CN103146788A - Enzymic-method based preparation method of soybean peptide - Google Patents
Enzymic-method based preparation method of soybean peptide Download PDFInfo
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- CN103146788A CN103146788A CN2013100645204A CN201310064520A CN103146788A CN 103146788 A CN103146788 A CN 103146788A CN 2013100645204 A CN2013100645204 A CN 2013100645204A CN 201310064520 A CN201310064520 A CN 201310064520A CN 103146788 A CN103146788 A CN 103146788A
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Abstract
The invention provides an enzymic-method based preparation method of soybean peptide, wherein earthworm serine protease is creatively employed as zymogen for enzymolysis of soybean, thereby separating protein, wheat protein and the like; the hydrolysis efficiency is high; the ultrafiltration technology is employed in the preparation process so that the obtained enzymolysis product soybean peptide is separated more thoroughly; the yield is high; the conditions of the whole process are moderate; and the method is environment-friendly.
Description
Technical field
The present invention relates to the preparation technology of soybean peptides, refer to especially a kind of enzymatic-process preparation method of soybean peptides.
Background technology
Utilize soybean protein after enzymic hydrolysis, obtain the mixed hydrolysis product that formed by multiple peptide molecule, be called soybean peptides.Soybean peptides and soybean protein have identical amino acid and form, and generally are made of 2～10 amino-acid residues, and balance of essential amino acids is good.Compare with soybean protein, soybean peptides possesses good processing characteristics and unique physiologically active, can be used in the production of numerous food and even makeup.
The reaction that is hydrolyzed at a certain temperature of the research original adoption soda acid chemical reagent of soybean protein hydrolyate, but find under study for action, chemical method prepares soybean peptides and has all drawbacks, as wayward in hydrolytic process, hydrolysis is many without specificity, side reaction, nutritive ingredient loss and the problems such as the hydrolysate organoleptic properties is poor, bitter stink are failed solution etc., the progress of soybean oligopeptide is slow.
Along with the zymetology industrial expansion, the enzymatic hydrolysis soybean protein becomes the main flow of research gradually.The processing condition that enzyme process prepares soybean polypeptide are generally all gentleer, but the difference because of the enzyme physicochemical property, its reaction conditions must be limited in certain optimum temperuture and pH scope, otherwise enzyme will inactivation, and action site and the mechanism of action during different enzymic hydrolysis are also different.Alcalase as domestic and international most study is a kind of non-specific alkaline proteolytic enzyme, belongs to restriction endonuclease, and Main Function is in the peptide bond that contains the hydrophobicity carboxyl, and catalytic site is Serine; The active centre of compound protease Protamex is serine residue, has simultaneously the characteristic of restriction endonuclease and excision enzyme, uses more in the soybean peptides enzymolysis; Research is found, the Flavourzyme compound protease can reduce the caused bitter taste of hydrophobic amino acid, can make soybean peptides obtain to use more widely in foodstuffs industry is produced, in recent years, domestic also development to some extent of relevant production technique to soybean peptides, general preparation method is for utilizing the full fishy smell-eliminating soybean protein powder as raw material, use a kind of in Asl.398 proteolytic enzyme, trypsinase, bromeline, process under 30-50 ℃, different pH, get supernatant liquor and be combined into amino acids oral-liquor with amino acid, protein etc. again; Adopt the method for the two enzyme composite hydrolysis of neutral protease Asl.398 and papoid, can obtain to obtain molecular weight in the soybean peptides below 2000 than being used alone the higher degree of hydrolysis of enzyme; Adopt trypsinase and neutral protease Asl.398 composite hydrolysis soybean protein isolate, also obtained good effect, degree of hydrolysis can reach more than 30%, and the soybean peptides mean chain length is 2-4 amino acid, and relative molecular weight is less than 1000, but, the degree of hydrolysis of the conventional single enzymatic hydrolysis of employing is usually lower, is 40-50%, although multi-enzymatic hydrolysis can improve, but still limited, and multi-enzymatic hydrolysis is stricter for the control requirement of processing condition.
The earthworm serine protease is to extract a kind of aspartic protease that obtains in the earthworm body, belongs to trypsinase, its energy protein hydrolysate substrate (as casein), the ester of basic aminoacids and acid amides.PI3-4; 20-50 ℃ of scope is stable; It is a kind of glycoprotein enzyme.
The earthworm serine protease is as a histone lytic enzyme, and is deep not enough to the research of its zymologic property.Most earthworm serine protease components are subjected to the inhibition of 3 kinds of serpin phenylmethylsulfonyl fluorides, tolylsulfonyl phenylalanyl chloromethyl ketone, N-tolysulfonyl lysyl chloromethyl ketone, but different components there are differences the susceptibility of different types of proteinase inhibitor; The earthworm serine protease directly cut Profibrinolysin and fibrin degradation former be its molecular basis that has simultaneously kinases and plasmin activity; The synthetic substrate of difference is had specificity illustrate that it has the different activities center; The earthworm serine protease can have hydrolytic action to the specificity substrate S-2288 of tissue plasminogen activator (t-P A), illustrates that its active mode to Profibrinolysin is similar to t-P A.
Summary of the invention
The present invention proposes a kind of enzymatic-process preparation method of soybean peptides, has improved greatly the efficient of hydrolysis, and the product separation that obtains simultaneously is more thorough, and yield is high.
Technical scheme of the present invention is achieved in that a kind of enzymatic-process preparation method of soybean peptides, comprises the following steps:
(1) take vegetable-protein, according to material ratio 1:20 to 1:60(w/v) add purified water, 75-85 ℃ of sterilization 20-40min is cooled to 40 ℃;
(2) add the earthworm serine protease by vegetable-protein weight 0.5-2 ‰;
(3) enzymolysis process condition: make PH be controlled in the scope of 6.0-7.8 by damping fluid, 37-42 ℃ of insulation hydrolysis 20-30h, then 75-85 ℃ of deactivation 30min filter, and got cleaner liquid;
(4) using molecular weight cut-off is that 10000 ultra-filtration membrane carries out ultrafiltration, adds 2-3 purified water continuation ultrafiltration;
(5) the ultrafiltration peritoneal effluent is carried out drying, get soy peptide powder.
Preferably, in step (1), described material ratio is 1:40, and sterilising temp is 80 ℃, 30min.If the too high sex change that easily causes protein of temperature.
Preferably, in step (2), the weight that adds the earthworm serine protease is 1 ‰ of vegetable-protein weight.
Preferably, in step (3), PH is controlled at 6.5, and the temperature of insulation hydrolysis is 40 ℃, hydrolysis 24h, and the deactivation temperature is 80 ℃.
Preferably, in step (3), described filtration step comprises: the multilayer filtered through gauze, got cleaner liquid; Filter cake adds the purified water washing again, and the multilayer filtered through gauze is crossed cleaner liquid with twice and merged;
Preferably, in step (4), described ultra-filtration membrane is hollow-fibre membrane or rolled film or tubular membrane or ceramic membrane, and certainly, available ultrafiltration apparatus is not limited to this, as long as can reach the ultrafiltration apparatus of effect same, all can use.
Preferably, in step (5), described dry lyophilize or the spraying drying of adopting adopts lyophilize or spraying drying can prevent protein generation sex change.
Preferably, in step (1), described vegetable-protein is soybean protein isolate or wheat protein, but is not limited to this, selects the main difference of different vegetable-proteins to be the difference of efficiency of pcr product, and the title of peptide changes with botanical name simultaneously.
Other common process are controlled and are adopted prior art.
Beneficial effect of the present invention is: the invention utilize the earthworm serine protease as the enzymatic hydrolysis vegetable-protein, especially the proenzyme of soybean protein isolate, its hydrolysis efficiency significantly improves, reach more than 80%, apparently higher than other known enzymes (being generally 40-50%), the product peptide content is high, reaches more than 80%; Also use ultra-filtration technique by adding in the preparation link in the present invention, enzymolysis product is separated thoroughly, yield is high, and the soybean peptides molecular weight that makes is little, biological activity is high, is easier to absorption of human body; The whole technical conditions of the enzymatic-process preparation method of a kind of soybean peptides of the present invention is gentle, environmentally friendly.
Embodiment
For understanding better the present invention; below by following examples, the present invention is done further concrete elaboration; but unintelligible is limitation of the invention; some nonessential improvement and adjustment for those skilled in the art does according to the foregoing invention content also are considered as dropping in protection scope of the present invention.
We adopt method disclosed by the invention, carry out following operation:
(1) take soybean protein isolate 500g, pick up from only rich lucky (Beijing) Science and Technology Ltd., food grade is according to material ratio 1:40(w/v) add purified water 2000ml, 80 ℃ of sterilization 30min(temperature can not be too high, prevents protein denaturation) be cooled to 40 ℃;
(2) add earthworm serine protease 0.5g(source by vegetable-protein weight 1 ‰: Zhuhai Bokang Pharmaceutical Industry Co., Ltd.);
(3) PH is controlled at 6.5,40 ℃ of insulation hydrolysis 24h, 80 ℃ of deactivation 30min; The multilayer filtered through gauze was got cleaner liquid; Filter cake adds the washing of 500ml purified water again, and the multilayer filtered through gauze is crossed cleaner liquid with twice and merged to get 1400ml solution;
(4) using molecular weight cut-off is that 10000 hollow fiber ultrafiltration membrane is carried out ultrafiltration, adds 3 purified water and continues ultrafiltration, gets 2000ml filtrate;
(5) the ultrafiltration peritoneal effluent is carried out lyophilize, get soy peptide powder 17.28g, weight recovery is 35.56%.。
We detect the soybean peptides part that obtains, and wherein, peptide content is 86.0%, higher than the peptide content after the hydrolysis of known other enzymes.
The above is only preferred embodiment of the present invention, and is in order to limit the present invention, within the spirit and principles in the present invention not all, any modification of doing, is equal to replacement, improvement etc., within all should being included in protection scope of the present invention.
Claims (8)
1. the enzymatic-process preparation method of a soybean peptides, is characterized in that, comprises the following steps:
(1) take vegetable-protein, according to material ratio 1:20 to 1:60(w/v) add purified water, 75-85 ℃ of sterilization 20-40min is cooled to 40 ℃;
(2) add the earthworm serine protease by vegetable-protein weight 0.5-2 ‰;
(3) enzymolysis process condition: PH is controlled at 6.0-7.8,37-42 ℃ of insulation hydrolysis 20-30h, and then 75-85 ℃ of deactivation 30min filter, and got cleaner liquid;
(4) using molecular weight cut-off is that 10000 ultra-filtration membrane carries out ultrafiltration, adds 2-3 purified water continuation ultrafiltration;
(5) the ultrafiltration peritoneal effluent is carried out drying, get soy peptide powder.
2. the enzymatic-process preparation method of soybean peptides as described in claim 1, it is characterized in that: in step (1), described material ratio is 1:40, and sterilising temp is 80 ℃, continues 30min.
3. the enzymatic-process preparation method of soybean peptides as described in claim 1, it is characterized in that: in step (2), the weight that adds the earthworm serine protease is 1 ‰ of soybean protein isolate weight.
4. the enzymatic-process preparation method of soybean peptides as described in claim 1, it is characterized in that: in step (3), PH is controlled at 6.5, and the temperature of insulation hydrolysis is 40 ℃ and is hydrolyzed 24h, and the deactivation temperature is 80 ℃.
5. the enzymatic-process preparation method of soybean peptides as described in claim 1, it is characterized in that: in step (3), described filtration step comprises: the multilayer filtered through gauze, got cleaner liquid; Filter cake adds the purified water washing again, and the multilayer filtered through gauze is crossed cleaner liquid with twice and merged.
6. the enzymatic-process preparation method of soybean peptides as described in claim 1, it is characterized in that: in step (4), described ultra-filtration membrane is hollow-fibre membrane or rolled film or tubular membrane or ceramic membrane.
7. the enzymatic-process preparation method of soybean peptides as described in claim 1 is characterized in that: in step (5), and described dry lyophilize or the spraying drying of adopting.
8. the enzymatic-process preparation method of soybean peptides as described in claim 1, it is characterized in that: in step (1), described vegetable-protein is soybean protein isolate or wheat protein.
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Cited By (5)
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| CN103695513A (en) * | 2013-10-14 | 2014-04-02 | 诺利如一（安阳）生物科技有限公司 | Method for improving yield of soybean peptide with low molecular weight |
| CN105941823A (en) * | 2016-05-25 | 2016-09-21 | 刘新旗 | High-soluble soybean peptide dry powder and preparation method thereof |
| CN110122656A (en) * | 2019-06-06 | 2019-08-16 | 南通光合生物技术有限公司 | Soybean protein enzyme hydrolysis process |
| CN110810852A (en) * | 2019-11-04 | 2020-02-21 | 江西沐恩堂生物科技有限公司 | Preparation method of earthworm freeze-dried powder for regulating cardiovascular function |
| CN115466143A (en) * | 2022-08-29 | 2022-12-13 | 河南师范大学 | Earthworm-based amino acid foliar fertilizer and preparation method thereof |
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| CN103695513A (en) * | 2013-10-14 | 2014-04-02 | 诺利如一（安阳）生物科技有限公司 | Method for improving yield of soybean peptide with low molecular weight |
| CN103695513B (en) * | 2013-10-14 | 2016-04-06 | 诺利如一（安阳）生物科技有限公司 | A kind of method improving yield of soybean peptide with low molecular weight |
| CN105941823A (en) * | 2016-05-25 | 2016-09-21 | 刘新旗 | High-soluble soybean peptide dry powder and preparation method thereof |
| CN110122656A (en) * | 2019-06-06 | 2019-08-16 | 南通光合生物技术有限公司 | Soybean protein enzyme hydrolysis process |
| CN110810852A (en) * | 2019-11-04 | 2020-02-21 | 江西沐恩堂生物科技有限公司 | Preparation method of earthworm freeze-dried powder for regulating cardiovascular function |
| CN115466143A (en) * | 2022-08-29 | 2022-12-13 | 河南师范大学 | Earthworm-based amino acid foliar fertilizer and preparation method thereof |
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