CN107312813A - A kind of preparation method of microorganism specialized soybean protein isolate - Google Patents
A kind of preparation method of microorganism specialized soybean protein isolate Download PDFInfo
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- 108010073771 Soybean Proteins Proteins 0.000 title claims abstract description 60
- 235000019710 soybean protein Nutrition 0.000 title claims abstract description 60
- 244000005700 microbiome Species 0.000 title claims abstract description 48
- 238000002360 preparation method Methods 0.000 title claims abstract description 32
- 239000002253 acid Substances 0.000 claims abstract description 75
- 239000007788 liquid Substances 0.000 claims abstract description 60
- 235000018102 proteins Nutrition 0.000 claims abstract description 37
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 37
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- 238000000034 method Methods 0.000 claims abstract description 35
- 239000003513 alkali Substances 0.000 claims abstract description 30
- 108091005804 Peptidases Proteins 0.000 claims abstract description 22
- 239000004365 Protease Substances 0.000 claims abstract description 22
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 22
- 235000019419 proteases Nutrition 0.000 claims abstract description 22
- 101000693530 Staphylococcus aureus Staphylokinase Proteins 0.000 claims abstract description 20
- 235000019764 Soybean Meal Nutrition 0.000 claims abstract description 19
- 239000000287 crude extract Substances 0.000 claims abstract description 19
- 238000007701 flash-distillation Methods 0.000 claims abstract description 19
- 239000004455 soybean meal Substances 0.000 claims abstract description 19
- 238000003756 stirring Methods 0.000 claims abstract description 19
- 230000001954 sterilising effect Effects 0.000 claims abstract description 17
- 238000004659 sterilization and disinfection Methods 0.000 claims abstract description 14
- 238000000926 separation method Methods 0.000 claims abstract description 13
- 238000001694 spray drying Methods 0.000 claims abstract description 12
- 239000007787 solid Substances 0.000 claims abstract description 8
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- 239000000243 solution Substances 0.000 claims description 26
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 21
- 239000007864 aqueous solution Substances 0.000 claims description 21
- 238000005507 spraying Methods 0.000 claims description 9
- 230000001939 inductive effect Effects 0.000 claims description 6
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- 235000010469 Glycine max Nutrition 0.000 claims description 4
- NBIIXXVUZAFLBC-UHFFFAOYSA-N Phosphoric acid Chemical compound OP(O)(O)=O NBIIXXVUZAFLBC-UHFFFAOYSA-N 0.000 claims description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 claims description 4
- 244000068988 Glycine max Species 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
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- 229910052757 nitrogen Inorganic materials 0.000 abstract description 19
- 150000001413 amino acids Chemical class 0.000 abstract description 16
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- 239000006185 dispersion Substances 0.000 abstract 1
- 230000001737 promoting effect Effects 0.000 abstract 1
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 10
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- 206010000496 acne Diseases 0.000 description 7
- 238000012360 testing method Methods 0.000 description 7
- 230000009286 beneficial effect Effects 0.000 description 6
- 230000000052 comparative effect Effects 0.000 description 6
- 239000007921 spray Substances 0.000 description 6
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 5
- 229910052799 carbon Inorganic materials 0.000 description 5
- 230000003750 conditioning effect Effects 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
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- 239000000843 powder Substances 0.000 description 4
- 230000017854 proteolysis Effects 0.000 description 4
- 108090000145 Bacillolysin Proteins 0.000 description 3
- NIPNSKYNPDTRPC-UHFFFAOYSA-N N-[2-oxo-2-(2,4,6,7-tetrahydrotriazolo[4,5-c]pyridin-5-yl)ethyl]-2-[[3-(trifluoromethoxy)phenyl]methylamino]pyrimidine-5-carboxamide Chemical compound O=C(CNC(=O)C=1C=NC(=NC=1)NCC1=CC(=CC=C1)OC(F)(F)F)N1CC2=C(CC1)NN=N2 NIPNSKYNPDTRPC-UHFFFAOYSA-N 0.000 description 3
- 102000035092 Neutral proteases Human genes 0.000 description 3
- 108091005507 Neutral proteases Proteins 0.000 description 3
- 239000001888 Peptone Substances 0.000 description 3
- 108010080698 Peptones Proteins 0.000 description 3
- 244000046052 Phaseolus vulgaris Species 0.000 description 3
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 3
- 239000003153 chemical reaction reagent Substances 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- 239000001963 growth medium Substances 0.000 description 3
- 235000019319 peptone Nutrition 0.000 description 3
- 238000012545 processing Methods 0.000 description 3
- 150000003839 salts Chemical class 0.000 description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 2
- 240000004808 Saccharomyces cerevisiae Species 0.000 description 2
- 239000007640 basal medium Substances 0.000 description 2
- 238000004090 dissolution Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 238000000605 extraction Methods 0.000 description 2
- 239000000706 filtrate Substances 0.000 description 2
- 238000001914 filtration Methods 0.000 description 2
- 238000001879 gelation Methods 0.000 description 2
- 239000000463 material Substances 0.000 description 2
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- 239000012898 sample dilution Substances 0.000 description 2
- 239000012086 standard solution Substances 0.000 description 2
- 238000010998 test method Methods 0.000 description 2
- JLPULHDHAOZNQI-ZTIMHPMXSA-N 1-hexadecanoyl-2-(9Z,12Z-octadecadienoyl)-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCCCCCC\C=C/C\C=C/CCCCC JLPULHDHAOZNQI-ZTIMHPMXSA-N 0.000 description 1
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 1
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 1
- 101710089165 Protein white Proteins 0.000 description 1
- NINIDFKCEFEMDL-UHFFFAOYSA-N Sulfur Chemical compound [S] NINIDFKCEFEMDL-UHFFFAOYSA-N 0.000 description 1
- 239000005864 Sulphur Substances 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 230000032683 aging Effects 0.000 description 1
- 235000004458 antinutrient Nutrition 0.000 description 1
- 241000758522 bacterium 20S Species 0.000 description 1
- 235000004251 balanced diet Nutrition 0.000 description 1
- 239000002585 base Substances 0.000 description 1
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- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000012141 concentrate Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000001816 cooling Methods 0.000 description 1
- 239000013530 defoamer Substances 0.000 description 1
- 238000007872 degassing Methods 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 230000000694 effects Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000019634 flavors Nutrition 0.000 description 1
- 239000012530 fluid Substances 0.000 description 1
- 239000011521 glass Substances 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 230000007062 hydrolysis Effects 0.000 description 1
- 238000006460 hydrolysis reaction Methods 0.000 description 1
- 238000005213 imbibition Methods 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 238000003760 magnetic stirring Methods 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000002906 microbiologic effect Effects 0.000 description 1
- 235000013336 milk Nutrition 0.000 description 1
- 239000008267 milk Substances 0.000 description 1
- 210000004080 milk Anatomy 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 150000004965 peroxy acids Chemical class 0.000 description 1
- 229910052698 phosphorus Inorganic materials 0.000 description 1
- 239000011574 phosphorus Substances 0.000 description 1
- 238000001556 precipitation Methods 0.000 description 1
- 230000003946 protein process Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 210000000697 sensory organ Anatomy 0.000 description 1
- 239000010865 sewage Substances 0.000 description 1
- 229940083466 soybean lecithin Drugs 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000012549 training Methods 0.000 description 1
- 239000003643 water by type Substances 0.000 description 1
- 230000010148 water-pollination Effects 0.000 description 1
- 210000004885 white matter Anatomy 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
Classifications
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- General Health & Medical Sciences (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- General Engineering & Computer Science (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Botany (AREA)
- Gastroenterology & Hepatology (AREA)
- Biophysics (AREA)
- Medicinal Chemistry (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：At a temperature of 50 55 DEG C, defatted soybean meal is dipped in the aqueous slkali that pH value is 9.5~10.0, regulation pH value is 6.8~7.4, pol is 8.0~9.0, stirs 40 60min；Through separation of solid and liquid, protein crude extract is obtained；At room temperature, acid solution is added into above-mentioned protein crude extract, 10 50min, the centrifuged heavy albumen of acid are stirred；At room temperature, it is the heavy albumen of above-mentioned acid is soluble in water, pH to 6.00~6.30 is adjusted, pol is 18.0~19.5, obtains sour heavy protein liquid；Neutral proteinase and alkali protease are added, at 50 60 DEG C, 0.5 3h is reacted, obtains enzymolysis liquid；Through sterilization, flash distillation process, spray drying is produced.Soybean protein isolate good dispersion prepared by the present invention, viscosity are low, amino-acid nitrogen is suitable, and technique is simple, easily operated, and albumen is practical, is widely used in fermentation industry, with preferable promotional value.
Description
Technical field
The present invention relates to a kind of preparation method of microorganism specialized soybean protein isolate, belong to vegetable protein process technology
Field.
Background technology
Soybean protein isolate contains abundant carbon source and nitrogen source, and containing eight kinds of necessary amino acid of human body, biological value is high,
It is easy to digest and assimilate.Soybean protein isolate product is nutritious, reasonable, and structure preferably, plays an important role to balanced diet,
Had broad application prospects in food industry.Soybean protein isolate not only yields unusually brilliant results in food industry, in microbial fermentation
Industry also has a wide range of applications.
At present, because soybean protein isolate has good dissolubility, emulsibility and gelation and is widely used in food
Multiple fields, practicality is stronger, but to seldom, the special soybean separation of no culture microorganism of research in terms of microculture
Albumen.Soybean protein isolate prepared by prior art, although meeting the demand of microorganism growth, molecular weight is high, is not suitable for micro-
Biological digests and assimilates, grows utilization；Plain edition soybean protein isolate is dissolved in after water, and viscosity is too high, is unfavorable for cultivating microorganism
Scale, serialization and industrialization；Plain edition soybean protein isolate is more in the pimple that whipping process is formed, operating difficulties,
Requirement to equipment is harsh；And amino-acid nitrogen is not suitable for, carbon source is too high, and microbial fermentation solution is partially yellow, the easy aging of mycelia, spore
Son is more；When nitrogen source is too high, mycelia is elongated, and metabolite is more difficult to be extracted, and disadvantages described above have impact on the growth situation of microorganism,
Influence the quality of product.Disadvantages described above limits application of the soybean protein isolate in terms of microbial fermentation.
The A of Chinese patent literature CN 101642184 disclose a kind of injection-type soybean protein isolate and preparation method thereof, adopt
It is raw material with low temperature soy meal, by alkali carries, degassing, the heavy preparative separation albumen curd of acid, then using neutral proteinase to separation
Albumen curd is digested, and is added after enzymolysis in enzymolysis product and is puddled and use soybean lecithin, after gone out enzyme, sterilizing, sudden strain of a muscle by heating
Steaming, homogeneous, spray drying treatment obtain all preferable soybean protein of dispersiveness, water imbibition, water-retaining property, oil absorption and gelation.
But the invention is only with neutral protein ferment treatment and feed liquid, and usage amount is less, to proteolysis limitation, amino acid
State nitrogen is not suitable for, and is unfavorable for the extraction of microbial metabolic products.
The A of Chinese patent literature CN 101372501 disclose a kind of preparation method of soybean protein isolate, including following step
Suddenly：(1) it is dried in vacuo the preparation of alcohol processed soybean protein concentrate powder；(2) solution is prepared and is heat-treated；(3) acid is heavy adjusts neutral；(4)
Instantaneous heat sterilization and spray drying.Prepared soybean protein isolate had both had highly dissoluble, the high egg of common separation albumen
Bai Hanliang advantage, again have beany flavor it is low, lighter color, anti-nutrient substance activity is low, sewage effluent is easily handled wait commonly it is big
The advantage that beans protein isolate does not possess.But the preparation method sterilization temperature is relatively low, and the viscosity that may result in product is too high, dissolving
The pimple of process formation is more, is not suitable for the culture as microorganism.
The present invention is proposed to solve the above problems.
The content of the invention
The present invention provides simple, easy in a kind of preparation method of microorganism specialized soybean protein isolate, operation sequence,
Prepared soybean protein isolate disclosure satisfy that the demand of microorganism growth, and molecular weight is suitable, and carbon source and nitrogen source are suitable, are dissolved in water
There is relatively low viscosity afterwards, pimple is less, beneficial to the scale of microorganism, serialization and industrialization culture.
Technical scheme is as follows：
Term explanation：
Curdled milk is dehydrated：After finger protein crude extract sinks through peracid, dehydration is centrifuged, the heavy albumen of obtained precipitation i.e. acid
Water content.
Spraying ratio：The ratio of the quality for referring to spray and the quality for being sprayed-on thing.
A kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At a temperature of 50-55 DEG C, defatted soybean meal is dipped in the aqueous slkali that pH value is 9.5~10.0, adjusted
It is 6.8~7.4 to save pH value, and pol is 8.0~9.0, stirs 40-60min；Through separation of solid and liquid, protein crude extract is obtained；
(2) acid is heavy：At room temperature, acid solution is added in the protein crude extract obtained to step (1), 10-50min, warp is stirred
Centrifugation obtains the heavy albumen of acid；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, adjusts pH to 6.00~6.30,
Pol is 18.0~19.5, obtains sour heavy protein liquid；Neutral proteinase and alkali protease are added, at 50-60 DEG C, 0.5- is reacted
3h, obtains enzymolysis liquid；
(4) refine：The enzymolysis liquid that step (3) is obtained produces microorganism tailored version through sterilization, flash distillation process, spray drying
Soybean protein isolate.
According to currently preferred, the mass ratio of defatted soybean meal and aqueous slkali described in step (1) is 1:7-11.
It is preferred that, the mass ratio of defatted soybean meal and aqueous slkali described in step (1) is 1:9.5.
According to currently preferred, aqueous slkali described in step (1) is the NaOH or KOH aqueous solution.
According to currently preferred, pH is adjusted using acid solution in step (1), the mass concentration of the acid solution is 30-
35%.
According to the present invention, pol is adjusted using process water or defatted soybean meal through reverse-osmosis treated.
It is preferred that, the acid solution is the aqueous solution of hydrochloric acid, sulfuric acid, phosphoric acid or glacial acetic acid.
According to currently preferred, the acid solution described in step (2) is that the hydrochloric acid that mass concentration is 20-40% is water-soluble
Liquid, the pH of regulatory protein crude extract is 4.50~4.6；The heavy albumen curd dehydration 50~56% of acid described in step (2).
According to currently preferred, the NaOH for being 20-40% with mass concentration in step (3) or KOH aqueous solution tune pH.
According to currently preferred, the mass concentration of the heavy albumen of acid is 40-50% described in the heavy protein liquid of step (3) acid.
According to currently preferred, the mass ratio of acid described in step (3) heavy albumen, neutral proteinase and alkali protease
For 100：0.8~1.2：1.4~1.8.
According to currently preferred, method for disinfection is described in step (4)：It is 145~165 DEG C, vacuum in sterilization temperature
Spend to be sterilized twice under the conditions of -0.005~0.03Mpa, sterilizing time is respectively 10-30s and 40-90s, described to kill twice
Bacterium temperature and vacuum all same.
According to currently preferred, flash conditions are described in step (4)：Flash distillation liquid level is 45-55%, flash vacuum degree
For -0.005~-0.015Mpa.
According to currently preferred, spray drying condition is described in step (4)：Wind pushing temperature is 190~220 DEG C, is drawn
Air temperature is 80~90 DEG C, is sprayed using liquid phosphatide, and the spraying ratio of liquid phosphatide is 4 ‰.
Beneficial effects of the present invention are as follows：
(1) present invention uses neutral proteinase and alkali protease joint enzymolysis, suitable to proteolysis degree, obtains
The molecular weight of the soybean protein isolate arrived is suitable, beneficial to the digesting and assimilating of microorganism, grows utilization；It is dissolved in after water, viscosity is suitable,
Beneficial to the scale of microorganism, serialization and industrialization culture；With suitable amino-acid nitrogen and carbon source, beneficial to microorganism generation
Thank to the extraction of product.Neutral proteinase and alkali protease mixing combination efficiency high, can control the degree of proteolysis, simultaneously
Without needing as the processing of peptone, proteolysis extent control to high, processing cost is reduced.
(2) present invention is effectively sterilized using high-temperature sterilization technique twice, reduce further soybean protein isolate
Molecular weight, reduce soybean protein isolate and be dissolved in the viscosity after water, while also reducing the pimple number formed in course of dissolution
Amount, and the content of amino-acid nitrogen is further reduced, make with amino acid nitrogen content preferably, and effectively increase big
The dispersiveness of beans protein isolate；Carrying out spraying using liquid phosphatide simultaneously improves the hydrophily of product.
(3) preparation method of the present invention is simple, it is easy to operate, prepared microorganism specialized soybean protein isolate dispersiveness
Well, molecular weight is suitable, viscosity is low, amino-acid nitrogen and carbon source are suitable, and the microorganism growth situation of culture is good, microorganism
Product yield high；The protein content of prepared microorganism specialized soybean protein isolate up to more than 90%, viscosity 60~
150cps, amino-acid nitrogen 0.32~0.45% meets the composition of microorganism growth, and it is excellent to be adapted to Continuous maching etc.
Point, can be widely applied in food fermentation industry.
Embodiment
With reference to specific embodiment, the present invention is described further, but not limited to this.
Experimental method described in following embodiments, is conventional method unless otherwise specified simultaneously；The reagent and material
Material, unless otherwise specified, is commercially obtained.
In embodiment, defatted soybean meal, Shandong king Yu ecology Industrial Co., Ltd. is on sale；Neutral proteinase is Denis
Gram neutral proteinase；Alkali protease is Danisco food-grade alkali protease.
Embodiment 1
A kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At a temperature of 50-55 DEG C, defatted soybean meal is dipped in the NaOH aqueous solution that pH value is 9.6, it is described
The mass ratio of defatted soybean meal and the NaOH aqueous solution is 1:9.5, the salt acid for adjusting pH value for being 30-35% with mass concentration is
6.8, it is 8.2 with the process water regulation pol through reverse-osmosis treated, stirs 50min；Through separation of solid and liquid, protein crude extract is obtained；
(2) acid is heavy：At room temperature, the pH of 30% hydrochloric acid conditioning solution is added in the protein crude extract obtained to step (1)
For 4.54, stirring 20min, the centrifuged heavy albumen of acid, wherein the heavy albumen curd of acid is dehydrated 52%；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, is 30% with mass concentration
The NaOH aqueous solution adjusts pH to 6.18, adjusts pol to be 18.6 with the process water through reverse-osmosis treated, obtains sour heavy protein liquid；Add neutral
Protease and alkali protease, at 55 DEG C, react 1h, obtain enzymolysis liquid；The quality of the heavy albumen of acid is dense in the heavy protein liquid of acid
Spend for 48%, the mass ratio of the acid heavy albumen, neutral proteinase and alkali protease is 100:1:1.5.
(4) refine：The enzymolysis liquid that step (3) is obtained temperature be 158 DEG C, vacuum be -0.005Mpa under the conditions of, respectively
Sterilize 15s and 60s；Through flash distillation process, spray drying prepares microorganism specialized soybean protein isolate.The flash distillation process
Condition is；Vacuum is -0.005Mpa, and flash distillation liquid level is 45-55%；The spray condition is：Wind pushing temperature：198 DEG C, air inducing
Temperature is 84 DEG C, is sprayed using liquid phosphatide, and spraying ratio is 4 ‰.
Embodiment 2
A kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At a temperature of 50-55 DEG C, defatted soybean meal is dipped in the KOH aqueous solution that pH value is 9.8, it is described low
The mass ratio of warm defatted soybean meal and the KOH aqueous solution is 1:9.5, the salt acid for adjusting pH value for being 30-35% with mass concentration is 6.8,
It is 8.6 with the process water regulation pol through reverse-osmosis treated, stirs 50min；Through separation of solid and liquid, protein crude extract is obtained；
(2) acid is heavy：At room temperature, the pH of 30% hydrochloric acid conditioning solution is added in the protein crude extract obtained to step (1)
For 4.52, stirring 20min, the centrifuged heavy albumen of acid, wherein the heavy albumen curd of acid is dehydrated 51.6%；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, is 30% with mass concentration
The NaOH aqueous solution adjusts pH to 6.21, adjusts pol to be 19.1 with the process water through reverse-osmosis treated, obtains sour heavy protein liquid；Add neutral
Protease and alkali protease, at 50 DEG C, react 1h, obtain enzymolysis liquid；The quality of the heavy albumen of acid is dense in the heavy protein liquid of acid
Spend for 48%, the mass ratio of the acid heavy albumen, neutral proteinase and alkali protease is 100:0.8:1.7.
(4) refine：The enzymolysis liquid that step (3) is obtained temperature be 156 DEG C, vacuum be 0.03Mpa under the conditions of, kill respectively
Bacterium 15s and 60s；Through flash distillation process, spray drying prepares microorganism specialized soybean protein isolate.The flash distillation process bar
Part is；Vacuum is -0.006Mpa, and flash distillation liquid level is 45-55%；The spray condition is：Wind pushing temperature：196 DEG C, air inducing temperature
Spend for 85 DEG C, sprayed using liquid phosphatide, spraying ratio is 4 ‰.
Embodiment 3
A kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At 50-55 DEG C, defatted soybean meal is dipped in the KOH aqueous solution that pH value is 9.6, the low temperature takes off
The mass ratio of fat dregs of beans and the KOH aqueous solution is 1:9.5, the salt acid for adjusting pH value for being 30-35% with mass concentration is 7, with through anti-
The process water regulation pol of infiltration processing is 8.2, stirs 50min；Through separation of solid and liquid, protein crude extract is obtained；
(2) acid is heavy：At room temperature, the pH of 30% hydrochloric acid conditioning solution is added in the protein crude extract obtained to step (1)
For 4.55, stirring 20min, the centrifuged heavy albumen of acid, wherein the heavy albumen curd of acid is dehydrated 51.7%；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, is 30% with mass concentration
The NaOH aqueous solution adjusts pH to 6.28, adjusts pol to be 19.5 with the process water through reverse-osmosis treated, obtains sour heavy protein liquid；Add neutral
Protease and alkali protease, at 53 DEG C, react 1h, obtain enzymolysis liquid；The quality of the heavy albumen of acid is dense in the heavy protein liquid of acid
Spend for 48%, the mass ratio of the acid heavy albumen, neutral proteinase and alkali protease is 100:0.9:1.6.
(4) refine：The enzymolysis liquid that step (3) is obtained temperature be 160 DEG C, vacuum be 0.01Mpa under the conditions of, kill respectively
Bacterium 15s and 60s；Through flash distillation process, spray drying prepares microorganism specialized soybean protein isolate.The flash distillation process bar
Part is；Vacuum is under -0.005Mpa, flash distillation liquid level is 45-55%；The spray condition is：Wind pushing temperature：199 DEG C, air inducing
Temperature is 86 DEG C, is sprayed using liquid phosphatide, and spraying ratio is 4 ‰.
Embodiment 4
A kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At 50-55 DEG C, defatted soybean meal is dipped in the NaOH aqueous solution that pH value is 9.6, the low temperature
The mass ratio of defatted soybean meal and the NaOH aqueous solution is 1:7, the sulphur acid for adjusting pH value for being 30-35% with mass concentration is 7.4, with warp
The process water regulation pol of reverse-osmosis treated is 8.0, stirs 50min；Through separation of solid and liquid, protein crude extract is obtained；
(2) acid is heavy：At room temperature, the pH of 30% hydrochloric acid conditioning solution is added in the protein crude extract obtained to step (1)
For 4.54, stirring 20min, the centrifuged heavy albumen of acid, wherein the heavy albumen curd of acid is dehydrated 52%；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, is 30% with mass concentration
The NaOH aqueous solution adjusts pH to 6.18, adjusts pol to be 18.6 with the process water through reverse-osmosis treated and the heavy albumen mixing of acid, obtains sour heavy
Protein liquid；Neutral proteinase and alkali protease are added, at a temperature of 50 DEG C, 0.5h is reacted, obtains enzymolysis liquid；The heavy albumen of acid
The mass concentration of the heavy albumen of acid is 48% in liquid, and the mass ratio of the acid heavy albumen, neutral proteinase and alkali protease is
100:1.2:1.4。
(4) refine：The enzymolysis liquid that step (3) is obtained temperature be 155 DEG C, vacuum be 0.01Mpa under the conditions of, kill respectively
Bacterium 10s and 50s；Through flash distillation process, spray drying prepares microorganism specialized soybean protein isolate.The flash distillation process bar
Part is；Under vacuum is -0.01Mpa, liquid level 45-55% is flashed；The spray condition is：Wind pushing temperature：190 DEG C, air inducing temperature
Spend for 80 DEG C, sprayed using liquid phosphatide, spraying ratio is 4 ‰.
Embodiment 5
A kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At 50-55 DEG C, defatted soybean meal is dipped in the NaOH aqueous solution that pH value is 9.6, the low temperature
The mass ratio of defatted soybean meal and the NaOH aqueous solution is 1:11, the phosphorus acid for adjusting pH value for being 30-35% with mass concentration is 6.8, is used
Process water and defatted soybean meal mixed liquor regulation pol through reverse-osmosis treated are 9.0, stir 50min；Through separation of solid and liquid, obtain
To protein crude extract；
(2) acid is heavy：At room temperature, the pH of 30% hydrochloric acid conditioning solution is added in the protein crude extract obtained to step (1)
For 4.6, stirring 20min, the centrifuged heavy albumen of acid, wherein the heavy albumen curd of acid is dehydrated 52%；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, is 30% with mass concentration
The NaOH aqueous solution adjusts pH to 6.3, is 18.0 with the process water through reverse-osmosis treated and the heavy albumen mixing regulation pol of acid, obtains sour heavy
Protein liquid；Neutral proteinase and alkali protease are added, at a temperature of 60 DEG C, 3h is reacted, obtains enzymolysis liquid；The heavy protein liquid of acid
The mass concentration of the middle heavy albumen of acid is 48%, and the mass ratio of the acid heavy albumen, neutral proteinase and alkali protease is 100:
1.2:1.8。
(4) refine：The enzymolysis liquid that step (3) is obtained temperature be 160 DEG C, vacuum be 0.01Mpa under the conditions of, kill respectively
Bacterium 20s and 80s；Through flash distillation process, spray drying prepares microorganism specialized soybean protein isolate.The flash distillation process bar
Part is；Vacuum is under -0.01Mpa, flash distillation liquid level is 45-55%；The spray condition is：Wind pushing temperature：200 DEG C, air inducing
Temperature is 90 DEG C, is sprayed using liquid phosphatide, and spraying ratio is 4 ‰.
Comparative example 1
A kind of preparation method of soybean protein isolate, preparation process as different from Example 1, is only added in step (3)
Alkali protease, the mass ratio of the heavy albumen of acid and alkali protease is 100:2.5, other steps are consistent.
Comparative example 2
A kind of preparation method of soybean protein isolate, preparation process as different from Example 1, sterilization side in step (4)
Method is：Temperature be 158 DEG C, vacuum be -0.005Mpa under, once sterilized, sterilizing time is 60s, other steps one
Cause.
Comparative example 3
A kind of preparation method of soybean protein isolate, preparation process as different from Example 1, is only added in step (3)
Neutral proteinase, the mass ratio of the heavy albumen of acid and neutral proteinase is 100:2.5, other steps are consistent.
Test example 1
Sense organ, physics and chemistry and microorganism testing are carried out to soybean protein isolate prepared by embodiment 1-5 and comparative example 1-3,
Method of testing is as follows：
1st, the measure of amino acid nitrogen content：Amino-acid nitrogen is measured using formolite number method.5.0g samples are weighed to put
In 100mL volumetric flask, draw 20mL after being diluted with water to scale, mixing, be placed in 200mL beaker, plus 60mL water,
Turn on agitator, acidometer is titrated to standard solution of sodium hydroxide (c (NaOH)=0.05mol/L) and indicates that pH is 8.2, is write down
The milliliter number of Standard Volumetric Solutions for Sodium Hydroxide is consumed, total acid content can be calculated.10mL formalin is added, is mixed.Use again
Standard solution of sodium hydroxide titrates pH to 9.2, writes down the milliliter number of consumption Standard Volumetric Solutions for Sodium Hydroxide.Take 80mL's simultaneously
Water, it is 8.2 first to be adjusted with sodium hydroxide to pH, adds 10mL formalin, is titrated with Standard Volumetric Solutions for Sodium Hydroxide
PH is to 9.2, while doing reagent blank experiment.
X={ (V1-V2)×C×0.014/5×(V3/100)}×100
X：The content of amino-acid nitrogen in sample, unit is gram every hectogram；
V1：Determine the volume that formaldehyde post consumption Standard Volumetric Solutions for Sodium Hydroxide is added with sample dilution；
V2：Reagent blank experiment adds the volume of formaldehyde post consumption Standard Volumetric Solutions for Sodium Hydroxide；
V3：The sample dilution amount of taking.
2nd, the measure of viscosity：0~5 DEG C of distilled water for measuring 240mL is poured into agitator, is accurately weighed with precision balance
60.0g soybean protein isolates, are poured into agitator, and agitator is adjusted into 1 grade, stir 10s, and clear wall stirs 30s, and clear wall continues
Stirring 30s after pour into 250mL beakers in tall form, stand 1h after, plus 2 drop defoamer after be measured, location parameter is：No. 3 turn
Son, 40 rotating speeds, termination condition 10s.
3rd, the measure of pimple weight：10.0g soybean protein isolate is weighed, is dissolved in 40.0g process water, 1min is stirred,
It is stirred with the speed that each second 2 encloses, crosses the sieve of 40 mesh, weigh the weight of oversize.
4th, the measure of protein content：Accurately weigh sample 5.0g to be placed in 250mL beakers, added into 250mL beakers
100mL distilled water added 100mL distilled water in stirring at low speed on magnetic stirring apparatus 1 hour, and 40 DEG C of keeping temperature continues
Stirring 1 hour, the extract that stirring is finished is transferred in 250mL volumetric flask, and constant volume after cooling fully shakes up constant volume liquid
Pour into centrifuge tube, centrifuged 10 minutes in the centrifuge of 4000 revs/min of rotating speed, centrifugation is finished, by centrifugate fast grade filter paper
Filtering, collects filtrate in conical flask.Draw 50mL filtrates to be placed in 250mL beakers, stand 30 minutes, then be placed in centrifuge tube
Centrifuged 10 minutes in the centrifuge of 4000 revs/min of rotating speed, by supernatant liquid filtering, take 50mL filtered fluids to carry out with 50mL pipettes
Determining the protein quantity, protein content is measured using the method for Kai Shiding nitrogen, and it is multiplied by coefficient 6.25 is converted to thick egg
The content of white matter.Hydrolysis degree={ (V1-V2)×C×0.014×6.25×10×100×100/M×(100-W)}×100
In formula：V2：The volume (mL) of quota of expenditure HCl solution when titrating absorbing liquid；
V1：The volume (mL) of quota of expenditure HCl solution when titrating blank；
C：Standard HCl solution molar concentration；
K：Protein factor 6.25；
W：The water content of albumen powder；
M：Albumen powder quality (g).
5th, dispersiveness test：Measure 190mL room temperature waters to be put into 500mL beakers, then weigh 10g samples and be put in beaker,
It is sufficiently stirred for glass bar, observes the fully decentralized time.
Test result is as shown in table 1：
Table 1
There is high albumen to contain for comprehensive analysis, as shown in Table 1, microorganism specialized soybean protein isolate prepared by the present invention
Amount, it is smaller to be dissolved in viscosity after water, and preferably, amino acid nitrogen content is suitable, and pimple quantity is few, content of microorganisms after sterilizing for dispersiveness
It is low.The present invention uses neutral proteinase and alkali protease joint enzymolysis, effectively reduces soybean protein isolate and is dissolved in
Viscosity after water, makes amino acid nitrogen content suitable；Using high-temperature sterilization technique twice, effectively sterilized, further
Reduction soybean protein isolate is dissolved in the viscosity after water, and effectively reduces the pimple quantity formed in course of dissolution, goes forward side by side
One step reduces the content of amino-acid nitrogen, makes with amino acid nitrogen content preferably, and effectively increase soybean separation protein
White dispersiveness.
Test example 2
Microculture experiment, test method are carried out to soybean protein isolate prepared by embodiment 1-5 and comparative example 1-3
It is as follows：
The 5g embodiments 1-5 and comparative example 1-3 soybean protein isolates prepared are dissolved in 100mL basal mediums YPD trainings
Support in base, while setting a blank cultures to be 100mL basal medium YPD culture mediums, by S. cervisiae (CICC1921)
It is inoculated into respectively in above-mentioned culture medium with 1% inoculum concentration, 30 DEG C of shaking tables, 150 revs/min of shaken cultivation 30h, tests clump count.
In above-mentioned basal medium YPD culture mediums (yeast extract powder peptone dextrose culture-medium)：Dusty yeast 1%, peptone
2%, glucose 2%, remaining is water, and it is 6.0,121 DEG C of high pressure steam sterilization 20min to adjust pH.
Test result is as follows：
Comprehensive analysis, soybean protein isolate prepared by the present invention is added after culture medium, and microorganism growth situation is good, micro- life
Thing product yield high, illustrates that the soybean protein isolate for preparing of the present invention is beneficial to digesting and assimilating and growing utilization for microorganism.
Claims (10)
1. a kind of preparation method of microorganism specialized soybean protein isolate, including step are as follows：
(1) alkali soluble：At a temperature of 50-55 DEG C, defatted soybean meal is dipped in the aqueous slkali that pH value is 9.5~10.0, adjusts pH
It is worth for 6.8~7.4, pol is 8.0~9.0, stirs 40-60min；Through separation of solid and liquid, protein crude extract is obtained；
(2) acid is heavy：At room temperature, acid solution is added in the protein crude extract obtained to step (1), 10-50min is stirred, through centrifugation
Obtain the heavy albumen of acid；
(3) enzymolysis is neutralized：At room temperature, the heavy albumen of acid step (2) obtained is soluble in water, adjusts pH to 6.00~6.30, pol
For 18.0~19.5, sour heavy protein liquid is obtained；Neutral proteinase and alkali protease are added, at 50-60 DEG C, 0.5-3h is reacted,
Obtain enzymolysis liquid；
(4) refine：The enzymolysis liquid that step (3) is obtained produces microorganism specialized soybean through sterilization, flash distillation process, spray drying
Protein isolate.
2. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(1) mass ratio of defatted soybean meal described in and aqueous slkali is 1:7-11；
It is preferred that, the mass ratio of defatted soybean meal and aqueous slkali described in step (1) is 1:9.5.
3. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(1) aqueous slkali described in is the NaOH or KOH aqueous solution；The NaOH or KOH for being 20-40% with mass concentration in step (3)
The aqueous solution adjusts pH.
4. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(1) pH is adjusted using acid solution in, the mass concentration of the acid solution is 30-35%；
It is preferred that, the acid solution is the aqueous solution of hydrochloric acid, sulfuric acid, phosphoric acid or glacial acetic acid.
5. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(2) acid solution described in is the aqueous hydrochloric acid solution that mass concentration is 20-40%, the pH of regulatory protein crude extract for 4.50~
4.6；The heavy albumen curd dehydration 50~56% of acid described in step (2).
6. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(3) mass concentration of the heavy albumen of acid is 40-50% described in the heavy protein liquid of acid.
7. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(3) mass ratio of acid described in heavy albumen, neutral proteinase and alkali protease is 100：0.8~1.2：1.4~1.8.
8. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(4) method for disinfection is described in：It it is 145~165 DEG C in sterilization temperature, vacuum is progress under the conditions of -0.005~0.03Mpa
Sterilize twice, sterilizing time is respectively 10-30s and 40-90s, the sterilization temperature twice and vacuum all same.
9. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(4) flash conditions are described in：Flash distillation liquid level is 45-55%, and flash vacuum degree is -0.005~-0.015Mpa.
10. the preparation method of microorganism specialized soybean protein isolate according to claim 1, it is characterised in that step
(4) spray drying condition is described in：Wind pushing temperature is 190~220 DEG C, and air inducing temperature is 80~90 DEG C, using liquid phosphatide
Sprayed, the spraying ratio of liquid phosphatide is 4 ‰.
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| CN109730190A (en) * | 2019-02-22 | 2019-05-10 | 黑龙江冰泉多多保健食品有限责任公司 | A kind of high emulsibility soybean protein powder producing method |
| CN110229852A (en) * | 2019-06-20 | 2019-09-13 | 东莞理工学院 | A method of utilizing soybean protein hydrolysate fermenting and producing gamma-polyglutamic acid |
| CN111011579A (en) * | 2019-12-27 | 2020-04-17 | 武汉中地西能科技有限公司 | Black bean selenium polypeptide and preparation method and application thereof |
| CN112273527A (en) * | 2020-08-23 | 2021-01-29 | 山东嘉华保健品股份有限公司 | Production process of soybean protein isolate with extremely low viscosity and excellent flavor |
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| CN107821731A (en) * | 2017-11-07 | 2018-03-23 | 山东禹王生态食业有限公司 | A kind of Semen sojae atricolor enzymatic protein spray coating liquor and preparation method thereof |
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| CN109730190A (en) * | 2019-02-22 | 2019-05-10 | 黑龙江冰泉多多保健食品有限责任公司 | A kind of high emulsibility soybean protein powder producing method |
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| CN110229852B (en) * | 2019-06-20 | 2021-02-23 | 东莞理工学院 | Method for producing gamma-polyglutamic acid by fermenting soybean protein zymolyte |
| CN111011579A (en) * | 2019-12-27 | 2020-04-17 | 武汉中地西能科技有限公司 | Black bean selenium polypeptide and preparation method and application thereof |
| CN112273527A (en) * | 2020-08-23 | 2021-01-29 | 山东嘉华保健品股份有限公司 | Production process of soybean protein isolate with extremely low viscosity and excellent flavor |
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