CN103988975A - Method for extracting protein from soybean meal by adopting ultrasonic-assisted reverse micelle extraction process - Google Patents
Method for extracting protein from soybean meal by adopting ultrasonic-assisted reverse micelle extraction process Download PDFInfo
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- CN103988975A CN103988975A CN201410232504.6A CN201410232504A CN103988975A CN 103988975 A CN103988975 A CN 103988975A CN 201410232504 A CN201410232504 A CN 201410232504A CN 103988975 A CN103988975 A CN 103988975A
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Abstract
The invention discloses a method for extracting protein from soybean meal by adopting an ultrasonic-assisted reverse micelle extraction process, and belongs to plant protein extraction and processing technologies. The method comprises the following steps: (1) grinding defatted soybean meal, screening, mixing with water to form a mixed solution, adding cellulase and protease into the mixed solution in sequence, performing step-by-step enzymatic hydrolysis, and after the enzymatic hydrolysis, performing centrifugal separation to obtain enzymatic hydrolysis liquid and deposits; (2) adding a reverse micelle system into the enzymatic hydrolysis liquid for performing ultrasonic-assisted reverse micelle extraction to obtain extracting liquid, performing back extraction on the extracting liquid, and performing the centrifugal separation to obtain an organic phase and a water phase; (3) performing alkali-solution and acid-isolation on the water phase to obtain deposits, adding the water into the deposits for redissolving, ultra-filtering, concentrating in vacuum and freeze-drying to obtain the soybean protein. According to the method, required process equipment is simple and safe to operate; an ultrasonic technology and a reverse micelle extraction technology are combined for extracting the protein from the soybean meal, so that the protein extraction rate is increased and the extraction cost is reduced; the prepared soybean protein is high in purity, low in modification degree and good in functionality.
Description
Technical field
The invention belongs to the extraction process technology of vegetable protein, relate generally to the method for albumen in a kind of ultrasonic auxiliary inverse micelle abstraction dregs of beans.
Background technology
In dregs of beans, generally contain the multiple nutritional components such as albumen, polysaccharide, fiber, especially contain abundant albumen, content is very high, and this is also the composition that value is the highest.In dregs of beans albumen, containing the multiple necessary amino acid of a large amount of needed by human body, is also therefore very important food protein source.But most of dregs of beans are all used as feed, this has just caused the significant wastage of soybean protein resource.
The research of inverse micelle abstraction albumen, since 20 century 70s, develops into and has had now 30 years.The mechanism that it has been generally acknowledged that inverse micelle abstraction protein is that water-solubility protein is dissolved in the pond of reverse micelle system.Reverse micelle system is in dynamic equilibrium, interior internuclearly ceaselessly bump, and often exchange the contained material of kernel.While utilizing reverse micelle to extract protein, protein depends primarily on the condition of water in two alternate distribution, as pH value, ionic strength, electrolyte type; Organic phase condition also affects protein in two alternate distribution, as kinds of surfactants, and type of organic solvent, cosurfactant; The character of temperature and protein self etc. also has influence on the transfer of protein at inverse micellar solution.Mainly extract protein with liquid-liquid extraction at present, owing to there being micro-pond in reverse micelle, therefore biologically active such as solubilized amino acid, point peptides and proteins can not make its inactivation, but the extraction yield of reverse micelle are not desirable especially.
Ultrasonic wave is a kind of elastic mechanical ripple in material medium, and it produces fuel factor, mechanical effect or cavitation effect in the time of Propagation.Research thinks, the reason that ultrasonic wave can improve aqueous chemical reaction speed and productive rate is mainly the cavitation of ultrasonic wave in liquid medium.Cavitation can cause the second order effects such as turbulence effect, perturbation effect, interfacial effect, Mohaupt effect.Due to the synergy of various effects, make ultrasonic wave can strengthen the motive force of inverse micelle abstraction soybean protein process, and then improve the extraction yield of soybean protein.
Ultrasonic technology and inverse micelle abstraction technique are combined use by the inventive method, be applied to the extraction of dregs of beans albumen, under hyperacoustic cavitation effect effect, strengthen the motive force of inverse micelle abstraction soybean protein process, improve protein extracting ratio, reduced extraction cost, also ensured purity and the activity of albumen simultaneously, for the actual production of extracting soybean protein in dregs of beans is provided fundamental basis, this recycling and minimizing wasting of resources to dregs of beans has great importance.
Summary of the invention
Technical problem to be solved by this invention is to overcome the deficiencies in the prior art, and the method for albumen in a kind of ultrasonic auxiliary inverse micelle abstraction dregs of beans is provided, and reaches the object that improves protein extracting ratio, purity and protected protein activity.
Technical problem to be solved by this invention is achieved through the following technical solutions:
A kind of method of albumen in ultrasonic auxiliary inverse micelle abstraction dregs of beans, the method comprises the following steps: (1) defatted soybean meal is pulverized and sieved afterwards and water is mixed to form mixed liquor, first in mixed liquor, add cellulase to carry out enzymolysis, described cellulase addition is 2% of mixed liquor quality, hydrolysis temperature is 50 DEG C, enzymolysis time is 1.5h, enzymolysis pH is 5.5, in mixed liquor, add alkali protease to carry out enzymolysis again, described alkali protease addition is 3% of mixed liquor quality, hydrolysis temperature is 60 DEG C, enzymolysis time is 2h, enzymolysis pH is 9, after enzymolysis, centrifugation obtains enzymolysis liquid and precipitation, (2) in enzymolysis liquid, add AOT/ isooctane reverse micelle system to carry out ultrasonic auxiliary inverse micelle abstraction and be extracted liquid, described surfactant AOT concentration is 0.02-0.1g/mL, ionic strength is 0.05-0.25mol/L KCl, ultrasonic power is 150-350W, extraction temperature is 30-70 DEG C, extraction time is 0.5-2.5h, extraction pH is 6-10, in extract, add the KCl solution of isopyknic 1mol/L to strip, described reextraction temperature is 30-50 DEG C, back-extraction time is 10-50min, reextraction pH is 6-10, after stripping, centrifugation obtains organic phase and water, (3) water pH is adjusted to alkalescence and carries out alkali and carry, then regulate pH to carry out acid to acidity heavy, the precipitation obtaining is added water after redissolving and carries out hyperfiltration treatment, ultrafiltration final vacuum is concentrated, freeze drying obtains soybean protein.
Described ultrasonic auxiliary inverse micelle abstraction preferred parameter is: surfactant AOT concentration 0.06g/mL, ionic strength 0.1mol/L KCl, ultrasonic power 250W, 50 DEG C of extraction temperature, extraction time 1h, extraction pH7.
Described reextraction preferred parameter is: 40 DEG C of reextraction temperature, back-extraction time 30min, reextraction pH9.
This method is on to the basis of defatted soybean meal substep enzymolysis, adopt ultrasonic auxiliary inverse micelle abstraction dregs of beans albumen, activity that can Protein requirement in reverse micelle system, combining ultrasonic aid in treatment again, strengthen the motive force of inverse micelle abstraction soybean protein process, make protein be easier to enter into reverse micelle system pond, extraction yield is increased greatly.The method has that required process equipment is simple, the feature of handling safety, and extraction rate of protein is high, and the soybean protein purity of acquisition is high, denaturation degrees is low, functional good.
Brief description of the drawings
Accompanying drawing overall process route figure of the present invention.
Detailed description of the invention
Below in conjunction with accompanying drawing, the specific embodiment of the invention is described in detail,
A kind of method of albumen in ultrasonic auxiliary inverse micelle abstraction dregs of beans, the method comprises the following steps: (1) defatted soybean meal is pulverized and sieved afterwards and water is mixed to form mixed liquor, first in mixed liquor, add cellulase to carry out enzymolysis, described cellulase addition is 2% of mixed liquor quality, hydrolysis temperature is 50 DEG C, enzymolysis time is 1.5h, enzymolysis pH is 5.5, in mixed liquor, add alkali protease to carry out enzymolysis again, described alkali protease addition is 3% of mixed liquor quality, hydrolysis temperature is 60 DEG C, enzymolysis time is 2h, enzymolysis pH is 9, after enzymolysis, centrifugation obtains enzymolysis liquid and precipitation, (2) in enzymolysis liquid, add AOT/ isooctane reverse micelle system to carry out ultrasonic auxiliary inverse micelle abstraction and be extracted liquid, described surfactant AOT concentration is 0.02-0.1g/mL, ionic strength is 0.05-0.25mol/L KCl, ultrasonic power is 150-350W, extraction temperature is 30-70 DEG C, extraction time is 0.5-2.5h, extraction pH is 6-10, in extract, add the KCl solution of isopyknic 1mol/L to strip, described reextraction temperature is 30-50 DEG C, back-extraction time is 10-50min, reextraction pH is 6-10, after stripping, centrifugation obtains organic phase and water, (3) water pH is adjusted to alkalescence and carries out alkali and carry, then regulate pH to carry out acid to acidity heavy, the precipitation obtaining is added water after redissolving and carries out hyperfiltration treatment, ultrafiltration final vacuum is concentrated, freeze drying obtains soybean protein.
Described ultrasonic auxiliary inverse micelle abstraction preferred parameter is: surfactant AOT concentration 0.06g/mL, ionic strength 0.1mol/L KCl, ultrasonic power 250W, 50 DEG C of extraction temperature, extraction time 1h, extraction pH7.
Described reextraction preferred parameter is: 40 DEG C of reextraction temperature, back-extraction time 30min, reextraction pH9.
Embodiment 1:
By defatted soybean meal pulverize and sieve after and water be mixed to form mixed liquor, it is 50 DEG C at hydrolysis temperature, enzymolysis pH carries out enzymolysis 1.5h to the cellulase that adds 2% in mixed liquor 5.5 times, then it is 60 DEG C at hydrolysis temperature, enzymolysis pH carries out enzymolysis 2h to the alkali protease that adds 3% in mixed liquor 9 times, after enzymolysis, centrifugation obtains enzymolysis liquid and precipitation, be under 0.1mol/L KCl in ionic strength, the surfactant AOT that is 0.06g/mL by concentration and isooctane are mixed to form AOT/ isooctane reverse micelle system, in enzymolysis liquid, add AOT/ isooctane reverse micelle system, be 250W at ultrasonic power, extraction temperature is 50 DEG C, extraction pH carries out ultrasonic auxiliary inverse micelle abstraction 1h for 7 times to be extracted liquid, it is 40 DEG C in reextraction temperature, reextraction pH is 9 times, to the KCl solution that adds isopyknic 1mol/L in the extract 30min that strips, after stripping, centrifugation obtains organic phase and water, water pH is adjusted to alkalescence to carry out alkali and carries, then regulate pH to carry out acid to acidity heavy, by the precipitation obtaining add water redissolve after carry out hyperfiltration treatment, ultrafiltration final vacuum is concentrated, freeze drying obtains soybean protein.Soybean protein recovery rate prepared by the method is 91.58%, and the soybean protein purity of gained is high, denaturation degrees is low, functional good, and has certain biologically active.
Embodiment 2:
By defatted soybean meal pulverize and sieve after and water be mixed to form mixed liquor, it is 50 DEG C at hydrolysis temperature, enzymolysis pH carries out enzymolysis 1.5h to the cellulase that adds 2% in mixed liquor 5.5 times, then it is 60 DEG C at hydrolysis temperature, enzymolysis pH carries out enzymolysis 2h to the alkali protease that adds 3% in mixed liquor 9 times, after enzymolysis, centrifugation obtains enzymolysis liquid and precipitation, be under 0.05mol/L KCl in ionic strength, the surfactant AOT that is 0.08g/mL by concentration and isooctane are mixed to form AOT/ isooctane reverse micelle system, in enzymolysis liquid, add AOT/ isooctane reverse micelle system, be 300W at ultrasonic power, extraction temperature is 60 DEG C, extraction pH carries out ultrasonic auxiliary inverse micelle abstraction 1.5h for 8 times to be extracted liquid, it is 45 DEG C in reextraction temperature, reextraction pH is 8 times, to the KCl solution that adds isopyknic 1mol/L in the extract 40min that strips, after stripping, centrifugation obtains organic phase and water, water pH is adjusted to alkalescence to carry out alkali and carries, then regulate pH to carry out acid to acidity heavy, by the precipitation obtaining add water redissolve after carry out hyperfiltration treatment, ultrafiltration final vacuum is concentrated, freeze drying obtains soybean protein.Soybean protein recovery rate prepared by the method is 90.69%, and the soybean protein purity of gained is high, denaturation degrees is low, functional good, and has certain biologically active.
Embodiment 3:
By defatted soybean meal pulverize and sieve after and water be mixed to form mixed liquor, it is 50 DEG C at hydrolysis temperature, enzymolysis pH carries out enzymolysis 1.5h to the cellulase that adds 2% in mixed liquor 5.5 times, then it is 60 DEG C at hydrolysis temperature, enzymolysis pH carries out enzymolysis 2h to the alkali protease that adds 3% in mixed liquor 9 times, after enzymolysis, centrifugation obtains enzymolysis liquid and precipitation, be under 0.1mol/L KCl in ionic strength, the surfactant AOT that is 0.06g/mL by concentration and isooctane are mixed to form AOT/ isooctane reverse micelle system, in enzymolysis liquid, add AOT/ isooctane reverse micelle system, be 300W at ultrasonic power, extraction temperature is 55 DEG C, extraction pH carries out ultrasonic auxiliary inverse micelle abstraction 1h for 9 times to be extracted liquid, it is 35 DEG C in reextraction temperature, reextraction pH is 8 times, to the KCl solution that adds isopyknic 1mol/L in the extract 35min that strips, after stripping, centrifugation obtains organic phase and water, water pH is adjusted to alkalescence to carry out alkali and carries, then regulate pH to carry out acid to acidity heavy, by the precipitation obtaining add water redissolve after carry out hyperfiltration treatment, ultrafiltration final vacuum is concentrated, freeze drying obtains soybean protein.Soybean protein recovery rate prepared by the method is 89.88%, and the soybean protein purity of gained is high, denaturation degrees is low, functional good, and has certain biologically active.
Claims (3)
1. the method for albumen in a ultrasonic auxiliary inverse micelle abstraction dregs of beans, it is characterized in that the method comprises the following steps: (1) defatted soybean meal is pulverized and sieved afterwards and water is mixed to form mixed liquor, first in mixed liquor, add cellulase to carry out enzymolysis, described cellulase addition is 2% of mixed liquor quality, hydrolysis temperature is 50 DEG C, enzymolysis time is 1.5h, enzymolysis pH is 5.5, in mixed liquor, add alkali protease to carry out enzymolysis again, described alkali protease addition is 3% of mixed liquor quality, hydrolysis temperature is 60 DEG C, enzymolysis time is 2h, enzymolysis pH is 9, after enzymolysis, centrifugation obtains enzymolysis liquid and precipitation, (2) in enzymolysis liquid, add AOT/ isooctane reverse micelle system to carry out ultrasonic auxiliary inverse micelle abstraction and be extracted liquid, described surfactant AOT concentration is 0.02-0.1g/mL, ionic strength is 0.05-0.25mol/L KCl, ultrasonic power is 150-350W, extraction temperature is 30-70 DEG C, extraction time is 0.5-2.5h, extraction pH is 6-10, in extract, add the KCl solution of isopyknic 1mol/L to strip, described reextraction temperature is 30-50 DEG C, back-extraction time is 10-50min, reextraction pH is 6-10, after stripping, centrifugation obtains organic phase and water, (3) water pH is adjusted to alkalescence and carries out alkali and carry, then regulate pH to carry out acid to acidity heavy, the precipitation obtaining is added water after redissolving and carries out hyperfiltration treatment, ultrafiltration final vacuum is concentrated, freeze drying obtains soybean protein.
2. the method for albumen in a kind of ultrasonic auxiliary inverse micelle abstraction dregs of beans according to claim 1, it is characterized in that described ultrasonic auxiliary inverse micelle abstraction preferred parameter is: surfactant AOT concentration 0.06g/mL, ionic strength 0.1mol/L KCl, ultrasonic power 250W, 50 DEG C of extraction temperature, extraction time 1h, extraction pH7.
3. the method for albumen in a kind of ultrasonic auxiliary inverse micelle abstraction dregs of beans according to claim 1, is characterized in that described reextraction preferred parameter is: 40 DEG C of reextraction temperature, back-extraction time 30min, reextraction pH9.
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Cited By (4)
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| CN104509679A (en) * | 2014-11-24 | 2015-04-15 | 威海市桢昊生物技术有限公司 | Method for preparation of soybean glycoprotein from defatted soybean meal |
| CN108586603A (en) * | 2018-04-23 | 2018-09-28 | 西安德氏禾本生物科技有限公司 | A method of preparing the Elastin peptide containing a variety of peculiar amino acid |
| CN109170122A (en) * | 2018-07-27 | 2019-01-11 | 咸阳师范学院 | A kind of extracting soybean albumen simultaneously purifies and the method for recycling surfactant |
| CN113388511A (en) * | 2021-07-02 | 2021-09-14 | 泉州市嗣家名茶发展有限公司 | Device and method for extracting plant protein from okra fruits by bundle extraction method |
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| CN101343309A (en) * | 2008-08-25 | 2009-01-14 | 河南工业大学 | Method for simultaneously separating soy protein and oil fat with inverse micelle abstraction technique |
| CN101912030A (en) * | 2010-09-26 | 2010-12-15 | 河南工业大学 | Method for extracting soybean protein by using dodecyltrimethylammonium chloride reverse micellar solution |
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104509679A (en) * | 2014-11-24 | 2015-04-15 | 威海市桢昊生物技术有限公司 | Method for preparation of soybean glycoprotein from defatted soybean meal |
| CN104509679B (en) * | 2014-11-24 | 2017-08-18 | 威海市桢昊生物技术有限公司 | A kind of method that utilization defatted soybean meal produces soy glycoproteins |
| CN108586603A (en) * | 2018-04-23 | 2018-09-28 | 西安德氏禾本生物科技有限公司 | A method of preparing the Elastin peptide containing a variety of peculiar amino acid |
| CN109170122A (en) * | 2018-07-27 | 2019-01-11 | 咸阳师范学院 | A kind of extracting soybean albumen simultaneously purifies and the method for recycling surfactant |
| CN113388511A (en) * | 2021-07-02 | 2021-09-14 | 泉州市嗣家名茶发展有限公司 | Device and method for extracting plant protein from okra fruits by bundle extraction method |
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