CN107365819A - A kind of corn sobering-up peptide and its production and use - Google Patents
A kind of corn sobering-up peptide and its production and use Download PDFInfo
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- CN107365819A CN107365819A CN201710711111.7A CN201710711111A CN107365819A CN 107365819 A CN107365819 A CN 107365819A CN 201710711111 A CN201710711111 A CN 201710711111A CN 107365819 A CN107365819 A CN 107365819A
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- corn
- peptide
- enzyme
- sobering
- protein powder
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- 240000008042 Zea mays Species 0.000 title claims abstract description 120
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 title claims abstract description 120
- 235000002017 Zea mays subsp mays Nutrition 0.000 title claims abstract description 120
- 235000005822 corn Nutrition 0.000 title claims abstract description 120
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 60
- 238000004519 manufacturing process Methods 0.000 title abstract description 10
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 79
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 79
- 102000004190 Enzymes Human genes 0.000 claims abstract description 68
- 108090000790 Enzymes Proteins 0.000 claims abstract description 68
- 239000000843 powder Substances 0.000 claims abstract description 61
- 239000007788 liquid Substances 0.000 claims abstract description 33
- 108091005804 Peptidases Proteins 0.000 claims abstract description 29
- 239000004365 Protease Substances 0.000 claims abstract description 27
- 230000000694 effects Effects 0.000 claims abstract description 27
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 27
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims abstract description 25
- 239000006228 supernatant Substances 0.000 claims abstract description 21
- 240000006439 Aspergillus oryzae Species 0.000 claims abstract description 20
- 235000002247 Aspergillus oryzae Nutrition 0.000 claims abstract description 20
- 229920002472 Starch Polymers 0.000 claims abstract description 12
- 239000003513 alkali Substances 0.000 claims abstract description 12
- 235000019698 starch Nutrition 0.000 claims abstract description 12
- 239000008107 starch Substances 0.000 claims abstract description 12
- 238000002360 preparation method Methods 0.000 claims abstract description 8
- 238000005119 centrifugation Methods 0.000 claims abstract description 7
- 102000035195 Peptidases Human genes 0.000 claims abstract description 4
- 239000008367 deionised water Substances 0.000 claims description 21
- 229910021641 deionized water Inorganic materials 0.000 claims description 21
- 238000000034 method Methods 0.000 claims description 13
- 238000003756 stirring Methods 0.000 claims description 10
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 230000009849 deactivation Effects 0.000 claims description 7
- 238000001556 precipitation Methods 0.000 claims description 7
- 238000010008 shearing Methods 0.000 claims description 6
- 238000001816 cooling Methods 0.000 claims description 5
- 238000001035 drying Methods 0.000 claims description 5
- 235000013305 food Nutrition 0.000 claims description 5
- 239000000047 product Substances 0.000 claims description 5
- 239000003814 drug Substances 0.000 claims description 4
- 230000036541 health Effects 0.000 claims description 4
- 238000000605 extraction Methods 0.000 claims description 3
- 239000002054 inoculum Substances 0.000 claims description 2
- 101710093543 Probable non-specific lipid-transfer protein Proteins 0.000 claims 1
- 235000018102 proteins Nutrition 0.000 abstract description 76
- 235000019419 proteases Nutrition 0.000 abstract description 23
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 abstract description 8
- 230000007062 hydrolysis Effects 0.000 abstract description 7
- 238000006460 hydrolysis reaction Methods 0.000 abstract description 7
- 230000006698 induction Effects 0.000 abstract description 6
- 108010068370 Glutens Proteins 0.000 abstract description 5
- 239000004382 Amylase Substances 0.000 abstract description 4
- 102000013142 Amylases Human genes 0.000 abstract description 4
- 108010065511 Amylases Proteins 0.000 abstract description 4
- 235000019418 amylase Nutrition 0.000 abstract description 4
- 235000021312 gluten Nutrition 0.000 abstract description 4
- 229910052757 nitrogen Inorganic materials 0.000 abstract description 4
- 230000000050 nutritive effect Effects 0.000 abstract description 4
- 239000001963 growth medium Substances 0.000 abstract description 3
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- 239000000284 extract Substances 0.000 abstract description 2
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- 238000011084 recovery Methods 0.000 description 27
- 108010021809 Alcohol dehydrogenase Proteins 0.000 description 22
- 102000007698 Alcohol dehydrogenase Human genes 0.000 description 15
- 239000007787 solid Substances 0.000 description 12
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 11
- -1 hydroxyl radical free radical Chemical class 0.000 description 11
- 238000002156 mixing Methods 0.000 description 8
- 239000000523 sample Substances 0.000 description 7
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- 101710176384 Peptide 1 Proteins 0.000 description 4
- 230000000052 comparative effect Effects 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 230000002401 inhibitory effect Effects 0.000 description 4
- 210000004185 liver Anatomy 0.000 description 4
- 229920002494 Zein Polymers 0.000 description 3
- 150000001413 amino acids Chemical class 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 239000002994 raw material Substances 0.000 description 3
- 230000028327 secretion Effects 0.000 description 3
- 239000005019 zein Substances 0.000 description 3
- 229940093612 zein Drugs 0.000 description 3
- DGEZNRSVGBDHLK-UHFFFAOYSA-N [1,10]phenanthroline Chemical compound C1=CN=C2C3=NC=CC=C3C=CC2=C1 DGEZNRSVGBDHLK-UHFFFAOYSA-N 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 238000007796 conventional method Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 230000031700 light absorption Effects 0.000 description 2
- 230000004060 metabolic process Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 239000012488 sample solution Substances 0.000 description 2
- 239000013049 sediment Substances 0.000 description 2
- 239000000243 solution Substances 0.000 description 2
- 238000004659 sterilization and disinfection Methods 0.000 description 2
- QCVGEOXPDFCNHA-UHFFFAOYSA-N 5,5-dimethyl-2,4-dioxo-1,3-oxazolidine-3-carboxamide Chemical compound CC1(C)OC(=O)N(C(N)=O)C1=O QCVGEOXPDFCNHA-UHFFFAOYSA-N 0.000 description 1
- 208000007848 Alcoholism Diseases 0.000 description 1
- 102000009091 Amyloidogenic Proteins Human genes 0.000 description 1
- 108010048112 Amyloidogenic Proteins Proteins 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 108010016626 Dipeptides Proteins 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000006395 Globulins Human genes 0.000 description 1
- 108010044091 Globulins Proteins 0.000 description 1
- 208000001953 Hypotension Diseases 0.000 description 1
- QIVBCDIJIAJPQS-VIFPVBQESA-N L-tryptophane Chemical compound C1=CC=C2C(C[C@H](N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-VIFPVBQESA-N 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 description 1
- 239000004472 Lysine Substances 0.000 description 1
- 240000007594 Oryza sativa Species 0.000 description 1
- 235000007164 Oryza sativa Nutrition 0.000 description 1
- YNPNZTXNASCQKK-UHFFFAOYSA-N Phenanthrene Natural products C1=CC=C2C3=CC=CC=C3C=CC2=C1 YNPNZTXNASCQKK-UHFFFAOYSA-N 0.000 description 1
- QIVBCDIJIAJPQS-UHFFFAOYSA-N Tryptophan Natural products C1=CC=C2C(CC(N)C(O)=O)=CNC2=C1 QIVBCDIJIAJPQS-UHFFFAOYSA-N 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- IKHGUXGNUITLKF-XPULMUKRSA-N acetaldehyde Chemical compound [14CH]([14CH3])=O IKHGUXGNUITLKF-XPULMUKRSA-N 0.000 description 1
- 230000003213 activating effect Effects 0.000 description 1
- 201000007930 alcohol dependence Diseases 0.000 description 1
- 230000001476 alcoholic effect Effects 0.000 description 1
- 230000002075 anti-alcohol Effects 0.000 description 1
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- 239000000427 antigen Substances 0.000 description 1
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- 108091007433 antigens Proteins 0.000 description 1
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- 239000007864 aqueous solution Substances 0.000 description 1
- 239000002585 base Substances 0.000 description 1
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- 239000008280 blood Substances 0.000 description 1
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- 239000006172 buffering agent Substances 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 239000005515 coenzyme Substances 0.000 description 1
- 238000010411 cooking Methods 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 239000000287 crude extract Substances 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 235000014103 egg white Nutrition 0.000 description 1
- 210000000969 egg white Anatomy 0.000 description 1
- 239000000686 essence Substances 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 239000000413 hydrolysate Substances 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 description 1
- 208000021822 hypotensive Diseases 0.000 description 1
- 230000001077 hypotensive effect Effects 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229920002521 macromolecule Polymers 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 229930014626 natural product Natural products 0.000 description 1
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- 230000003647 oxidation Effects 0.000 description 1
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- 239000008363 phosphate buffer Substances 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 238000002203 pretreatment Methods 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
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- 238000012827 research and development Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 108091008146 restriction endonucleases Proteins 0.000 description 1
- 235000009566 rice Nutrition 0.000 description 1
- 230000001568 sexual effect Effects 0.000 description 1
- 238000004904 shortening Methods 0.000 description 1
- 238000010025 steaming Methods 0.000 description 1
- 238000010189 synthetic method Methods 0.000 description 1
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- 231100000419 toxicity Toxicity 0.000 description 1
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Classifications
-
- 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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/346—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of vegetable proteins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/01—Hydrolysed proteins; Derivatives thereof
- A61K38/011—Hydrolysed proteins; Derivatives thereof from plants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/14—Fungi; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The invention discloses a kind of corn sobering-up peptide and its production and use, the preparation method comprises the following steps：Corn protein powder is taken, adds water and starch, is then inoculated with aspergillus oryzae, be put into incubator and cultivate；2 5min are sheared after taking-up, 1 2h is stirred at 30 40 DEG C and extracts thick enzyme；Corn protein powder is taken, adds water, alkali protease and thick enzyme, is adjusted to required pH value, after digesting 4 6h, centrifugation, supernatant is corn sobering-up peptide liquid.The present invention uses culture media nitrogen source induction aspergillus oryzae extracellular proteinase based on corn protein powder, and addition starch induction aspergillus oryzae enzyme secreting amylase, producing thick enzyme has higher protease activity and starch hydrolysis activity, can efficient hydrolysised corn gluten.The self-control enzyme of the present invention is safe, in addition to energy effectively hydrolyzing corn protein powder improves nutritive value, moreover it is possible to removes the factor for causing corn sobering-up peptide to produce bitter taste.
Description
Technical field
The present invention relates to a kind of corn sobering-up peptide and its production and use.
Background technology
In the cooking culture in China, there is dinner to have wine.But the World Health Organization proposes in portion is reported：In alcohol
Poison is one big public hazards of the world today, and its toxicity can be accumulated in whole body main organs, causes alcoholic liver injury, or it is some other simultaneously
Send out disease.Progress at present using synthetic method development antialcoholic drugs is difficult, and the research on natural products sobering-up functions is more and more
Attract attention.
Corn is one of Three major grain crops, is the important raw material of industry again.Corn protein powder is production of corn starch
Principal by product, about containing 60% protein, mainly contain alcohol soluble protein (68%), glutelin (22%), globulin
(1.2%).Due to zein coarse mouthfeel, poorly water-soluble, lack the essential amino acid such as lysine, tryptophan, nutritive value
It is not high, also with peculiar smell, its application in field of food is limited, is sold frequently as low value feed, so to corn protein powder
Carry out deep processing or improve its nutritive value using the peptide based food of the hydrolysis production high nutrition of protease, for lengthening manufacturing chain
Bar, raising economic value added have certain meaning.
Polypeptide is that natural amino acid is formed with different form with arrangement mode in protein, from dipeptides to the linear of complexity
Or the general name of the different peptides of loop configuration.Polypeptide absorption easier than protein, low antigen, biological value specific ionization amino acid will
It is high.In addition, polypeptide also has a variety of physiologically actives, such as hypotensive, anti-oxidant, antifatigue.
By the effect of protease, corn protein powder can be degraded to the different corn peptide of peptide chain length.Corn peptide is not only
With good solubility, also with a variety of physiologically actives, wherein relieving alcoholism and protecting liver is one of its important activity.In recent years, wake up
The research and development of wine product are in the ascendant, and especially protein-based production development of sobering up is even more to be a dark horse, and corn sobering-up peptide is wherein
Outstanding person.Corn peptide can promote alcohol metabolism, reduce ethanol in blood and its content of oxide acetaldehyde, can shorten drunk person
Sober up the time, while also there is liver protection function.Result of study shows that corn peptide is by suppressing hydroxyl free free radical, swashing
The metabolism of alcohol dehydrogenase accelerating alcohol living, so as to mitigate damage of the alcohol in high concentration to liver.
Native corn protein macromolecule water-solubility is poor, stereochemical structure is close, and property is extremely stable.Remained in corn protein powder
There is a certain amount of starch, the restriction enzyme site of protein in corn protein powder is tightly wrapped up, so as to the serious shadow of factor
The hydrolysis rate of zein and the utilization rate of raw material have been rung, has caused corn peptide yield relatively low.So hydrolyzed in corn protein powder
Before be necessary carry out pre-treatment, improve its hydrolysis efficiency.
The content of the invention
In order to solve the problems, such as to cause hydrolysis efficiency and protein recovery low due to starch in corn protein powder, the present invention
Primary and foremost purpose be to provide a kind of preparation method of corn sobering-up peptide, culture of this method using corn protein powder as aspergillus oryzae
Base nitrogen source, the secretion of induction aspergillus oryzae can effectively hydrolyzing corn protein powder protease and amylase, by this obtained protease and
Amylase crude extract combination commercial enzyme cohydrolysis corn protein powder, corn sobering-up peptide is made.
Another object of the present invention is to provide the corn sobering-up peptide as made from the above method.
It is still another object of the present invention to provide the purposes of above-mentioned corn sobering-up peptide.
The purpose of the present invention is achieved through the following technical solutions：
A kind of preparation method of corn sobering-up peptide, comprises the following steps：
(1) corn protein powder is weighed, the deionized water of 0.8-2.0 times of weight is added, sterilizes after stirring evenly, add at room temperature
Corn protein powder weight 20-30% starch (as carbon source), aspergillus oryzae is inoculated with, is put into incubator and cultivates after stirring evenly；Take out
After add deionized water, make solid-liquid ratio (weight ratio) be 1:8-1:15, after 5-7kr/min speed shearing 2-5min, in 30-
1-2h extraction protease is stirred at 40 DEG C, (8000-12000) × g centrifuges 20-30min at 0-4 DEG C, collects supernatant, and concentration is cold
It is lyophilized it is dry after produce thick enzyme；
Sterilizing described in step (1) is the 15-30min that sterilized at 100-121 DEG C；
Make 3042 in the preferred Shanghai of aspergillus oryzae described in step (1)；The inoculum concentration of the aspergillus oryzae is (10-30) × 109Individual spore
Son/gram dry medium；
The condition of culture and cultural method of step (1) the incubator culture be：30-35 DEG C of temperature, humidity 90-98%,
12-16h is cultivated, turns over Qu Yici, once song is turned over again every 6h, adjusts 25-30 DEG C of temperature, humidity 85-90%, be further cultured for 24-30h and take
Go out；
(2) corn protein powder of 60-100 mesh sieves was taken to add deionized water, it is 1 to make solid-liquid ratio:8-1:15, add alkalescence
Thick enzyme made from protease and step (1), alkali protease and thick enzyme respectively account for the 0.3-0.8% of corn protein powder weight, are adjusted to
Required pH value (according to the difference of the alkali protease specifically used, required pH value is different), 4-6h is digested at 45-60 DEG C of temperature
Afterwards, enzyme deactivation cools down, and centrifuges 15-30min in (5000-8000) × g, removes precipitation, supernatant is corn sobering-up peptide liquid；
37071 alkali proteases of the preferred Novozymes Company of alkali protease described in step (2), enzymolysis pH value are 7.5-
8.5。
The corn sobering-up peptide as made from the above method has higher hydroxyl radical free radical clearance rate and alcohol dehydrogenase enzyme activition
Rate, available for medicine, health products and food of the preparation with effect of sobering up.
The present invention is had the following advantages relative to prior art and effect：
(1) present invention uses culture media nitrogen source induction aspergillus oryzae extracellular proteinase based on corn protein powder, and addition is formed sediment
Powder induces aspergillus oryzae enzyme secreting amylase, and producing thick enzyme has higher protease activity and starch hydrolysis activity, can be efficient
Hydrolysised corn gluten.
(2) self-control enzyme of the invention is safe, in addition to energy effectively hydrolyzing corn protein powder improves nutritive value, moreover it is possible to go
Except the factor for causing corn sobering-up peptide to produce bitter taste, and the generation of anti-nutritional factors can be reduced, thus addition can be reduced
The necessity of bitter equipment is taken off, production technology simplifies, and has effectively saved cost.
(3) present invention process is simple to operate, production cost is low, high without any pollution, protein recovery, gained antialcoholism peptide
Hydroxyl radical free radical clearance rate and alcohol dehydrogenase activity ratio it is high, bioactivity is high, can be widely applied to medicine, health products and food
In product field.
Embodiment
With reference to embodiment, the present invention is described in further detail, but the implementation of the present invention is not limited to this.
In the present invention, alcohol dehydrogenase activity ratio is determined as follows：
Sample cell：0.65ml buffering agents one (1 are sequentially added into test tube:1 ultra-pure water dilutes), 0.05ml reagents two
The existing liquid of ethanol, 0.03ml ADH and 0.02ml sample solutions.After fully mixing, take and determine the μ L of mixed liquor 100 and be placed in clean ultraviolet enzyme
In target hole, 37 DEG C of incubation 5min, ejection adds 100 μ L coenzyme reagents three (10mL ultra-pure waters dilute before use), pushes away immediately
Enter and it is whole keep constant temperature, every 10s sweep measurings, once light absorption value ends to 10min in each reacting hole under 340nm, line taking
Initial value is A1 in sexual intercourse sound response curve, and end value is that (reagent one, reagent two, reagent three come from alcohol dehydrogenase to A2
Kit).
Control tube：Sample, the same sample cell of remaining step are replaced with deionized water.
ADH activity ratio formula are as follows：
In the present invention, the measure of hydroxyl radical free radical clearance rate is as follows using o-phenanthroline：
Reaction is with H2O2/Fe2+System produces OH, net reaction by Fenton reactions：
Fe2++H2O2→Fe3++·OH+OH-
Fe(C12H8N2)3 2+(orange red)+OH → Fe (C12H8N2)3 3+(light blue)
Phen-Fe2+The aqueous solution is oxidized to Phen-Fe by hydroxyl radical free radical3+Afterwards, it inhales in 536nm maximum
Receive peak to disappear, according to principles above, with A536Change reflection OH oxidation.
Specifically determining operation is：
Sample cell：Take 0.6ml Phen solution (5mmolL-1) add 0.4ml phosphate buffer (0.2M, pH value
7.4) after mixing, 0.6ml sample solutions and 0.6ml EDTA (15mmolL are added-1), add 0.6ml after mixing again
FeSO4(5mmol·L-1) and deionized water mend to volume 2.8ml, add 0.8ml H after fully mixing2O2(0.1%), shake up
1h is incubated after 37 DEG C, surveys light absorption value at 536nm.
Damage pipe：Sample, the same sample cell of remaining step are replaced with deionized water.
Pipe is not damaged：Deionized water replaces H2O2, remaining step is same to damage pipe.
OH clearance rates (%)=(A536 sample cells-A536 damage pipes)/(A536 do not damage pipe-A536Damage pipe)×100
The measure of alcohol dehydrogenase activity ratio uses alcohol dehydrogenase enzyme reagent kit.
Embodiment 1
A kind of preparation method of corn sobering-up peptide, comprises the following steps：
(1) fermentation prepares thick enzyme：Corn protein powder is weighed, 0.8 times of deionized water is added, stirs, be put into high pressure
100 DEG C of sterilizing 30min, are cooled to room temperature in steam sterilization pan after taking-up, add 30% starch of corn protein powder weight, connect
Kind aspergillus oryzae (making 3042 in Shanghai) (10 × 109Individual spore/gram dry medium), it is put into after mixing uniformly in incubator, adjusts the temperature of incubator
30 DEG C of degree, humidity 90%, 15h is cultivated, turns over Qu Yici, turned over once song again every 6h, adjust 28 DEG C of temperature, humidity 85%, be further cultured for
27h, which takes out, adds deionized water, and it is 1 to make solid-liquid ratio:8, after 5kr/min speed shearing 3min, stirring 1h is carried at 40 DEG C
Take protease, 12000 × g centrifugations 20min at 0 DEG C, collect supernatant, concentrated frozen produces thick enzyme after drying.
(2) thick enzyme and commercial enzyme cohydrolysis corn protein powder are made by oneself：Took 80 mesh sieves corn protein powder add go from
Sub- water, it is 1 to make solid-liquid ratio:8, add and make thick enzyme and the alkali protease of Novozymes Company 37071 by oneself, enzyme bottom is adjusted than being respectively 0.8%
PH value 8.5 is saved, after digesting 6h under the conditions of temperature 45 C, enzymolysis liquid is in 95 DEG C of enzyme deactivation 15min, and 8000 × g is centrifuged after cooling
15min, precipitation is removed, supernatant is corn sobering-up peptide liquid.
(3) gained supernatant is spray-dried obtains corn sobering-up peptide 1.
Solid recovery rate, protein recovery, hydroxyl radical free radical clearance rate and the alcohol dehydrogenase activity ratio knot of enzymolysis liquid
Fruit is shown in Table 1~2 respectively.
Embodiment 2
A kind of preparation method of corn sobering-up peptide, comprises the following steps：
(1) fermentation prepares thick enzyme：Corn protein powder is weighed, 1.5 times of deionized water is added, stirs, be put into high pressure
121 DEG C of sterilizing 15min in steam sterilization pan, room temperature, the starch of addition corn protein powder weight 20%, inoculation are cooled to after taking-up
Aspergillus oryzae (making 3042 in Shanghai) (30 × 109Individual spore/gram dry medium), it is put into after mixing uniformly in incubator, adjusts the temperature of incubator
32 DEG C, humidity 98%, 12h is cultivated, turns over Qu Yici, turned over once song again every 6h, adjust 30 DEG C of temperature, humidity 90%, be further cultured for 24h
Take out and add deionized water, it is 1 to make solid-liquid ratio:15, after 6.5kr/min speed shearing 2min, stirring 2h is carried at 30 DEG C
Take protease, 8000 × g centrifugations 30min at 4 DEG C, collect supernatant, concentrated frozen produces thick enzyme after drying.
(2) thick enzyme and commercial enzyme cohydrolysis corn protein powder are made by oneself：Took 60 mesh sieves corn protein powder add go from
Sub- water, it is 1 to make solid-liquid ratio:15, add and make thick enzyme and the alkali protease of Novozymes Company 37071 by oneself, enzyme bottom ratio is respectively 0.5%,
PH value 8.0 is adjusted, after digesting 4h under the conditions of temperature 60 C, enzymolysis liquid is in 95 DEG C of enzyme deactivation 15min, and 5000 × g is centrifuged after cooling
30min, precipitation is removed, supernatant is corn sobering-up peptide liquid.
(3) gained supernatant is spray-dried obtains corn sobering-up peptide 2.
Solid recovery rate, protein recovery, hydroxyl radical free radical clearance rate and the alcohol dehydrogenase activity ratio knot of enzymolysis liquid
Fruit is shown in Table 1~2 respectively.
Embodiment 3
A kind of preparation method of corn sobering-up peptide, comprises the following steps：
(1) fermentation prepares thick enzyme：Corn protein powder is weighed, 2 times of deionized water is added, stirs, is put into high pressure steaming
115 DEG C of sterilizing 20min in vapour autoclave, room temperature is cooled to after taking-up, the starch of addition corn protein powder weight 25%, is inoculated with rice
Aspergillus (making 3042 in Shanghai) (20 × 109Individual spore/gram dry medium), it is put into after mixing uniformly in incubator, adjusts the temperature 35 of incubator
DEG C, humidity 95%, 16h is cultivated, turns over Qu Yici, turned over once song again every 6h, adjust 25 DEG C of temperature, humidity 88%, be further cultured for 30h and take
Go out to add deionized water, it is 1 to make solid-liquid ratio:12, after 7kr/min speed shearing 5min, 1.5h extractions are stirred at 35 DEG C
Protease, 10000 × g centrifugations 25min at 2 DEG C, collects supernatant, and concentrated frozen produces thick enzyme after drying.
(2) thick enzyme and commercial enzyme cohydrolysis corn protein powder are made by oneself：Took 100 mesh sieves corn protein powder add go from
Sub- water, it is 1 to make solid-liquid ratio:12, add and make thick enzyme and the alkali protease of Novozymes Company 37071 by oneself, enzyme bottom ratio is respectively 0.3%,
PH value 7.5 is adjusted, after digesting 5h under the conditions of 55 DEG C of temperature, enzymolysis liquid is in 95 DEG C of enzyme deactivation 15min, and 6000 × g is centrifuged after cooling
20min, precipitation is removed, supernatant is corn sobering-up peptide liquid.
(3) gained supernatant is spray-dried obtains corn sobering-up peptide 3.
Solid recovery rate, protein recovery, hydroxyl radical free radical clearance rate and the alcohol dehydrogenase activity ratio knot of enzymolysis liquid
Fruit is shown in Table 1~2 respectively.
Comparative example 1
Antialcoholism peptide is prepared using conventional method (non-invention method) water extraction corn protein powder：
(1) water extraction of corn protein powder：Corn protein powder added deionized water after taking 80 mesh sieves, and it is 1 to make solid-liquid ratio:
15, it is put into the shaking table of 55 DEG C of temperature and shakes 5h, take out the 15min that sterilized at 95 DEG C, is cooled in 8000 × g centrifugation 15min, it is heavy to remove
Form sediment, supernatant is the peptide mixer of sobering up of corn protein powder.
(2) gained supernatant is spray-dried obtains corn peptide 1.
Solid recovery rate, protein recovery, hydroxyl radical free radical clearance rate and the alcohol dehydrogenase activity ratio knot of enzymolysis liquid
Fruit is shown in Table 1~2 respectively.
Comparative example 2
Using the thick enzyme hydrolyzed corn albumen powder of self-control：
(1) fermentation prepares thick enzyme：Raw material corn protein powder is weighed, 0.8 times of deionized water is added, stirs, be put into
121 DEG C of sterilizing 15min, are cooled to room temperature in high-pressure steam sterilizing pan after taking-up, add the starch of corn protein powder weight 30%,
It is inoculated with aspergillus oryzae (making 3042 in Shanghai) (20 × 109Individual spore/gram dry medium), it is put into after mixing uniformly in incubator, adjusts incubator
30 DEG C of temperature, humidity 90%, 15h is cultivated, turns over Qu Yici, turn over once bent, 28 DEG C of temperature of regulation, humidity 85% again every 6h, then train
Support 27h and take out addition deionized water, it is 1 to make solid-liquid ratio:15, after 5kr/min speed shearing 5min, 2h is stirred at 40 DEG C
Protease is extracted, 10000 × g centrifuges 30min at 4 DEG C, collects supernatant, and concentrated frozen produces thick enzyme after drying.
(2) thick enzyme hydrolyzed corn albumen powder is made by oneself：Took the corn protein powder of 80 mesh sieves to add deionized water, and made solid-liquid ratio
For 1:15, add the 0.8% thick enzyme of self-control, adjust pH value 8.5, be put under the conditions of 55 DEG C of temperature after digesting 5h, enzymolysis liquid is in 95
DEG C enzyme deactivation 15min, 8000 × g centrifuges 15min after cooling, removes precipitation, supernatant is corn sobering-up peptide liquid.
(3) gained supernatant is spray-dried obtains corn peptide 2.
Solid recovery rate, protein recovery, hydroxyl radical free radical clearance rate and the alcohol dehydrogenase activity ratio knot of enzymolysis liquid
Fruit is shown in Table 1~2 respectively.
Comparative example 3
Antialcoholism peptide is prepared using conventional method (non-invention method) hydrolysised corn gluten：
(1) enzymolysis of corn protein powder：Corn protein powder added deionized water after taking 80 mesh sieves, and it is 1 to make solid-liquid ratio:
15,0.8% alkali protease of Novozymes Company 37071 is added, pH value 8.5 is adjusted, is put into the shaking table of 55 DEG C of temperature and digests
5h, take out in 95 DEG C of enzyme deactivation 15min, be cooled in 8000 × g centrifugation 15min, remove precipitation, supernatant is corn protein powder
Antialcoholism peptide liquid.
(2) gained supernatant is spray-dried obtains corn peptide 3.
Solid recovery rate, protein recovery, hydroxyl radical free radical clearance rate and the alcohol dehydrogenase activity ratio knot of enzymolysis liquid
Fruit is shown in Table 1~2 respectively.
The indices of the corn peptide of table 1
The OH clearance rates and ADH activity ratio results of the corn peptide of table 2
It can be found by table 1, corn protein powder is hydrolyzed jointly using thick enzyme and commercial enzyme is made by oneself, can obtain higher
Solid recovery rate (50.57-52.49%) and protein recovery (50.98-52.86%).Can from corn peptide 1 and corn peptide 2
Go out, using the direct water extraction of corn protein powder (corn peptide 1), the solid recovery rate and protein recovery of water extract are respectively
7.42% and 1.73%, it is very low.And the corn peptide 2 that corn protein powder is obtained with self-control enzyme enzymolysis, its solid recovery rate are
More than 4 times of water extraction, protein recovery are water extractions more than 23 times, and explanation is used as culture media nitrogen source induction aspergillus oryzae by the use of corn protein powder
The protease of secretion is active, and activity is very high.From corn peptide 2 and corn peptide 3 as can be seen that the protein recovery of corn peptide 2
Protein recovery than corn peptide 3 is high, illustrates to make enzyme effect zein amyloid proteins more more preferable than commercial enzyme by oneself, because aspergillus oryzae
The protein in corn protein powder can not directly be utilized, it is necessary to secret out of protease the degradation of proteins into small-molecular peptides or amino
Acid can just absorb, thus the protease of corn protein powder induction aspergillus oryzae secretion have to have corn protein powder it is higher
Hydrolysis efficiency could meet the nutritional need of aspergillus oryzae growth.Exclusive use is can be seen that from corn peptide 2,3 and corn sobering-up peptide
The solid recovery rate and protein recovery of self-control enzyme or commercial enzyme are not used in conjunction with the height of both enzymes.
Table 2 is the hydroxyl radical free radical clearance rate and alcohol dehydrogenase of corn protein powder hydrolysate in each embodiment and comparative example
Enzyme activition rate.Result of study shows, corn sobering-up peptide passes through generation for suppressing hydroxyl radical free radical, activating alcohol dehydrogenase accelerating alcohol
Thank, so as to mitigate damage of the alcohol in high concentration to liver, shortening is sobered up the time.From in table, with self-control enzyme hydrolyzed corn egg
White powder, obtain highest OH inhibiting rates and higher ADH activity ratios.Make enzyme and commercial enzyme cohydrolysis corn protein powder by oneself,
The OH inhibiting rate and ADH activity ratio more slightly lower than self-control enzyme hydrolyzed corn albumen powder are obtained, than commercial enzyme hydrolysised corn gluten
Higher OH inhibiting rates and ADH activity ratios.Show that the art of this patent makes the releasable OH inhibiting rates of enzyme hydrolyzed corn albumen by oneself
The higher corn peptide with ADH activity ratios, but the solid recovery rate and albumen time of self-control enzyme hydrolyzed corn albumen is used alone
Yield is below making enzyme and commercial enzyme collective effect by oneself.
Comprehensive Tables 1 and 2 can be seen that, can be carried using self-control enzyme and commercial enzyme cohydrolysis corn protein powder with highly significant
Solid recovery rate, protein recovery and the absolute sobering-up active of high corn sobering-up peptide, increase factory benefit.
Above-described embodiment is the preferable embodiment of the present invention, but embodiments of the present invention are not by above-described embodiment
Limitation, other any Spirit Essences without departing from the present invention with made under principle change, modification, replacement, combine, simplification,
Equivalent substitute mode is should be, is included within protection scope of the present invention.
Claims (7)
1. a kind of preparation method of corn sobering-up peptide, it is characterised in that comprise the following steps：
(1) corn protein powder is weighed, the deionized water of 0.8-2.0 times of weight is added, is sterilized after stirring evenly, add corn at room temperature
Albumen powder weight 20-30% starch, aspergillus oryzae is inoculated with, is put into incubator and cultivates after stirring evenly；Deionized water is added after taking-up,
It is 1 to make solid-liquid ratio:8-1:15, after 5-7kr/min speed shearing 2-5min, 1-2h extraction albumen is stirred at 30-40 DEG C
Enzyme, (8000-12000) × g centrifugations 20-30min at 0-4 DEG C, collects supernatant, and concentrated frozen produces thick enzyme after drying；
The condition of culture and cultural method of step (1) the incubator culture be：30-35 DEG C of temperature, humidity 90-98%, culture
12-16h, Qu Yici is turned over, once song is turned over again every 6h, adjusted 25-30 DEG C of temperature, humidity 85-90%, be further cultured for 24-30h taking-ups；
(2) corn protein powder of 60-100 mesh sieves was taken to add deionized water, it is 1 to make solid-liquid ratio:8-1:15, add basic protein
Thick enzyme made from enzyme and step (1), alkali protease and thick enzyme respectively account for the 0.3-0.8% of corn protein powder weight, are adjusted to required
PH value, after digesting 4-6h at 45-60 DEG C of temperature, enzyme deactivation cooling, 15-30min is centrifuged in (5000-8000) × g, removes precipitation,
Supernatant is corn sobering-up peptide liquid.
2. according to the method for claim 1, it is characterised in that：Sterilizing described in step (1) is sterilized at 100-121 DEG C
15-30min。
3. according to the method for claim 1, it is characterised in that：Aspergillus oryzae described in step (1) makes 3042 for Shanghai.
4. according to the method for claim 1, it is characterised in that：The inoculum concentration of step (1) described aspergillus oryzae be (10-30) ×
109Individual spore/gram dry medium.
5. according to the method for claim 1, it is characterised in that：Alkali protease described in step (2) is Novozymes Company
37071 alkali proteases, enzymolysis pH value is 7.5-8.5.
A kind of 6. corn sobering-up peptide, it is characterised in that：It is to be made as the method described in claim any one of 1-5.
7. the corn sobering-up peptide described in claim 6 is being prepared with the application in medicine, health products or the food for sobering up effect.
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| CN110747247A (en) * | 2018-07-23 | 2020-02-04 | 江苏挺卫实业有限公司 | Separation and extraction method of corn peptide for anti-alcohol beverage |
| CN114469773A (en) * | 2022-02-16 | 2022-05-13 | 广东迈高生物科技研究有限公司 | Stable anti-aging composition and preparation and application thereof |
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