CN104212859A - Production method for preparing hydrolyzed wheat proteins by fermentation method - Google Patents

Production method for preparing hydrolyzed wheat proteins by fermentation method Download PDF

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Publication number
CN104212859A
CN104212859A CN201410533095.3A CN201410533095A CN104212859A CN 104212859 A CN104212859 A CN 104212859A CN 201410533095 A CN201410533095 A CN 201410533095A CN 104212859 A CN104212859 A CN 104212859A
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China
Prior art keywords
fermentation
production
hydrolyzed wheat
aspergillus oryzae
hydrolyzed
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CN201410533095.3A
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Inventor
张新武
张子峰
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ZHENGZHOU XINWEI NUTRIENT TECHNOLOGY Co Ltd
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ZHENGZHOU XINWEI NUTRIENT TECHNOLOGY Co Ltd
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Publication of CN104212859A publication Critical patent/CN104212859A/en
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Abstract

The invention discloses a production method for preparing hydrolyzed wheat proteins by a fermentation method, and belongs to the technical field of feed additive production. According to the method, wheat gluten protein powder serves as a raw material and is hydrolyzed into a short peptide chain under the action of protein enzyme preparations such as endo protease and carboxy-peptidase generated by mixed fermentation of bacillus subtilis and aspergillus oryzae, and hydrophobic amino acid at the tail end of the short peptide chain is cut off, so that proteolysis and debittering of the hydrolyzed wheat proteins can be realized at one step by the fermentation method. Compared with a traditional proteolysis method, the production method has the advantages that production cost is low, reaction conditions are moderate, technical parameters are reliable, product quality is stable, bitter taste is avoided, indicators such as product flavor and colors are superior to enzymatic hydrolysis, the hydrolyzed wheat proteins are small in molecular weight, fine in solubility and high in glutamine content, plasma proteins can be replaced, the digestibility of weaning young animals is obviously improved after applications are added, and stress and diarrhea are obviously avoided.

Description

A kind of fermentation method is prepared the production method of hydrolyzed wheat protein
Technical field
The comprehensive exploitation Application Areas that the present invention relates to vegetable-protein, particularly a kind of fermentation method is prepared the production method of hydrolyzed wheat protein, belongs to technical field of feed additive production.
Background technology
The protein molecular weight of wheat-gluten is larger, contain more hydrophobic amino acid and uncharged amino acid, in molecule, hydrophobic interaction region is larger, solubleness is low, after meeting water, easily form gluten shape material, viscosity is large, undesirable or range of application is subject to certain limitation at the effect of all conglomeraties.Hydrolyzed wheat protein as a kind of high-quality without antigen vegetable protein source, be widely used in suckling piglet material, high-grade aquatic products material, calf in milk powder and feed for pet.The production method of hydrolyzed wheat protein mainly contains chemical hydrolysis, enzyme hydrolysis method and microbe fermentation method.Acid and alkali hydrolysis method unstable product quality, the amino acid that hydrolysis produces easily destroys, and easily causes nutritive ingredient loss; Enzyme hydrolysis method is the main method that current hydrolyzed wheat protein is produced, and its weak point is that price is relatively high for the enzyme of gluten powder hydrolysis, and enzymolysis product all exists bitter taste problem to a great extent simultaneously, affects the taste flavor of product and applies; Microbe fermentation method is that the enzyme producing by microbial metabolism activity decomposes gluten powder, decomposes comparatively thoroughly, is rich in the high small-molecular peptides of nutrition.Microbe fermentation method not only cost is low, can save the zymin of a large amount of costlinesses, and product special flavour is good.The domestic and international preparation research to hydrolyzed wheat protein, mainly concentrate at present and utilize pure zymin to carry out direct enzymolysis, because commercial enzyme preparation is with high costs, it is costly that directly enzymolysis process is produced hydrolyzed wheat protein, limit suitability for industrialized production and application, limited especially product applying in some industry.Therefore, the technique of polypeptide is prepared in the microorganism fermentation based on indirect enzyme solution, day by day receives researchist's concern.
Microorganism is the important sources of various zymins.If can screen the microorganism strains that some can directly utilize the protein yield wheat peptide in gluten powder, utilize the metabolic function of microorganism to produce proteolytic enzyme, enzymolysis and these two steps of debitterize are organically combined, by suitable zymotechnique, manufacture wheat peptide cheaply, will have good using value in enormous quantities.Have no at present report.
Summary of the invention
For meeting the need of market, the object of the invention is to provide a kind of novel method that adopts solution fermentation to prepare hydrolyzed wheat protein, enzymolysis and these two steps of debitterize are organically combined, thereby the production cost of reduction hydrolyzed wheat protein is realized suitability for industrialized production.
For realizing the object of the invention, the present invention is according to the feature of the characteristic of wheat gluten flour and animal nutrition, according to microbial fermentation engineering principle and theory, utilize the effect of the microorganism fermentation protein zymins such as the endo-protease that produces, carboxypeptidase that wheat-gluten is hydrolyzed into short peptide chain, and by the hydrophobic amino acid excision of short peptide chain end, realize enzymolysis and debitterize one step completes hydrolyzed wheat protein fermentative Production technique.
Concrete technical scheme is as follows:
1) after aspergillus oryzae (Aspergillu) strain after activation is cultivated through going down to posterity, be transferred in liquid seed culture medium, in 33 ℃, 200r/min shaking culture 36h, is used for fermentation during the little mycelium pellet of homogeneous to be generated; Bacillus subtilis strain (Bacillus subtilis) after activation is inoculated in seed culture medium, 37 ℃, 120r/min shaking culture 23-24h, makes cell concentration reach 10 simultaneously 6-10 8cFU/mL is standby;
2) take wheat-gluten powder, in fermentor tank, adding water, to be configured to weight percent concentration be 5%-10%, add the growing nutrient bag with respect to wheat-gluten quality 0.2%, adjustment pH is 5.5-7.5, then sterilizing is cooled to 33-37 ℃, access above-mentioned subtilis and aspergillus oryzae liquid spawn, bacterial classification weight ratio is: subtilis: aspergillus oryzae=2-5:1, and inoculation total amount is the 5-9 ‰ of system volume;
3) controlled fermentation tank fermentation rotating speed is 50-180rpm, leavening temperature 33-40 ℃, fermentation time 24-40h;
4) after fermentation ends, fermented liquid is carried out to evaporation concentration, then spraying is dried and makes Powdered hydrolyzed wheat protein product.
Described nutrient pack is comprised of glucose, sodium-chlor, potassium primary phosphate, dipotassium hydrogen phosphate, and its weight ratio is 100:1-3:0.5-2:0.1-1.
The present invention utilizes the effect of the microorganism fermentation protein zymins such as the endo-protease that produces, carboxypeptidase that wheat-gluten is hydrolyzed into short peptide chain, and by the hydrophobic amino acid excision of short peptide chain end, realize the hydrolyzed wheat protein fermentative Production technique that enzymolysis and debitterize one step complete.The product of this explained hereafter is to take the deep processing conversion product that agricultural byproducts carry out as raw material, and added value is large, and economic results in society are remarkable.This production technique integrated production capacity is strong, and transformation efficiency is high, reaches more than 80%.Compare with traditional enzyme solution: produce and stablize, product does not have bitter taste, and bitterness value is between 0 and 1, and product special flavour, color and luster are better than enzymatic hydrolysis, and property indices is all better than enzymatic hydrolysis.Molecular weight product is little, and solvability is good, and glutamine content is high, can substitute plasma proteins, and after interpolation application, to improving wean young baby animal digestion rate, solution stress be with the obvious advantage with diarrhoea.Experimental result is as table 1 and table 2:
Table: 1 hydrolyzed wheat protein enzymolysis process and fermentation method result of the present invention contrast
Bitter taste is identified: by the method for Adier-Nissen, using the quinine hydrochloride solution of different concns and as standard, hydrolyzed wheat protein liquid is marked, distilled water is that O divides.
The hydrolyzed wheat protein that table 2 the present invention produces and enzymolysis process product peptide molecular weight distribute and contrast
Note: above data are that sample soluble fractions detects gained molecular weight result
Innovative point of the present invention is: in conjunction with characteristic and the animal nutrition feature of wheat-gluten, utilize the effect of the microorganism fermentation protein zymins such as the endo-protease that produces, carboxypeptidase that wheat-gluten is hydrolyzed into short peptide chain, and by the hydrophobic amino acid excision of short peptide chain end, realize the hydrolyzed wheat protein fermentative Production technique that enzymolysis and debitterize one step complete.Through continuous pilot plant test and scale production checking, realized suitability for industrialized production.Molecular weight product is little, and solvability is good, and glutamine content is high, can substitute plasma proteins, and after interpolation application, to improving wean young baby animal digestion rate, solution stress be with the obvious advantage with diarrhoea, remarkable economic and social benefits, and market outlook are wide.
The hydrolyzed wheat protein product that the present invention produces is compared and is had following features with enzymolysis process: (1) utilizes the subtilis and the conventional safety in production of the aspergillus oryzae bacterial strain mixed fermentation wheat-gluten that filter out to prepare hydrolyzed wheat protein, production technique is advanced, solved direct enzymolysis process production hydrolyzed wheat protein cost large, high in cost of production problem; (2) utilize the metabolic function of microorganism to produce proteolytic enzyme enzymolysis hydrolyzed wheat protein, enzymolysis and these two steps of debitterize are organically combined, solved enzymolysis process product bitter taste large, weak flavor problem; (3) product molecular weight product is little, and solvability is good, and glutamine content is high.
Embodiment
The present invention is further illustrated for the following examples.
Embodiment 1
Aspergillus oryzae strain (Aspergillu) after activation is transferred in liquid seed culture medium, and in 33 ℃, 200r/min shaking culture 36h, is used for fermentation during the little mycelium pellet of homogeneous to be generated; Bacillus subtilis strain (Bacillus subtilis) after activation is inoculated in seed culture medium, 37 ℃, 120r/min shaking culture 24h left and right, makes cell concentration reach 10 simultaneously 6-10 8cFU/mL is standby.Take wheat-gluten powder, in fermentor tank, adding wheat-gluten powder and water to be configured to weight percent concentration is 6%, add the growing nutrient bag with respect to wheat-gluten quality 0.2%, the part by weight of controlling glucose, sodium-chlor, potassium primary phosphate, dipotassium hydrogen phosphate in nutrient pack is 100:1:1:0.5, adjusting pH is 6.0, then sterilizing is cooled to 33 ℃, access subtilis and the aspergillus oryzae liquid spawn of above-mentioned cultivation maturation, bacterial classification part by weight is: subtilis: aspergillus oryzae=2:1, and inoculum size is 5 ‰ of system volume; Controlled fermentation tank fermentation rotating speed is 80rpm, 33 ℃ of leavening temperatures, fermentation time 38h; After fermentation ends, fermented liquid is carried out to evaporation concentration, then spraying is dried and makes Powdered hydrolyzed wheat protein product.
Embodiment 2
Aspergillus oryzae strain (Aspergillu) after activation is transferred in liquid seed culture medium, and in 33 ℃, 200r/min shaking culture 36h, is used for fermentation during the little mycelium pellet of homogeneous to be generated; Bacillus subtilis strain (Bacillus subtilis) after activation is inoculated in seed culture medium, 37 ℃, 120r/min shaking culture 24h left and right, makes cell concentration reach 10 simultaneously 6-10 8cFU/mL is standby.Take wheat-gluten powder, in fermentor tank, adding wheat-gluten powder and water to be configured to weight percent concentration is 7%, add the growing nutrient bag with respect to wheat-gluten quality 0.2%, the part by weight of controlling glucose, sodium-chlor, potassium primary phosphate, dipotassium hydrogen phosphate in nutrient pack is 100:1.5:1.5:0.6, adjusting pH is 6.5, then sterilizing is cooled to 34 ℃, access subtilis and the aspergillus oryzae liquid spawn of above-mentioned cultivation maturation, bacterial classification part by weight is: subtilis: aspergillus oryzae=3:1, and inoculum size is 6 ‰ of system volume; Controlled fermentation tank fermentation rotating speed is 120rpm, 34 ℃ of leavening temperatures, fermentation time 40h; After fermentation ends, fermented liquid is carried out to evaporation concentration, then spraying is dried and makes Powdered hydrolyzed wheat protein product.
Embodiment 3
Aspergillus oryzae strain (Aspergillu) after activation is transferred in liquid seed culture medium, and in 33 ℃, 200r/min shaking culture 36h, is used for fermentation during the little mycelium pellet of homogeneous to be generated; Bacillus subtilis strain (Bacillus subtilis) after activation is inoculated in seed culture medium, 37 ℃, 120r/min shaking culture 24h left and right, makes cell concentration reach 10 simultaneously 6-10 8cFU/mL is standby.Take wheat-gluten powder, in fermentor tank, adding wheat-gluten powder and water to be configured to weight percent concentration is 8%, add the growing nutrient bag with respect to wheat-gluten quality 0.2%, the part by weight of controlling glucose, sodium-chlor, potassium primary phosphate, dipotassium hydrogen phosphate in nutrient pack is 100:2:1:0.3, adjusting pH is 7.0, then sterilizing is cooled to 35 ℃, access subtilis and the aspergillus oryzae liquid spawn of above-mentioned cultivation maturation, bacterial classification part by weight is: subtilis: aspergillus oryzae=5:1, and inoculum size is 7 ‰ of system volume; Controlled fermentation tank fermentation rotating speed is 160rpm, 35 ℃ of leavening temperatures, fermentation time 38h; After fermentation ends, fermented liquid is carried out to evaporation concentration, then spraying is dried and makes Powdered hydrolyzed wheat protein product.
Embodiment 4
Aspergillus oryzae strain (Aspergillu) after activation is transferred in liquid seed culture medium, and in 33 ℃, 200r/min shaking culture 36h, can be used for fermentation during the little mycelium pellet of homogeneous to be generated; Bacillus subtilis strain (Bacillus subtilis) after activation is inoculated in seed culture medium, 37 ℃, 120r/min shaking culture 24h left and right, makes cell concentration reach 10 simultaneously 6-10 8cFU/mL is standby.Take wheat-gluten powder, in fermentor tank, adding wheat-gluten powder and water to be configured to weight percent concentration is 10%, add the quality 0.2% growing nutrient bag with respect to wheat-gluten, the part by weight of controlling glucose, sodium-chlor, potassium primary phosphate, dipotassium hydrogen phosphate in nutrient pack is 100:3:2:1, adjusting pH is 5.6, then sterilizing is cooled to 33 ℃, access subtilis and the aspergillus oryzae liquid spawn of above-mentioned cultivation maturation, bacterial classification part by weight is: subtilis: aspergillus oryzae=4:1, and inoculum size is 8 ‰ of system volume; Controlled fermentation tank fermentation rotating speed is 50rpm, 33 ℃ of leavening temperatures, fermentation time 40h; After fermentation ends, fermented liquid is carried out to evaporation concentration, then spraying is dried and makes Powdered hydrolyzed wheat protein product.

Claims (1)

1. fermentation method is prepared a production method for hydrolyzed wheat protein, it is characterized in that, by following steps, is realized:
(1) aspergillus oryzae strain (Aspergillu) after activation is transferred in liquid seed culture medium, in 33 ℃, 200r/min shaking culture 36h, is used for fermentation during the little mycelium pellet of homogeneous to be generated; Bacillus subtilis strain (Bacillus subtilis) after activation is inoculated in seed culture medium, 37 ℃, 120r/min shaking culture 23-24h, makes cell concentration reach 10 simultaneously 6-10 8cFU/mL is standby; (2) take wheat-gluten powder, in fermentor tank, adding water, to be configured to weight percent concentration be 5%-10%, add the growing nutrient bag with respect to wheat-gluten quality 0.2%, adjustment pH is 5.5-7.5, then sterilizing is cooled to 33-37 ℃, access above-mentioned subtilis and aspergillus oryzae liquid spawn, bacterial classification weight ratio is: subtilis: aspergillus oryzae=2-5:1, and inoculation total amount is the 5-9 ‰ of system volume; (3) controlled fermentation tank fermentation rotating speed is 50-180rpm, leavening temperature 33-40 ℃, fermentation time 24-40h; (4) after fermentation ends, fermented liquid is carried out to evaporation concentration, then spraying is dried and makes Powdered hydrolyzed wheat protein product;
Described growing nutrient bag is comprised of glucose, sodium-chlor, potassium primary phosphate, dipotassium hydrogen phosphate, and its weight ratio is 100:1-3:0.5-2:0.1-1.
CN201410533095.3A 2014-10-11 2014-10-11 Production method for preparing hydrolyzed wheat proteins by fermentation method Pending CN104212859A (en)

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Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105230987A (en) * 2015-11-02 2016-01-13 山东贝瑞康生物科技有限公司 Healthcare agent for pet puppies
CN105410347A (en) * 2015-11-02 2016-03-23 山东贝瑞康生物科技有限公司 Health care agent for old dogs

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1840673A (en) * 2006-01-19 2006-10-04 江南大学 Process for preparing feruoylated oligosaccharide by enzymolysis of wheat bran
CN101156645A (en) * 2007-11-06 2008-04-09 江苏大学 A wheat gluten proteolysis matter and method for making same as well as its usage
CN101627793A (en) * 2009-07-31 2010-01-20 郑州新威营养技术有限公司 Industrial production method of hydrolyzed wheat protein for feeding
CN102907559A (en) * 2012-11-15 2013-02-06 温志明 Process for industrial production of hydrolyzed wheat protein
CN103907748A (en) * 2014-04-04 2014-07-09 烟台金富基生物科技有限公司 Method for preparing multi-nutrient biological peptide

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN1840673A (en) * 2006-01-19 2006-10-04 江南大学 Process for preparing feruoylated oligosaccharide by enzymolysis of wheat bran
CN101156645A (en) * 2007-11-06 2008-04-09 江苏大学 A wheat gluten proteolysis matter and method for making same as well as its usage
CN101627793A (en) * 2009-07-31 2010-01-20 郑州新威营养技术有限公司 Industrial production method of hydrolyzed wheat protein for feeding
CN102907559A (en) * 2012-11-15 2013-02-06 温志明 Process for industrial production of hydrolyzed wheat protein
CN103907748A (en) * 2014-04-04 2014-07-09 烟台金富基生物科技有限公司 Method for preparing multi-nutrient biological peptide

Cited By (2)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN105230987A (en) * 2015-11-02 2016-01-13 山东贝瑞康生物科技有限公司 Healthcare agent for pet puppies
CN105410347A (en) * 2015-11-02 2016-03-23 山东贝瑞康生物科技有限公司 Health care agent for old dogs

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