CN107841518A - A kind of krill anti-oxidation peptide and preparation method and application - Google Patents
A kind of krill anti-oxidation peptide and preparation method and application Download PDFInfo
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- CN107841518A CN107841518A CN201711275246.XA CN201711275246A CN107841518A CN 107841518 A CN107841518 A CN 107841518A CN 201711275246 A CN201711275246 A CN 201711275246A CN 107841518 A CN107841518 A CN 107841518A
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- krill
- oxidation peptide
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- seed liquor
- bacillus
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- 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
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- 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
Abstract
The invention belongs to the technical field of biological fermentation engineering, discloses a kind of krill anti-oxidation peptide and preparation method and application.Methods described is：(1) degreasing euphausia superba powder, water and carbon source are mixed, regulation pH value of solution is 6.5~7.5, sterilizing, obtains euphausia superba powder solution；(2) bacillus subtilis and bacillus coagulans are inoculated in culture medium respectively, shaking table culture, adjust the concentration of spore, obtain bacillus subtilis seed liquor and bacillus coagulans seed liquor；(3) above two seed liquor is compounded, obtains compounding bacterium solution；Compounding bacterium solution is mixed with euphausia superba powder solution again, shaker fermentation, centrifugation, supernatant concentration, dries, obtains krill anti-oxidation peptide.The method of the present invention is simple；Anti-oxidation peptide rich in probiotics, has preferable antioxidation activity, the comprehensive utilization ratio of krill protein is improved, for health products and/or field of functional food without shrimp fishy smell.
Description
Technical field
The invention belongs to biological fermentation engineering field, is related to a kind of with bacillus subtilis and the compounding of bacillus coagulans
A kind of method that strain fermentation processing krill obtains the anti-oxidation peptide rich in probiotics, and in particular to krill is anti-oxidant
Peptide and preparation method and application.
Background technology
Krill (Euphausia Superba) is one of biology that biomass is maximum on the earth, procreation is most successful,
It is a kind of important ocean new resources.Exploitation of the China to krill is still in the junior stage, and is mainly reflected in krill oil
Or in the extraction of astaxanthin, the recycling for krill albumen is relatively fewer.At present, krill crude protein is used as low value more
Fish meal, and use strong acid and strong base method during extraction more, yield is low, and protein structure is destroyed, and nutritive value is low；And by krill egg
White obtained krill peptide has multiple biological activities, such as anti-oxidant, antibacterial, antifatigue etc., is led in functional food and cosmetics
Domain possesses huge application potential.But krill peptide is usually associated with dense heavy shrimp fishy smell in preparation process, limits it
Follow-up development and application.Therefore, find suitable process krill albumen is processed and deodorization be improve krill money
The key of source comprehensive utilization.
Bacillus subtilis is the safe strain that Food and Drug Administration announces, and is using most in current industrial production
One of extensive strain.By suitable fermentation condition, bacillus subtilis can produce the higher neutral proteinase of vigor, energy
Enough it is applied among food industry.Bacillus coagulans (Bacillus coagulans) is rear of field of probiotic bacteria in recent years
Show, in addition to the health-care efficacy with lactic acid bacteria and Bifidobacterium etc., also there is abundant enzyme system and high temperature resistant, acidproof
With bile tolerance grade height anti-adversity, and there is stronger suppression pathogenic entero becteria ability.Have been widely used for abroad medicine,
The industries such as food, health care, herding and aquatic products.
The present invention is fermented using the strain after bacillus subtilis and bacillus coagulans compounding to euphausia superba powder,
The shrimp fishy smell of euphausia superba powder is removed, and is with anti-oxidant by krill protein degradation using caused protease in fermentation process
The krill peptide of activity.This measure can not only greatly improve the added value of krill albumen, be provided for health products trade a large amount of high-quality
Raw material, can also be that a new approach is opened up in the deep processing of krill albumen.
The content of the invention
It is an object of the invention to provide a kind of compounding strain fermentation using bacillus subtilis and bacillus coagulans to go
Except the fishy smell of euphausia superba powder, and the method for preparing the krill anti-oxidation peptide rich in probiotics, so as to improve krill albumen
The comprehensive utilization ratio of matter, improve the added value of krill protein product.
Another object of the present invention is to provide the krill anti-oxidation peptide being prepared by the above method.
It is still another object of the present invention to provide the application of above-mentioned krill anti-oxidation peptide.The krill is anti-oxidant
Peptide is applied to health products and/or field of functional food.
To achieve the above object, the present invention adopts the following technical scheme that：
A kind of preparation method of krill anti-oxidation peptide, comprises the following steps：
(1) degreasing euphausia superba powder, water and carbon source are mixed, regulation pH value of solution is 6.5~7.5, sterilizing, obtains South Pole phosphorus
Shrimp med solution；
(2) bacillus subtilis and bacillus coagulans are inoculated in fluid nutrient medium respectively, shaken under the conditions of 30~45 DEG C
Bed 20~24h of culture, adjusts the concentration of spore, obtains bacillus subtilis seed liquor and bacillus coagulans seed liquor；
(3) by bacillus subtilis seed liquor and bacillus coagulans seed liquor according to volume ratio 1:1.5~1:4 are answered
Match somebody with somebody, obtain compounding bacterium solution；Compounding bacterium solution is mixed with euphausia superba powder solution again, the shaker fermentation under the conditions of 35~45 DEG C
35~45 hours, centrifugation, by supernatant concentration, dry, obtain krill anti-oxidation peptide.Fermentation time is preferably 35~42 small
When.
The mass ratio of degreasing euphausia superba powder and water described in step (1) is 1:5~1:10；The dosage of the carbon source is de-
The 5%~10% of fat euphausia superba powder and the gross mass of water；Degreasing krill meal be the extracted krill oil of krill meal after it is remaining under
Heel.
Carbon source described in step (1) is more than one in maltose, glucose, lactose, sucrose, soluble starch；
25~45min of moist heat sterilization under the conditions of sterilizing refers to 110~130 DEG C described in step (1), is cooled to after sterilizing
35-45℃；
Fluid nutrient medium obtains by the following method described in step (2)：Peptone 10.0g, beef extract 10.0g are weighed,
Sodium chloride 5.0g, add water to be settled to 1000mL, adjust pH to 6.5~7.5, sterilizing, produce fluid nutrient medium；The sterilizing refers to
25~45min of moist heat sterilization under the conditions of 110~130 DEG C, cools to 35~45 DEG C after sterilizing；
The concentration of bacillus subtilis seed liquor miospore described in step (2) is 106~109Individual/mL；Condense gemma bar
The concentration of bacterium seed liquor miospore is 106~109Individual/mL；
The temperature cultivated described in step (2) is preferably 30~35 DEG C；
The rotating speed of shaking table culture described in step (2) is 100~200rpm；
It is 1 that the spore number of bacillus subtilis and bacillus coagulans ratio in bacterium solution is compounded described in step (3):1.5
~1:4；
The dosage that bacterium solution is compounded described in step (3) is the 5%~10% of euphausia superba powder liquor capacity；
The rotating speed of shaking table culture described in step (3) is 100~150rpm；
Centrifugal rotational speed described in step (3) is that concentration described in 2500~4000r/min is 30~45 DEG C of spin concentrations to 30
~45Bx (soluble solid content 30-45%), the drying are spray drying.
The bacillus subtilis preserving number CICC10075, depositary institution：In the management of Chinese industrial Microbiological Culture Collection
The heart；Bacillus coagulans preserving number CCTCCM2017054, depositary institution：China typical culture collection center.
Fermented using strain is compounded, in the krill peptide of preparation the relative amount of trimethylamine be 2.49%~
5.17%, without obvious shrimp fishy smell, polypeptide molecular weight is 1000-7000Da in product, and content 60-80%, the TEAC of product resist
Oxidability is 60-200 μM of Trolox/mg, and ORAC oxidation resistances are 50-140 μM of Trolox/mg, viable count in product
Amount reaches 1 × 106-107CFUs/g。
The present invention is fermented by compounding bacillus subtilis and bacillus coagulans to degreasing euphausia superba powder, is removed
Shrimp fishy smell, while there is antioxidation activity using protease hydrolytic krill albumen, preparation caused by bacterial strain metabolism in fermentation process
Krill peptide, finally give the krill peptide product rich in bacillus coagulans and B family vitamin.
Euphausia superba powder of the present invention is rich and easy to get, and the above method can remove shrimp fishy smell and prepare rich in prebiotic
The anti-oxidant krill peptide product of bacterium bacillus coagulans, solves the problems, such as that euphausia superba powder is made a low multiple use, simultaneously
Also value-added content of product is substantially increased.Fermented using strain is compounded, trimethylamine is relative in the krill peptide of preparation
Content is 2.49%~5.17%, and without obvious shrimp fishy smell, polypeptide molecular weight is 1000-7000Da, content 60- in product
80%, the TEAC oxidation resistances of product are 60-200 μM of Trolox/mg, and ORAC oxidation resistances are 50-140 μM
Trolox/mg, number of viable reaches 1 × 10 in product6-107CFUs/g。
Compared with prior art, the present invention has advantages below and beneficial effect：
The anti-oxidation peptide of the present invention has higher antioxidation activity, and krill peptide does not have shrimp fishy smell, and has association with probiotics
Same effect；Products taste is good, free from extraneous odour, and is rich in bacillus coagulans, and number of viable reaches 1 × 106-108CFUs/g。
Embodiment
With reference to specific embodiment, the present invention is described in further detail, but embodiments of the present invention are not limited to
This.
The bacterial strain that the present invention uses is bacillus subtilis and bacillus coagulans for bacterial strain；Bacillus subtilis preserving number
CICC10075, Chinese industrial Microbiological Culture Collection administrative center；Bacillus coagulans preserving number CCTCCM2017054, China
Type Tissue Collection.
Embodiment 1
A kind of preparation method of krill anti-oxidation peptide, comprises the following steps：
(1) extracting degreasing euphausia superba powder 100g, deionized water 500g is added, adds degreasing euphausia superba powder and deionized water
The maltose of gross mass 5% mixes, and regulation pH is 6.5, the moist heat sterilization 30min under the conditions of 121 DEG C, is cooled to 35 DEG C, obtains
Euphausia superba powder solution；
(2) peptone 10.0g, beef extract 10.0g, sodium chloride 5.0g are weighed, adds water to be settled to 1000mL, regulation pH to
7.0, obtain fluid nutrient medium；100mL fluid nutrient mediums are taken respectively, 121 DEG C of sterilizing 30min, are cooled to 35 DEG C, are inoculated with withered grass respectively
Bacillus and bacillus coagulans, shaking table culture 20 hours, obtains bacillus subtilis seed under the conditions of 120rpm, 35 DEG C
Liquid and bacillus coagulans seed liquor；The concentration of bacillus subtilis seed liquor miospore is 2.4 × 108Individual/mL and condensation bud
The concentration of spore bacillus seed liquor miospore is 1.6 × 108Individual/ml；
(3) concentration for adjusting bacillus subtilis seed liquor and bacillus coagulans seed liquor miospore is all 108Individual/
ML, then according to volume ratio 1:1.5 are compounded bacillus subtilis seed liquor and bacillus coagulans seed liquor, are answered
With bacterium solution；
(4) inoculum concentration (dosage of compounding bacterium solution be the 5% of euphausia superba powder liquor capacity) of the bacterium solution by 5% will be compounded to connect
Plant in euphausia superba powder solution, then shaker fermentation 35 hours under the conditions of 120rpm, 35 DEG C, under the conditions of 3000r/min
10min is centrifuged, by supernatant in 40 DEG C of spin concentrations to 30Bx, it is anti-oxidant that spray drying obtains the krill rich in probiotics
Peptide.
Krill anti-oxidation peptide manufactured in the present embodiment is reduced to 4.96% without shrimp fishy smell, trimethylamine relative amount, peptide
Content is 68%, and sub- amount is distributed between 2000-5000Da, and TEAC oxidation resistances are 135.1 μM of Trolox/mg, and ORAC resists
Oxidability is 72.4 μM of Trolox/mg, and the quantity of bacillus coagulans reaches 1.5 × 10 in product6CFUs/g。
Embodiment 2
A kind of method for preparing krill anti-oxidation peptide, its step include：
(1) euphausia superba powder 100g is taken, adds deionized water 800g, adds degreasing euphausia superba powder and the total matter of deionized water
The maltose of amount 8% mixes, and regulation pH is 7.0, the moist heat sterilization 30min under the conditions of 121 DEG C, is cooled to 40 DEG C, obtains the South Pole
Krill meal solution；
(2) peptone 10.0g, beef extract 10.0g, sodium chloride 5.0g are weighed, adds water to be settled to 1000mL, regulation pH to
7.0, obtain culture medium；100mL culture mediums are taken respectively, 121 DEG C of sterilizing 30min, are cooled to 40 DEG C, are inoculated with bacillus subtilis respectively
And bacillus coagulans, shaking table culture 24 hours, obtains bacillus subtilis seed liquor and condensation under the conditions of 130rpm, 40 DEG C
Bacillus seed liquor；The concentration of bacillus subtilis seed liquor miospore is 1.9 × 108Individual/mL and bacillus coagulans kind
The concentration of sub- liquid miospore is 1.3 × 108Individual/ml；
(3) concentration for adjusting bacillus subtilis seed liquor and bacillus coagulans seed liquor miospore is all 108Individual/
ML, then according to volume ratio 1:3 are compounded bacillus subtilis seed liquor and bacillus coagulans seed liquor, are compounded
Bacterium solution；
(4) by the strain after compounding, by 8% inoculum concentration, (dosage of compounding bacterium solution is euphausia superba powder liquor capacity
8%) it is inoculated in krill amidin, then shaker fermentation 40 hours under the conditions of 150rpm, 40 DEG C, in 3000r/min bars
10min is centrifuged under part, by supernatant in 40 DEG C of spin concentrations to 40Bx, spray drying obtains the krill rich in probiotics and resisted
Aoxidize peptide.
Krill anti-oxidation peptide finished product manufactured in the present embodiment is reduced to without shrimp fishy smell, trimethylamine relative amount
4.13%, peptide content 71%, for molecular weight distribution between 1000-5000Da, TEAC oxidation resistances are 196.5 μM
Trolox/mg, ORAC oxidation resistance are 123.3 μM of Trolox/mg, in product the quantity of bacillus coagulans reach 1.3 ×
107CFUs/g。
Embodiment 3
A kind of method for preparing krill anti-oxidation peptide, its step include：
(1) euphausia superba powder 100g is taken, adds deionized water 1000g, degreasing euphausia superba powder is added and deionized water is total
The maltose of quality 10% mixes, and regulation pH is 7.0, the moist heat sterilization 30min under the conditions of 121 DEG C, is cooled to 45 DEG C, obtains south
Pole krill meal solution；
(2) peptone 10.0g, beef extract 10.0g, sodium chloride 5.0g are weighed, adds water to be settled to 1000mL, regulation pH to
7.0, obtain culture medium；100mL culture mediums are taken respectively, 121 DEG C of sterilizing 30min, are cooled to 45 DEG C, are inoculated with bacillus subtilis respectively
And bacillus coagulans, shaking table culture 24 hours, obtains bacillus subtilis seed liquor and condensation under the conditions of 150rpm, 45 DEG C
Bacillus seed liquor；The concentration of bacillus subtilis seed liquor miospore is 3.6 × 108Individual/mL and bacillus coagulans kind
The concentration of sub- liquid miospore is 2.3 × 108Individual/ml；
(3) concentration for adjusting bacillus subtilis seed liquor and bacillus coagulans seed liquor miospore is all 108Individual/
ML, then according to volume ratio 1:4 are compounded bacillus subtilis seed liquor and bacillus coagulans seed liquor, are compounded
Bacterium solution；
(4) by the strain after compounding, by 10% inoculum concentration, (dosage of compounding bacterium solution is euphausia superba powder liquor capacity
10%) it is inoculated in krill amidin, then shaker fermentation 45 hours under the conditions of 150rpm, 45 DEG C, in 3000r/min bars
10min is centrifuged under part, by supernatant in 40 DEG C of spin concentrations to 45Bx, spray drying obtains the krill rich in probiotics and resisted
Aoxidize peptide.
Krill anti-oxidation peptide finished product manufactured in the present embodiment is reduced to without shrimp fishy smell, trimethylamine relative amount
3.25%, peptide content 61%, for molecular weight distribution between 3000-7000Da, TEAC oxidation resistances are 73.7 μM
Trolox/mg, ORAC oxidation resistance are 67.6 μM of Trolox/mg, in product the quantity of bacillus coagulans reach 1.1 ×
106CFUs/g。
Claims (10)
- A kind of 1. preparation method of krill anti-oxidation peptide, it is characterised in that：Comprise the following steps：(1) degreasing euphausia superba powder, water and carbon source are mixed, regulation pH value of solution is 6.5~7.5, sterilizing, obtains euphausia superba powder Solution；(2) bacillus subtilis and bacillus coagulans are inoculated in fluid nutrient medium respectively, shaking table is trained under the conditions of 30~45 DEG C 20~24h is supported, the concentration of spore is adjusted, obtains bacillus subtilis seed liquor and bacillus coagulans seed liquor；(3) by bacillus subtilis seed liquor and bacillus coagulans seed liquor according to volume ratio 1:1.5~1:4 are compounded, Obtain compounding bacterium solution；Compounding bacterium solution is mixed with euphausia superba powder solution again, the shaker fermentation 35 under the conditions of 35~45 DEG C ~45 hours, centrifugation, by supernatant concentration, dry, obtain krill anti-oxidation peptide.
- 2. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：It is withered described in step (2) The concentration of careless bacillus seed liquor miospore is 106~109Individual/mL；The concentration of bacillus coagulans seed liquor miospore is 106~109Individual/mL.
- 3. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：It is multiple described in step (3) Spore number ratio with bacillus subtilis in bacterium solution and bacillus coagulans is 1:1.5~1:4.
- 4. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：The mass ratio of degreasing euphausia superba powder and water described in step (1) is 1:5~1:10；The dosage of the carbon source is degreasing south The 5%~10% of the gross mass of pole krill meal and water；The dosage that bacterium solution is compounded described in step (3) is the 5%~10% of euphausia superba powder liquor capacity.
- 5. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：Carbon described in step (1) Source is more than one in maltose, glucose, lactose, sucrose, soluble starch.
- 6. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：Liquid described in step (2) Body culture medium obtains by the following method：Peptone 10.0g is weighed, beef extract 10.0g, sodium chloride 5.0g, adds water to be settled to 1000mL, pH to 6.5~7.5 is adjusted, sterilizing, produces fluid nutrient medium.
- 7. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：Step is gone out described in (1) 25~45min of moist heat sterilization under the conditions of bacterium refers to 110~130 DEG C, cools to 35~45 DEG C after sterilizing；The rotating speed of shaking table culture described in step (2) is 100~200rpm；The rotating speed of shaking table culture described in step (3) is 100~150rpm；Centrifugal rotational speed described in step (3) is 2500~4000r/min；The concentration for 30~45 DEG C of spin concentrations to 30~ 45Bx, the drying are spray drying.
- 8. the preparation method of krill anti-oxidation peptide according to claim 1, it is characterised in that：The bacillus subtilis Preserving number CICC10075, depositary institution：Chinese industrial Microbiological Culture Collection administrative center；Bacillus coagulans preserving number CCTCCM2017054, depositary institution：China typical culture collection center.
- A kind of 9. krill anti-oxidation peptide obtained according to any one of claim 1~8 preparation method.
- 10. the application of krill anti-oxidation peptide according to claim 9, it is characterised in that：The krill is anti-oxidant Peptide is used for health products and/or field of functional food.
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| CN201711275246.XA CN107841518A (en) | 2017-12-06 | 2017-12-06 | A kind of krill anti-oxidation peptide and preparation method and application |
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Cited By (1)
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| CN109182426A (en) * | 2018-09-12 | 2019-01-11 | 荣成泰祥食品股份有限公司 | A method of saline taste peptide is prepared by raw material of krill processing byproduct |
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| CN109182426A (en) * | 2018-09-12 | 2019-01-11 | 荣成泰祥食品股份有限公司 | A method of saline taste peptide is prepared by raw material of krill processing byproduct |
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Application publication date: 20180327 |