enzyme activation jar extractor of holding concurrently, and pH value can be beans and sticks with paste the nature value,, need not add alkali adjusting pH value that is, and also adjust pH is no more than 8 slightly, for example 6.5-6.8.Add activator, perhaps at first make protease activated according to self activated method, for helping hydrolytic process, general control material is at 30-65 ℃, be preferably under the 35-50 ℃ of temperature and stirred 1.5-3.0 hour, treat that the pH value begins to descend or when reducing 0.2-0.3 (6.5-6.0), this moment, protein emulsion viscosity obviously reduced, and can be easy to separate slagging-off and collect protein extract.After measured, utilize the protein extracting ratio of enzyme extraction and traditional alkaline extraction method basic identical, can reach 87-92%, but the outward appearance of this protein extract such as common soya-bean milk sample oyster white, so claim protein emulsion, and the outward appearance of alkali extracting solution is the semitransparent brown yellow.Extracting solution after the slagging-off (protein emulsion) enters the Acid precipitation jar, continue to stir (100-120rpm), be actually the continuation hydrolytic process, temperature can be identical with the temperature in the enzyme activation extractor, also can different (exceeding) to guarantee the proteolytic enzyme non-inactivation, generally at 0-65 ℃, this moment, slurry pH value continued to descend, the pH value can be reduced between the 5.0-3.0 after 0.5-2.5 hour, between the preferred control 5.0-4.0, for example soybean protein can be controlled at about 4.5, and Semen arachidis hypogaeae protein can be controlled at 4.5-5.0, determines to finish precipitin reaction.
In industrial production, said process can be an intermittent type, the extraction of first material needs first activated protein enzyme to start enzymolysis, the a collection of material that activated gets final product (add-on 1/10-1/5) before adding in later every batch of new material, need at least 2 group jars to be used alternatingly like this, equipment volume is bigger, and enzyme activity all experienced initially to complete activatory process at every jar of material, and required time is longer.So enzymolysis and precipitation process adopt continuation method in the preferred embodiment of the invention, after the material in hydrolysis extractor and the precipitin reaction jar met the requirements of the pH scope, constant speed added new feed liquid when reaction mass is finished in discharge.
After enzyme activation extracts and to reach requirement, from the end of jar or the top discharge beans and stick with paste and enter separating centrifuge, and with constant speed from the top or the bottom replenish the beans that newly grind and stick with paste, enter normal following current production process.The alternate cycle of material can be controlled at 50-80 minute in this postactivated extractor, generally need about 1 hour (a promptly complete jar material is discharged fully by exchange and needed about 1 hour), and it is constant to keep activation pH value, this process can be by the control of pH value feedback two-position controller, also fixing alternate cycle is controlled by stability of flow.
Extracting solution enters the Acid precipitation jar and continues hydrolysis under the enzyme effect, after determining to finish precipitin reaction, beginning is discharged the slurries of finishing precipitin reaction with the uniform velocity limit and is entered centrifugation for the second time from jar, the new protein emulsion that obtains through zymohydrolysis extracting method is sent on the limit, thereby enter normal productive process, the slurries of discharge obtain protein precipitation and whey liquid through centrifugation.Feed liquid in the Acid precipitation jar during ordinary production alternate cycle can generally be about 1 hour at 45-120 minute, a promptly complete jar material per hour exchanges once.
In actual production,, can calculate the volume of jar to reach desirable exchange cycle (yield in unit time, retort volume and reaction time are funtcional relationship between the three) according to speed of response as adopting continuous processing.
Can see that extraction and precipitation all adopt the technology of continuous processing, compare with interrupter method, the enzyme in jar in the material is in high vigor state always, has shortened the production cycle, has reduced equipment volume.
In the above method (interrupter method or continuous processing) as adopt the method for self activation proteolytic enzyme, start-up period is longer, about altogether 4-4.5 hour of stripping and precipitation, if utilize any protease preparation to quicken the cycle of activating or use to produce the whey liquid that retains as activator, start-up period is shortened greatly according to above-mentioned.
The contriver finds in research process, it is different to adopt sedimentary albumen mud of protease method and the main difference of albumen mud that acid precipitation method obtains to be embodied in water ratio: the albumen mud water ratio of the inventive method is low, at 80-60%, and the sedimentary albumen mud of acid system water ratio is generally more than 80%.The further processing of this albumen mud is similar to traditional method, through washing, transferring neutral dispersing and dissolving (pH value 7.0-7.5), sterilization, spraying drying to obtain separated protein powder, but solubility experiment shows, the inventive method is produced the solvability of protein isolate than acid precipitation method height, but the translucent Colloidal fluid before the melt into spraying drying.
After protein precipitation mud is finished and isolated in precipitin reaction, also containing more protein in the whey liquid, mainly is the low molecular weight protein (accounting for the 10-15% that is extracted total protein) that hydrolytic process produces.The heated milk clear liquid can make this part albumen separate out, can obtain secondary albumen mud through centrifugation, again it is neutralized to the pH value and is dissolved into protein solution for 7.0-7.5, spraying drying obtains the good protein powder of solvability separately, also can sneak into the common drying of flash liberation albumen and obtain identical product.Described whey liquid also can be used as the deep processing raw material of other protein products.
Soybean protein isolate with the most generally production in more than describing is an example, and based on the consideration of protein content, another kind can top-priority raw material be exactly a peanut in the oil crops, can obtain peanut protein isolate equally.So the raw material that utilizes the inventive method to produce protein isolate preferably can be the low temperature dregs of rice, the high temperature dregs of rice or its mixture of soybean, genetically engineered soybean or peanut etc.When adopting the high temperature dregs of rice to do raw material, because the enzyme activity of himself is lower, the whey liquid that preferred use has been activated for improving the protein product yield and shortening the reaction times, preferably comprises a certain proportion of low temperature dregs of rice as the method for activator in raw material; Perhaps, use the low temperature dregs of rice, enter and to use after the successive reaction high temperature dregs of rice or high temperature, the low temperature dregs of rice to mix to use at the first batch of startup raw material that extracts with setting tank.The ratio of high temperature dregs of rice this moment in mixture can be 0-90%.
From above narration as can be seen, the present invention has changed traditional processing method, adopt biochemical method to utilize the proteolytic enzyme dissolving of soybean etc. self to extract albumen,, but have essence different with traditional method though production stage is proteins extraction and precipitate and separate.
Traditional method is that alkalescence is extracted, and generally is that raw material (dregs of beans) and water are ground, and adds NaOH liquid and be transferred to pH value and be 9-11, heat tracing in extractor (general 15-80 ℃) stirring 20-120 minute, and extraction yield can reach 85-90%.The plant oneself protease of the inventive method after utilization is activated under basic neutrality and the proper temperature is as chaotropic agent, and extraction yield can reach the same efficient of alkaline process.Though yet insulated and stirred in the tradition alkaline extraction, enzyme are not activated (according to traditional theory, not only do not advise making the enzyme activation of self, common way is the enzyme that goes out of heating earlier, to remove so-called raw meat, bitter taste).The current precipitation of protein of our times adopts HCl, H
2SO
4The pH value of adjusting the alkaline extraction emulsion makes albumen precipitation and obtains paste albumen through centrifugation to iso-electric point 4.5; The inventive method is then utilized through the isolating protein slurry of enzyme process and is let alone to continue enzyme digestion reaction, in the time of near the pH value oneself is reduced to iso-electric point again centrifugation obtain paste albumen, the H+ that the amino acid end ionization that acidity in this process produces from proteolysis discharges, do not introduce external negatively charged ion in the whey liquid, therefore also needn't be with in a large amount of alkali secondaries and whey liquid.Carry in the heavy technology of acid at the alkali of classics, the soybean protein isolate that every production is a ton consumes 60Kg caustic soda and 110Kg concentrated hydrochloric acid approximately, and in whey residual a large amount of salt and hydrochloric acid, discharging can pollute, if separate then cost is very high.
Therefore, the inventive method has been avoided a large amount of aborning use acid, alkali, has reduced corresponding operation and equipment, material and equipment cost have been reduced, reduced salt and the acid in the whey liquid, avoided secondary pollution, and reduced difficulty for further extracting oligose and isoflavones.
Method of the present invention is compared with the classical acid alkaline process, as adopts self activation method for enzyme, and the startup reaction that extractor and enzyme are heavy jar all needs certain hour, so just can enter continuous production in 3-4 hour consuming time approximately when producing beginning.If but first jar extract use in the material protease preparation or on batch produce the whey liquid that retains, extractions, sedimentary startup are reacted respectively foreshorten to 1 hour, even shorter.
Meaning of the present invention is that a physiological function of plants sexual element is used for industrial production, the further investigation because a large amount of mechanism problem wherein and reaction process are still needed, so the present invention has also opened up the frontier of the direct industrial utilization research of vegetable-protein science and technical study and biological enzyme.The inventive method is compared with the alkali extraction and acid precipitation of classics, and following advantage is arranged:
1, acid-base method is produced soybean protein isolate and will be used a large amount of soda acids, has increased production cost and equipment, and whey liquid is handled also numerous and diverse.The inventive method is not used soda acid and has been reduced cost of supplementary product and relevant device, eliminated the salt in the whey liquid, reduced pollution emissions, because there has not been exogenous salt in the whey, and then process when extracting oligose, isoflavones and exempted the desalination operation, reduce equipment and tooling cost greatly.
2, the inventive method is when having simplified whole technological process and equipment, and the productive rate of protein extracting ratio and protein precipitation all reaches traditional alkali extraction and acid precipitation level, can also more be better than the result of traditional technology by control of process condition.
3, the soybean protein isolate of producing according to the inventive method also has good solubility, can reach or near the milk-protein solvability, can reach the collosol state before the spraying drying behind the molten water.
4, by control of process condition, lower molecular weight vegetable-protein of production part simultaneously and plant polypeptide.
5, the low temperature dregs of rice of the enforcement of the inventive method after using soybean extract oil, be considered at present and can not also can be activated as the proteolytic enzyme in the high temperature dregs of rice of vegetable-protein raw materials for production and utilize, so, proteic raw material sources enlarged.
6, the present invention does not have particular requirement to production unit, now uses the manufacturing enterprise of traditional technology to simplify transformation for existing protein isolate production line, can satisfy processing requirement of the present invention.
7, the beany flavor of the soybean protein isolate of the inventive method production is significantly less than the product of traditional method.
Enzyme system in the plant seed is very complicated, and the existence of each enzyme all has the life meaning.Corresponding to existing any composition (protein, fat, starch, Mierocrystalline cellulose or the like) in the plant seed, certainly exist the enzyme that decomposes or synthesize them, otherwise seed maturation and germinating growth process just can't be carried out.The present invention and contriver openly will point out people in the first to file described content: the enzyme in the plant may directly be utilized by industrialization, at least wherein proteolytic enzyme can directly utilize, and it utilizes the result that the trade union that adds of plant protein is produced profound influence.In foodstuffs industry was produced, most complete processings were harmful object with biological enzyme all, can produce unfavorable phenomenons such as bad flavor, color and luster instability, appearance precipitation because of it, so food-processing all has the enzymatic process of going out.Thinking of the present invention be with biological enzyme for utilizing object, utilize earlier and afterwards go out it.We can say, the present invention's (comprising described in first to file) has proposed a kind of thinking of going with anti-its road of traditional theory, sum up effect front technically, when obtaining these results, the contriver also sighs with deep feeling from the deep of the heart, these results are that the contriver is subjected to the enlightenment of plant physiology, medical science and the imagination that inspires, form through the effort for many years and the countless detours of passing by.If having proved once more, it in the subject crossing process, suitably changes the thinking direction, just might find " There is a way out ", simultaneously when reading many forefathers' documents once more, find to have that the investigator also once went to this discovery fails to expect pushing open it greatly in front of the door.As seen so only one step away change be based upon the contriver happen suddenly inspiration and explore untiringly with the paying of creative work among, after the effort, successfully just in " on looking back " of chance.
Description of drawings
Fig. 1 is the process flow sheet of the preferred embodiment of the invention.
According to this technology shown in Figure 1, when raw material was low temperature soy meal, the product that obtains was a soybean isolate protein powder, contained the protein of 18-35% in the by product, can be used as feed.
Embodiment
Further describe the present invention below in conjunction with concrete scheme; embodiment wherein only is a preferred version of the present invention; be intended to help the reader to understand better and understand essence of the present invention and the novelty place; but can not constitute any qualification to practical range of the present invention; the personage who knows art technology all should fall into the protection domain of claims of the present invention in the instruction of this specification sheets with any modification and the change to embodiment of the present invention done under inspiring.
Test example 1, Enzymatic Extraction and alkaline process extract relatively
Get protein content and be 6 parts in 48.2% low temperature soy meal powder (120 order), every part 200 gram, each adds 2200 milliliters of ordinary tap water furnishing beans and sticks with paste.Wherein three parts are respectively: natural pH value (about 6.4), be transferred to pH value 9.0 and 11.0 with NaOH solution respectively, be numbered 1#, 2#, 3#, (25 ℃) stirred 20 minutes under room temperature, respectively get 400 milliliters of small-sized separating centrifuge same rotational speed of usefulness and isolate bean dregs, measure protein content (Kjeldahl determination) in the slurries, centrifugation and measure protein concentration when leftover materials are stirred to 120 minutes.Other three parts of beans are stuck with paste and also are respectively nature pH value (4#) and with NaOH solution adjust pH 6.8 (5#), pH value 8.0 (6#), be rapidly heated after 45 ℃, insulated and stirred (100rpm) and take out 400 milliliters of material centrifugations respectively at 0.5 hour, 1.0 hours, 1.5 hours, 2.0 hours in 45 ℃ of water-baths is measured protein content in the slurries respectively with quadrat method.Result such as table one.
Table one
| The numbering condition | 1# | 2# | 3# | 4# | 5# | 6# | |
| Time | 20 minutes | 3.39％ | 3.82％ | 3.99％ | |||
| 30 minutes | 3.77％ | 3.82％ | 3.82％ | ||||
| 60 minutes | 3.85％ | 3.86％ | 3.84％ | ||||
| 90 minutes | 3.90％ | 3.95％ | 3.87％ | ||||
| 120 minutes | 3.65％ | 3.90％ | 3.99％ | 3.90％ | 3.95％ | 3.87％ | |
| The PH value | Initial | 6.40 | 6.80 | 8.00 | |||
| Terminal point | 6.20 | 6.60 | 7.57 | ||||
From table one as seen: add alkali under the normal temperature and directly extract, improving the protein solubility rate with the pH value also improves, and protein concn is the highest during pH value 11.0, but consumption NaOH amount is about 2.8% of dregs of beans under this condition, can consume more acid during corresponding acid adjustment, cause cost to improve and the utility appliance increase, and contain a large amount of salt in the whey, the later stage will increase pollution and desalter and process cost.And 4#, 5#, 6# sample promptly reached maximum separately through insulated and stirred in 1.5 hours, be respectively 3.90%, 3.95% and 3.87%, similar to extraction yield under the highly basic condition of pH value 11.0, but isolated slurries are near neutral, be beneficial to and continue reaction and aftertreatment, and 5# and 6# material liquid pH value can slowly descend in reaction, the pH value 0.2-0.6 that can descend respectively can directly enter separation circuit after 120 minutes.
Preferred extraction conditions is selected to be defined as stirring 30-55 ℃ of insulation 1.0-1.5 hour in view of the above.Compare with alkali extraction method, except that protein concentration was identical, material viscosity obviously reduced, similar during to pH value 11.0, not only satisfy the protein extracting ratio requirement, and be beneficial to the screenings separation, isolated bean dregs are dry and comfortable loose, and water ratio can be reduced to 72-78% by the 82-85% of well-established law.The slurries yield improves, and has further improved the albumen yield again.If add a collection of slurries before the activatory in material, extraction time can also shorten.(annotate: this case said " pH is value naturally " is meant and does not add any alkali or acid, the pH value of material.)
Test example 2, the enzyme precipitator method and acid precipitation method are relatively
In the inventive method, do not use the alkali neutralization behind the enzyme activation, let alone the decline of pH value and continue reaction, reduce to 4.5 until the pH value, this process still needs 1.5-2 hour, and continuously rule to keep the material exchange cycle be 1 hour, the settle out difference of process and traditional technology of this albumen is to need not to add acid, utilizes the sour environment of enzymolysis process formation to make albumen precipitation.The protein precipitation yield compares with the heavy method of acid, confirms with following test.
Utilize the soya-bean milk of 1#, 2# in the test example 1,3# method (20 minutes extract at room temperature) preparation to use hydrochloric acid adjust pH to 4.5 respectively, the centrifugal protein precipitation of telling, whey liquid is heated to about 65 ℃ and produces a small amount of flocculent aggregate, separate through recentrifuge, measure residual soluble proteins in the whey liquid second time respectively.Merging protein precipitation twice, is 4.5 sour water washing 2 times with the pH value respectively, measures protein precipitation (Kjeldahl method) more respectively.
4#, 5#, the 6# sample activation soya-bean milk (protein emulsion) that also to be test example extract with quadrat method reacts to the pH value in 50 ℃ of continuation reduces to centrifugation in 4.5 o'clock respectively, heating, separation, measure protein concn in the second whey liquid, protein precipitation merges with the water washing of pH value 4.5 2 times, Kjeldahl method mensuration protein precipitation yield.
More than six samples all get 1000 milliliters of emulsions tests.
Table two
| 1# | 2# | 3# | 4# | 5# | 6# | |
| Protein concn (1) | 3.39％ | 3.82％ | 3.99％ | 3.90％ | 3.95％ | 3.87％ |
| Protein concn (2) in the second whey of the heavy back of pH value 4.5 acid | 0.59％ | 0.59％ | 0.59％ | |||
| PH value 4.5 enzyme post precipitation second whey protein concns poor (2) | 0.59％ | 0.59％ | 0.59％ | |||
| (1)-(2) | 2.80％ | 3.23％ | 3.40％ | 3.31％ | 3.36％ | 3.28％ |
| Actual protein precipitation amount (dry weight gram) | 27.1 | 31.2 | 32.6 | 32.0 | 32.4 | 31.6 |
As seen from Table 2, no matter be slurries neutral, alkaline extraction method, again through the heavy method of acid or enzyme extraction, the heavy method of enzyme, can not the protein precipitation amount be constant in the second whey liquid, the heavy method of enzyme is identical with the heavy method effect of acid, illustrates that this part soluble proteins is not subjected to extract, intermediate processing influences, and quantity is constant.Concentration difference pH value 11.0 alkali extraction methods (3#) and the two effect approximately equal of pH value 6.8 (5#) the enzyme extraction enzyme precipitator method as can be known as calculated.
Above through enzyme extraction and enzyme intermediate processing extraction protein, the protein isolate yield is similar to alkali extraction and acid precipitation.The provable enzyme solution of its effect can replace the current alkali extraction and acid precipitation of our times fully.In addition, acid is heavy to have notable difference with the heavy resulting property of protein of method of enzyme, and the protein precipitation cream water ratio that the heavy method of enzyme obtains is lower than the heavy method of acid, can reduce to 60-65%.Be under the same centrifugation condition, enzyme sinks albumen dehydration rate height.
Test example 3, continuous processing are extracted
Add 11.5 times of tap water with the low temperature soy meal 1.5Kg that contains albumen 48.2% and make the beans paste,,, be stored in room temperature (18-25 ℃) as initial material with small amount of N aOH liquid adjust pH to 6.6 with colloidal mill.
Get 1200 milliliters of these beans paste liquid and in beaker, insert 45 ℃ of water-baths, adopt electronic stirring (100rpm).Installing a siphon pipe in advance draws slurries and regulates take-off rate 20 ml/min, transfer to same speed with high-order tubualted bottle simultaneously and in beaker, add slurries, after regulating this device, make first batch of 1200 milliliters of material reactions after 1.5 hours earlier, again with per minute turnover material 20 milliliters of speed exchanges of liquid (promptly per hour exchanging a collection of), reacted beans are stuck with paste liquid, every collection is a collection of for 1200 milliliters, centrifugation is removed slag and is obtained soya-bean milk, measures protein concn and solid concentration in first, fourth, seven, ten batch of emulsion respectively.The results are shown in Table three.
Table three
| The sampling batch | - | Four | Seven | Ten |
| Protein concn | 3.73％ | 3.87％ | 3.90％ | 3.88％ |
| Solid concentration | 6.37％ | 6.51％ | 6.48％ | 6.50％ |
Presentation of results: the continuous processing extraction effect is stable, can remain good extraction yield.This continuous extraction is industrial feasible.Extractor volume ratio interrupter method is littler, and the abstraction reaction time can shorten to 1.0 hours.
Test example 4, continuous precipitation effect
To test example 3 collections and separate the whole protein emulsions that obtain that remove slag and in time cool off, merge and be stored in room temperature, raw material as test example 4, and measuring wherein with Kjeldahl method, protein concentration is 3.87%, with doing the precipitation experiment with sampling device with experimental example 3, first 1200 milliliters of emulsions are in 50 ℃ of stirring reactions, dropping at 4.5 o'clock to the pH value begins to exchange new emulsion with 20 milliliters of speed of per minute, slurry pH value in the reactor maintains about 4.5 at this moment, every collection is a collection of for 1200 milliliters, centrifugation goes out protein precipitation and whey liquid (being called whey liquid one time), again a whey liquid is heated to about 65 ℃, produce white cotton-shaped albumen,, obtain secondary sedimentation albumen and second whey again through centrifugal.Measure second, four, six, eight batch of protein concentration in whey liquid of totally four batches of samples and the second whey liquid with Kjeldahl determination respectively.Protein precipitation then washs 2 times with the sour water of pH value 4.5, surveys the albumen yield with method.The results are shown in Table four.
Table four
| The sampling batch | Two | Four | Six | Eight |
| The soya-bean milk raw material | 3.87％ | 3.87％ | 3.87％ | 3.87％ |
| A whey-protein concentration | 0.68％ | 0.85％ | 0.85％ | 0.85％ |
| The second whey protein concentration | 0.59％ | 0.59％ | 0.59％ | 0.59％ |
| Primary sedimentation albumen heavy (gram) | 37.6 | 35.2 | 35.4 | 34.9 |
| Secondary sedimentation albumen heavy (gram) | 1.00 | 3.01 | 3.00 | 3.10 |
| Residual soluble protein 0.59% * 1200ml (gram) in the second whey | 7.08 | 7.08 | 7.08 | 7.08 |
From table four measurement result as seen: after entering successive reaction, except that first batch of material, the isolated albumen of four, six, eight batch materials comprises that the albumen that once settles out, the secondary albumen yield that settles out all is stable, illustrates that precipitin reaction is steadily feasible.The numerical value of protein content of settling out have slightly deviation be error at measurment institute extremely, and from once, residual soluble proteins concentration more can illustrate problem the second whey liquid: the hydrolytic precipitation reaction is stable, also is operable approach in the production.
Continuous processing is compared with interrupter method in addition, a certain amount of albumen that solvable heating then can be separated out that do not heat when pH value 4.5 occurred.This part albumen is the low-molecular-weight new constituent of enzyme digestion reaction output.
The soluble protein of this part pH value 4.5 that continuous processing is produced can be through heating centrifugation again, transfers to neutrality, dry or transfer neutrality drying more jointly with protein precipitation separately then.
Test example 5, protein isolate solubility experiment
The protein precipitation that continuous processing precipitation obtains is washed after the centrifugation again with alkaline water dissolving and adjust pH to 7.0-7.5, also heat the enzyme that goes out through homogeneous again, obtain little pale brown look translucent solution, through 3000rpm, the centrifugal precipitation that do not produce of 10min, spray-dried again formation separated protein powder.The protein powder that (centrifugal spray-drying tower: 200 ℃ of inlet temperature, 80-85 ℃ of outlet wind-warm syndrome degree) obtains all can be dissolved into the preceding colloidal sol shape of spraying drying immediately with 80 ℃ of pure water or ordinary tap water, through 3000rpm centrifugal 10 minutes, do not produce precipitation, appearance effect is with spraying drying is preceding not identical, and this protein powder can be used as instant protein drinks batching and uses.Can only obtain suspension liquid and reconstitute with 80 ℃ of pure water by the spray-dired protein powder of similarity condition, place then to produce in a large number and precipitate with the heavy method of acid.
Utilize domestic industry standard to survey molten nitrogen exponential method relatively, the solvable index of separated protein powder nitrogen (NSI) that the present invention utilizes the enzyme precipitator method to obtain is 94%, reaches the requirement (reference to standard is Q/22W004.1-1997) of domestic industry standard 〉=90%.
Above experimental example proves, of the present invention utilize behind the native protein enzyme activation that exists in the soybean as soy proteinaceous chaotropic agent and make the pH value drop to 4.5 through spontaneous enzyme reaction extract the method that prepare soybean protein isolate as precipitation agent and can replace the alkali extraction and acid precipitation that passes through on the our times fully.The characteristics of present method are: 1, it uses the soda acid in the enzyme replacement chemical method; What 2, it used is not zymin, but utilizes the biological enzyme that self exists in the plant seed, makes it to become real biotechnology; 3, according to the principle of the inventor in first to file 02157397.2, the dregs of rice that denaturation degrees is big even the high temperature dregs of rice also can be used as the protein isolate raw material and use; 4, compare with current generic acid alkaline process, reduced the use of chemical feedstocks, reduced pollution, reduced relevant device and raw materials cost that acid, alkali use.Desalination operation and equipment be can save during whey reprocessing, co-production equipment and cost greatly reduced; 5, compare with acid-base method, present method production unit is simplified greatly, and equipment and factory building investment can reduce greatly; 6, the soybean protein isolate of produced in conventional processes solvability in water is very poor, be difficult to form real sol solutions, and present method soybean protein isolate NSI value can reach industry standard both domestic and external; 7, present method is because of the raw meat effect of taking off in the enzymolysis process, so the protein isolate of producing does not have beany flavor.
Embodiment 1
The soybean low temperature soy meal is ground into 120 order fine powders, gets 1000 gram thin bean cake powders and 12 liters of about 50 ℃ of warm water and mixes, and is incubated 50 ℃ and with electric mixer stirring 2 hours, and material liquid pH value drops to 6.28 by 6.48.Feed liquid with small-sized solid bowl centrifuge centrifugation, is obtained 11.5 liters of soya-bean milk (recording wherein, protein concn is 3.67%), 1.25 kilograms in wet bean dregs (water ratio is 82.0%, and protein content is 26.7% in the bean dregs of oven dry back).
The soya-bean milk liquid that is gone out by centrifugation (feeds about 45 ℃ of water insulation) in the interlayer in the double-layer heat insulation bucket, emulsion stirs with electric mixer, and constantly monitor ph value of emulsion and change, reduce to 4.5 through 1 hour 45 minutes pH values, promptly stop to stir, emulsion is with 10.0 liters of whizzer separating whey liquid and 1.42 kilograms of egg white icings (it is 29.7% that solid substance contains rate, and protein is 25.6%).
Sour water with pH value 4.5 washs egg white icing once earlier, again egg white icing is added 4%NaOH solution limit Jia Shui with stirrer while smashing, transferring to pH value 7.0 becomes translucent aqueous, and use the clarifixator homogeneous, can fully disperse the protein particle of dispersing and dissolving not during because of homogeneous, the pH value can descend, again transfer to pH value 7.0 with alkali lye once more, protein concn in the emulsion is 19.2% at this moment, 20 minutes sterilization enzyme inactivations of insulation in 95 ℃ of water-baths, spraying drying (small-sized spray drier: 210 ℃ of air intakes again, go out one's intention as revealed in what one says 80-85 ℃), obtain about 300 grams of soy protein isolate powder (because of the spray-drier loss can't be calculated yield).
This protein powder protein content 88.3%, moisture content 5.4%, ash 3.3%, nitrogen solubility index 91.5% can be dissolved into dissolved colloidal state fully with 60 ℃ of hot water dissolvings, and microscopic be can't see not soluble particles, has only the small quantities of particles fragment.
Change Example
It more than is automatic activation method, as adopt other two kinds of activation methods, this embodiment then changes: according to the described method preparation " activator " earlier of embodiment 1 or embodiment 2 in first to file 02157397.2, add then in the above-mentioned beans paste, abstraction reaction is carried out in stirring, observe the pH value and change to meet the requirements (this extraction time will shorten in about 1 hour from 2 hours), later operation is the same.Precipitin reaction also can shorten in 1 hour.
Embodiment 2
3 kilograms of low temperature soy meals add 33 liters of room temperature tap water to be worn into the beans paste and adds defoamer with colloidal mill, uses 4%NaOH solution adjust pH in 6.8 again, preserves standby in room temperature (18-25 ℃).
In 55 ℃ of water-baths, be incubated and start electric mixer and stir (120rpm) with the stainless steel cask of the be back to back feed liquid of 8 liters of real volumes, when beans paste pH value drops to 6.4, start charging/discharging device, and keep by regulating input and output material speed that material liquid pH value is stable at 6.4 in the bucket.Under this condition per 65 minutes, material is once commutative in the bucket, the material of discharging also stirs (120rpm) in 45 ℃ of water bath heat preservations through the soya-bean milk liquid that centrifugation removal bean dregs obtain in another set of 6 liters continuous reaction apparatus, when reducing to 4.5 by 6.4, soya-bean milk pH value opens charging/discharging device, turnover soya-bean milk speed to be keeping the pH value stabilization in 4.5 degree of being, the pH value of discharge is that 4.5 soya-bean milk carries out centrifugation.The material exchange is once about 50 minutes in this precipitin reaction device.
Enter normal switching phase post-extraction tank discharge velocity and setting tank input speed basically identical.Under production status, the centre should be provided with the buffering basin, to regulate the balance of two-step reaction.
The pH value reaches 4.5 soya-bean milk and obtains egg white icing mud and whey liquid through centrifugation, whey liquid produces white cotton-shaped albumen precipitation through heating (60-65 ℃), obtain the secondary egg white icing with filter cloth, merge with primary sedimentation albumen, through once with the water washing of pH value 4.5, add 4%NaOH solution and water while smashing with stirrer again, be transferred to pH value about 8.0, treat that egg white icing returns to liquid, the pH value descends behind the clarifixator homogeneous, this is that the protein precipitation particle does not disperse fully when smashing because stir, can not get neutralization, material pH value descends again after clarifixator is smashed, and behind the adjust pH to 7.0, is heated to 95 ℃ once more, stirring also keeps 20 minutes sterilization enzyme inactivations, spraying drying (centrifugal spray-drying tower: 230 ℃ of inlet temperature, temperature of outgoing air 80-85 ℃), obtain about 1 kilogram of separated protein powder (the spraying drying loss is bigger, can not calculate accurate yield).This protein isolate protein content 87.4%, moisture 5.7%, ash 3.6%, molten nitrogen index 90.5% can be dissolved into collosol state during with 70 ℃ of hot water dissolvings immediately.
This embodiment changes one: with the Change Example of embodiment 1.
This embodiment changes two: since second batch materials, change into using and mix dregs of beans, that is: the weight ratio of the high temperature dregs of rice and the low temperature dregs of rice is 80: 20-50: 50, and the albumen that obtains is in not obviously difference in nature, and just protein extracting ratio has reduction slightly.
The method for making of embodiment 3, peanut protein isolate
Three kinds of Activiation methods in the proteinase activated method (application number 02157397.2) in the oil crops seed of applying in advance by the inventor, or can use any method and any feedstock production activator such as peanut, the peanut low temperature dregs of rice, soybean, soybean meal earlier.
Adopt the method for embodiment 1 or embodiment 2 to produce separating protein powder of peanut equally.
Activator preparation: get 300 8-12 hour peanuts of gram immersion and 1.5 premium on currency and wear into slurry back insulation and electronic stirring (100rpm) in 55 ℃ of water-baths, the pH value begins to descend after 3-3.5 hour, drip 4%NaOH solution again and keep pH value stabilization continuation reaction 1 hour, standby as activator.
3 kilograms of peanut meal fine powders (containing protein 45.7%) and 24 liters of tap water furnishing pasty states, it is standby that adjust pH 6.8 is stored in room temperature (18-25 ℃).
Same apparatus and method with embodiment 2: in extraction tank, add activator and add peanut meal powder aqueous dispersions, in 50 ℃ of insulations and stirring.When reducing to 6.2, material pH value begins constantly to discharge reaction material and constantly adding virgin material, the centrifugal slagging-off of the material of collecting, the peanut emulsion enters settling bath in 45 ℃ of insulations and electronic stirring (100rpm), when material pH value drops to 4.7, peanut emulsion from settling bath after continuous discharge reaction solution and additional the removing slag is finished tandem reaction sequence.Whole process and embodiment 2 identical (amount of water is 8 times, isoelectric pH value 4.7).
The Semen arachidis hypogaeae protein of centrifugation is through washing, neutralization, homogeneous, adjust pH, sterilization, spraying drying obtain 0.9 kilogram of peanut protein powder (spraying drying loss can't calculated yield) again.Protein content 92%, solvability is better than soybean isolate protein powder.
Claims (8)
1, the manufacture method of plant separation protein, it is characterized in that, at first activate the contained proteolytic ferment of raw material self, make raw material issue unboiled water in the effect of this proteolytic ferment and separate reaction and obtain protein emulsion, when the pH of this emulsion value reached 6.5-6.0, the material slag was removed in separation, make protein emulsion continue to be hydrolyzed into pH value 5.0-3.0, separate and collect protein precipitation and whey liquid, protein precipitation is through transferring neutrality, and drying becomes separated protein powder;
Wherein, the process that activates the contained proteolytic ferment of raw material self comprises one of following method of employing preparation activator, utilizes this activator to start the hydrolysis reaction of raw material then:
1) utilize any protease preparation to make under optimum conditions after the raw material slurry generation hydrolysis as activator;
2) with raw slurry at 30-65 ℃, stirred under the pH value 6.5-8.5 condition 2-6 hour, during pH value keep by adding alkaline agent, the raw material oneself protease is activated afterwards as activator;
3) protein emulsion that obtains after preceding a collection of raw water is separated, the perhaps direct activator of whey liquid behind its continuation hydrolysis precipitation separation albumen as following batch raw material.
2, the described manufacture method of claim 1, wherein, the raw material of used production plant separation protein is soybean, genetically engineered soybean, low temperature soy meal, high temperature dregs of beans, peanut meal or its mixture.
3, the described manufacture method of claim 1, wherein, the raw material of preparation activator is soybean, peanut, sesame, almond, walnut kernel, sunflower seed, Semen Brassicae campestris, cotton seed or their the low temperature dregs of rice.
4, the described manufacture method of claim 1, wherein, raw material under the effect of oneself protein lytic enzyme the hydrolysis abstraction reaction and protein emulsion to continue the temperature control that hydrolysis obtains the precipitin reaction of protein precipitation be the upper limit with the proteolytic enzyme non-inactivation all.
5, the described manufacture method of claim 4, wherein, the temperature 30-65 of hydrolysis abstraction reaction ℃, the temperature and the hydrolysis reaction of precipitin reaction are identical or different.
6, the described manufacture method of claim 1, wherein, enzyme extraction and/or precipitation process adopt continuation method, after the material in hydrolysis extractor and the precipitin reaction jar meets the requirements of the pH scope, constant speed adds new feed liquid when discharging, and input and output material speed is as the criterion to keep a jar interior pH value stabilization.
7, the described manufacture method of claim 1, wherein, when protein emulsion continued hydrolysis generation precipitin reaction, control pH finished when value drops to 5.0-4.0.
8, the described manufacture method of claim 1, wherein, the whey liquid heating behind the precipitation separation albumen makes albumen precipitation wherein once more, collect this secondary sedimentation albumen, be adjusted to neutrality, drying becomes protein powder, perhaps, this secondary sedimentation albumen and protein precipitation merging are last time handled.
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2003101208352A CN1318603C (en) | 2003-12-30 | 2003-12-30 | Method for preparing plant separated protein |
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| Application Number | Priority Date | Filing Date | Title |
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| CNB2003101208352A CN1318603C (en) | 2003-12-30 | 2003-12-30 | Method for preparing plant separated protein |
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| CN1635136A CN1635136A (en) | 2005-07-06 |
| CN1318603C true CN1318603C (en) | 2007-05-30 |
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| CNB2003101208352A Expired - Lifetime CN1318603C (en) | 2003-12-30 | 2003-12-30 | Method for preparing plant separated protein |
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN101558814B (en) * | 2009-05-15 | 2011-10-12 | 山东省高唐蓝山集团总公司 | Production technology of dual-crop compound protein isolates |
| CN103190522B (en) * | 2013-04-12 | 2014-02-12 | 山东方健制药有限公司 | Method for extracting protein from sesame |
| CN103190523A (en) * | 2013-04-27 | 2013-07-10 | 西北大学 | Yangtao seed meal isolated protein and extraction method thereof |
| CN103766574A (en) * | 2013-12-31 | 2014-05-07 | 山东省花生研究所 | Process for producing peanut protein isolate by virtue of enzymatic modification |
| WO2017177490A1 (en) * | 2016-04-11 | 2017-10-19 | 浙江省农业科学院 | Method for preparing bean sprout-derived functional peptide |
| CN107927319B (en) * | 2017-12-26 | 2021-06-22 | 南京爱美倍健生物科技有限公司 | Method for preparing peanut polypeptide liquid by hydrolyzing peanut protein by using peanut inherent protease |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0408063A1 (en) * | 1989-07-14 | 1991-01-16 | Cpc International Inc. | A process for the production of hydrolyzed vegetable proteins and the product therefrom |
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Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| EP0408063A1 (en) * | 1989-07-14 | 1991-01-16 | Cpc International Inc. | A process for the production of hydrolyzed vegetable proteins and the product therefrom |
Non-Patent Citations (1)
| Title |
|---|
| 水解植物蛋白(HVP)的酶法制备及应用 武彦文等，中国调味品，第279期 2002 * |
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