CN107841519A - 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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- CN107841519A CN107841519A CN201711275248.9A CN201711275248A CN107841519A CN 107841519 A CN107841519 A CN 107841519A CN 201711275248 A CN201711275248 A CN 201711275248A CN 107841519 A CN107841519 A CN 107841519A
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- China
- Prior art keywords
- krill
- oxidation peptide
- bacillus subtilis
- preparation
- euphausia superba
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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 7.0~7.5, sterilizing, obtains euphausia superba powder solution；(2) bacillus subtilis is inoculated in fluid nutrient medium, 20~24h of shaking table culture under the conditions of 29~31 DEG C, obtains bacillus subtilis seed liquor；(3) bacillus subtilis seed liquor is mixed with euphausia superba powder solution, shaker fermentation 35~42 hours under the conditions of 30~40 DEG C, centrifuged, by supernatant concentration, dried, obtain krill anti-oxidation peptide.The method of the present invention is simple, environmental protection；And the molecular weight of peptide has higher antioxidation activity between 1000 7000Da；Rich in vitamin B12, for cosmetics, health products and/or field of functional food.
Description
Technical field
The invention belongs to biological fermentation engineering field, is related to a kind of with fermentation of bacillus subtilis processing krill acquisition
A kind of method of anti-oxidation peptide rich in vitamin B12, and in particular to krill anti-oxidation 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.Therefore, it is to improve krill resource to find suitable process and krill albumen is processed
The key of comprehensive utilization.
Krill protein degradation is thing active peptide using caused protease in fermentation process by the present invention, and this measure is not
The added value of krill albumen can be only greatly improved, a large amount of high-quality raw materials is provided for health products trade, can also be krill egg
A new approach is opened up in white deep processing.
The content of the invention
It is an object of the invention to provide a kind of method that krill anti-oxidation peptide is prepared using fermentation of bacillus subtilis,
So as to improve the comprehensive utilization ratio of krill protein and 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 cosmetic field, health products and/or field of functional food, for preparing cosmetics, health products and/or feature
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 7.0~7.5, sterilizing, obtains South Pole phosphorus
Shrimp med solution；
(2) bacillus subtilis is inoculated in fluid nutrient medium, 20~24h of shaking table culture under the conditions of 29~31 DEG C, obtained
Bacillus subtilis seed liquor；
(3) bacillus subtilis seed liquor is mixed with euphausia superba powder solution, the shaking table under the conditions of 30~40 DEG C
Fermentation 35~42 hours, centrifugation, by supernatant concentration, dry, obtain krill anti-oxidation peptide.
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 4%~8% of fat euphausia superba powder and the gross mass of water；Degreasing krill meal is remaining leftover bits and pieces after the extracted krill oil of krill meal
Material.
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), 30 are cooled to after sterilizing
~40 DEG C；
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 7.0~7.5, 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 30~40 DEG C after sterilizing；
The concentration of bacillus subtilis seed liquor miospore described in step (2) is 106~109Individual/mL；
The rotating speed of shaking table culture described in step (2) is 100~200rpm；
The dosage of bacillus subtilis seed liquor 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.
Degreasing euphausia superba powder is rich and easy to get used by invention, and the above method can prepare high anti-oxidation krill
Peptide, solve the problems, such as that euphausia superba powder is made a low multiple use, while also substantially increase value-added content of product.Utilize withered grass bud
Spore bacillus is fermented, and the molecular weight of peptide is between 1000-7000Da in the krill peptide product of preparation, content 50-
80%, TEAC oxidation resistance are 80-180 μM of Trolox/mg, and ORAC oxidation resistances are 50-120 μM of Trolox/mg, dimension
Raw plain B12 content is 9.3-17.5 μ g/kg.Effective adding ingredient can be used as to be used among health products.
The present invention is prepared with anti-by caused proteasome degradation degreasing krill meal during fermentation of bacillus subtilis
The krill peptide of oxidation activity, finally give the product of krill peptide
Compared with prior art, the present invention has advantages below and beneficial effect：
The krill anti-oxidation peptide preparation method of the present invention is simple, environmental protection；And the molecular weight of peptide 1000-7000Da it
Between, content 50-80%, there is higher antioxidation activity；Vitamin B12 is rich in product, content is 9.3-17.5 μ g/
kg.The present invention not only increases the utilization of the synthesis of krill resource, and is prepared by the fermentation of microorganism rich in vitamin
B12 krill peptide, can be as the main adding ingredient of health products.
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 10075 for bacterial strain, bacillus subtilis preserving number
CICC10075, Chinese industrial Microbiological Culture Collection administrative center.
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 4% mixes, and regulation pH is 7.0, the moist heat sterilization 30min under the conditions of 121 DEG C, is cooled to 30 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, 121 DEG C of sterilizing 30min, 30 DEG C is cooled to, is inoculated with bacillus subtilis
Bacterium, shaking table culture 20 hours, obtains bacillus subtilis seed liquor under the conditions of 120rpm, 30 DEG C；Bacillus subtilis seed
The concentration of liquid miospore is 1.9 × 108Individual/ml；
(3) by bacillus subtilis seed liquor, by 5% inoculum concentration, (dosage of bacillus subtilis seed liquor is South Pole phosphorus
5%) shrimp med liquor capacity is inoculated in euphausia superba powder solution, then shaker fermentation 35 is small under the conditions of 120rpm, 30 DEG C
When, 10min is centrifuged under the conditions of 3000r/min, supernatant is obtained into South Pole phosphorus in 40 DEG C of spin concentrations to 35Bx, spray drying
Shrimp anti-oxidation peptide.
Krill peptide powder manufactured in the present embodiment, the content of peptide are 57%, molecular weight distribution 5000-7000Da it
Between, TEAC oxidation resistances are 87.6 μM of Trolox/mg, and ORAC oxidation resistances are 52.3 μM of Trolox/mg, vitamin
B12 contents are 9.36 μ g/kg.
Embodiment 2
A kind of preparation method of krill anti-oxidation peptide, comprises the following steps：
(1) extracting degreasing euphausia superba powder 100g, deionized water 800g is added, adds degreasing euphausia superba powder and deionized water
The maltose of gross mass 6% mixes, and regulation pH is 7.5, the moist heat sterilization 30min under the conditions of 121 DEG C, is cooled to 30 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
7.2, obtain culture medium；100mL culture mediums are taken, 121 DEG C of sterilizing 30min, 30 DEG C is cooled to, is inoculated with bacillus subtilis,
130rpm, shaking table culture 24 hours under the conditions of 30 DEG C, obtain bacillus subtilis seed liquor；Spore in bacillus subtilis seed liquor
The concentration of son is 2.3 × 108Individual/ml；
(3) by bacillus subtilis seed liquor, by 8% inoculum concentration, (dosage of bacillus subtilis seed liquor is South Pole phosphorus
8%) shrimp med liquor capacity is inoculated in euphausia superba powder solution, then shaker fermentation 40 is small under the conditions of 150rpm, 30 DEG C
When, 10min is centrifuged under the conditions of 3000r/min, supernatant is obtained into South Pole phosphorus in 40 DEG C of spin concentrations to 40Bx, spray drying
Shrimp anti-oxidation peptide.
Krill peptide powder manufactured in the present embodiment, the content of peptide are 72%, molecular weight distribution in 3000-6000Da,
TEAC oxidation resistances are 140.5 μM of Trolox/mg, and ORAC oxidation resistances are 100.2 μM of Trolox/mg, vitamin B12
Content is 16.58 μ g/kg.
Embodiment 3
A kind of preparation method of krill anti-oxidation peptide, comprises the following steps：
(1) extracting degreasing euphausia superba powder 100g, deionized water 1000g is added, adds degreasing euphausia superba powder and deionization
The maltose of water gross mass 8% mixes, and regulation pH is 7.5, the moist heat sterilization 30min under the conditions of 121 DEG C, is cooled to 30 DEG C, obtains
To 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
7.5, obtain culture medium；100mL culture mediums are taken, 121 DEG C of sterilizing 30min, 30 DEG C is cooled to, is inoculated with bacillus subtilis,
150rpm, shaking table culture 24 hours under the conditions of 30 DEG C, obtain bacillus subtilis seed liquor；Spore in bacillus subtilis seed liquor
The concentration of son is 2.9 × 108Individual/ml；
(3) by bacillus subtilis seed liquor, by 10% inoculum concentration, (dosage of bacillus subtilis seed liquor is the South Pole
10%) krill meal liquor capacity is inoculated in euphausia superba powder solution, then the shaker fermentation 42 under the conditions of 150rpm, 30 DEG C
Hour, 10min is centrifuged under the conditions of 3000r/min, supernatant is obtained into the South Pole in 40 DEG C of spin concentrations to 45Bx, spray drying
Krill anti-oxidation peptide.
Krill peptide powder manufactured in the present embodiment, the content of peptide is 80%, and molecular weight distribution is in 1000-4000Da
TEAC oxidation resistances are 180.8 μM of Trolox/mg, and ORAC oxidation resistances are 120.3 μM of Trolox/mg, vitamin B12
Content is 14.79 μ g/kg.
Claims (9)
- 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 7.0~7.5, sterilizing, obtains euphausia superba powder Solution；(2) bacillus subtilis is inoculated in fluid nutrient medium, 20~24h of shaking table culture under the conditions of 29~31 DEG C, obtains withered grass Bacillus seed liquor；(3) bacillus subtilis seed liquor is mixed with euphausia superba powder solution, the shaker fermentation under the conditions of 30~40 DEG C 35~42 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：The concentration of bacillus subtilis seed liquor miospore described in step (2) is 106~109Individual/mL.
- 3. 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 4%~8% of the gross mass of pole krill meal and water；The dosage of bacillus subtilis seed liquor described in step (3) is the 5%~10% of euphausia superba powder liquor capacity.
- 4. 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.
- 5. 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 7.0~7.5 is adjusted, sterilizing, produces fluid nutrient medium.
- 6. 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 30~40 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.
- 7. 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.
- A kind of 8. krill anti-oxidation peptide obtained according to any one of claim 1~7 preparation method.
- 9. the application of krill anti-oxidation peptide according to claim 8, it is characterised in that：The krill anti-oxidation peptide For cosmetics, health products and/or field of functional food.
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| CN201711275248.9A CN107841519A (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)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112195212A (en) * | 2020-10-15 | 2021-01-08 | 华中农业大学 | Method for preparing bee pupa antioxidant polypeptide by microbial fermentation |
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| JP2013523100A (en) * | 2010-12-14 | 2013-06-17 | 大▲連▼工▲業▼大学 | Method for producing krill oil or krill concentrate, krill meal from nankoku krill |
| CN106244484A (en) * | 2016-08-09 | 2016-12-21 | 中国水产科学研究院黄海水产研究所 | Antarctic krill fungal component, Antarctic krill anti-oxidation peptide and its preparation method and application |
-
2017
- 2017-12-06 CN CN201711275248.9A patent/CN107841519A/en active Pending
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP2013523100A (en) * | 2010-12-14 | 2013-06-17 | 大▲連▼工▲業▼大学 | Method for producing krill oil or krill concentrate, krill meal from nankoku krill |
| CN106244484A (en) * | 2016-08-09 | 2016-12-21 | 中国水产科学研究院黄海水产研究所 | Antarctic krill fungal component, Antarctic krill anti-oxidation peptide and its preparation method and application |
Non-Patent Citations (6)
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| JIANAN SUN,ET AL.: "An environmental friendly process for Antarctic krill (Euphausia superba) utilization using fermentation technology", 《JOURNAL OF CLEANER PRODUCTION》 * |
| 刘文珍: "水产养殖中微生物的应用及研究进展", 《水产科学》 * |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112195212A (en) * | 2020-10-15 | 2021-01-08 | 华中农业大学 | Method for preparing bee pupa antioxidant polypeptide by microbial fermentation |
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