CN105255972A - Method for preparing antioxidant active peptide by means of fermenting tilapia skin through aspergillus oryzae - Google Patents
Method for preparing antioxidant active peptide by means of fermenting tilapia skin through aspergillus oryzae Download PDFInfo
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- CN105255972A CN105255972A CN201510776192.XA CN201510776192A CN105255972A CN 105255972 A CN105255972 A CN 105255972A CN 201510776192 A CN201510776192 A CN 201510776192A CN 105255972 A CN105255972 A CN 105255972A
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Abstract
The invention relates to a method for preparing antioxidant active peptide by means of fermenting tilapia skin through aspergillus oryzae and belongs to the technical field of food processing. The tilapia skin serves as a raw material, aspergillus oryzae serves as a fermentation strain to ferment the tilapia skin after pretreatment and extraction, and the tilapia skin antioxidant active peptide is obtained after centrifugation, ultra-filtration and freeze-drying. The fermentation conditions are that aspergillus oryzae serves as the fermentation strain, the addition amount of a seed solution ranges from 1% to 8%, and fermentation is conducted for 50-80 h under the conditions that the temperature ranges from 25 DEG C to 31 DEG C and the pH ranges from 6 to 8. The prepared tilapia skin antioxidant active peptide is remarkable in antioxidant effect, the IC50 values of DPPH free radicals and hydroxyl radicals are 1.09 mg/mL and 1.12 mg/mL respectively, and the tilapia skin is a highly potential bioactive peptide raw material.
Description
Technical field
The present invention relates to a kind of method utilizing aspergillus oryzae fermentation tilapia fishskin to prepare antioxidation active peptides, be specifically related to the preparation method of the significant tilapia fishskin antioxidation active peptides of a kind of antioxidant effect, belong to food processing technology field.
Background technology
Now there are some researches show, the protein content of fish-skin is higher than other positions of fish body, and in fish-skin, collagen content is the highest accounts for more than 80% of its total protein.Especially Java tilapia skin, its protein content reaches 23%, is only second to grass carp skin (29.80%), accounts for 88.5% of its dry weight.So Java tilapia skin is a kind of well collagen protein raw material.And from the research conditions of forefathers, in collagen protein enzymolysis product, at least include antibacterial peptide, ace inhibitory peptide, anti-oxidation peptide etc.These characteristics make collagen protein and hydrolysate thereof have a good application prospect in medical treatment, health care and beauty treatment, also lay a good foundation for preparing anti-oxidation peptide from Java tilapia skin collagen of fish skin enzymolysis solution simultaneously.For this phenomenon, Chinese patent CN101225104 develops the preparation method of tilapia fishskin small molecular peptide chelate zinc salt, but do not relate to the preparation method of antioxidation active peptides, more do not relate to the method that fermentation method prepares tilapia fishskin antioxidation active peptides.Therefore, current fermentation method prepares the research of tilapia fishskin antioxidation active peptides is a new field.
Microbe fermentation method is one of moiety using substrate protein as fermention medium, and select excellent fermented bacterium, the protease hydrolysis substrate protein utilizing microorganism to produce in process of growth prepares biologically active peptides.Microbe fermentation method is prepared biologically active peptides and is had the features such as organic solvent usage quantity is few, efficient.
This patent selects tilapia fishskin to be raw material, and low for processing and utilization rate in current tilapia processing industry, product form is single, and intensive processing and the present situation such as research is few, use biofermentation technique preparation to have the polypeptide of anti-oxidant activity.The collagen protein in fish-skin can not only be recycled, expand the resource of collagen protein and anti-oxidation peptide, widen the range of application of collagen protein, the cleaning (environment-friendly type) realizing the processing fents such as Java tilapia skin utilizes, enrich product structure, improve the capability of sustainable development of China's culture fishery, strengthen the competitive power in world market, shorten and be of great importance at Living marine resources comprehensive utilization technique and theoretic gap with developed country; The relevant blank that fermentation method prepares Java tilapia skin anti-oxidation peptide can also be filled up simultaneously, for the research and development of anti-oxidant series products with accumulate effective reference data as functional foodstuff, medicine and cosmetic and functional factor, there is great economic worth and good social benefit.
Summary of the invention
The present invention is directed to the deficiency that prior art exists, a kind of method utilizing aspergillus oryzae fermentation tilapia fishskin to prepare antioxidation active peptides is provided, fully can meet the demand to antioxidation active peptides on market, to greatest extent for the mankind provide healthy food raw material, and realize the higher value application of tilapia processing fent-fish-skin on this basis.
In order to solve the problem, the technical solution adopted in the present invention is:
Utilize aspergillus oryzae fermentation tilapia fishskin to prepare a method for antioxidation active peptides, comprise the steps:
(1) pre-treatment: the tilapia fishskin after cleaning, after degreasing, soaks 1-2 hour with the hydrochloric acid soln of 0.1-0.13mol/L and the sodium hydroxide solution of 0.1-0.12mol/L successively;
(2) water extraction: pretreated fish-skin is through washing to neutrality, and under 45-50 DEG C of condition, with the distilled water constant temperature homogenate of 1-2 times of volume after 8-10 hour, suction filtration extracts supernatant liquor;
(3) sterilizing: the sterilizing 20 minutes under 121 DEG C of conditions of the supernatant liquor of extraction, then cools for subsequent use;
(4) aspergillus oryzae fermentation: seed liquor addition is 1-8% is 28-31 DEG C, pH in temperature is 6-8 condition bottom fermentation 50-80 hour;
(5) centrifugal ultrafiltration: above-mentioned fermented liquid is got supernatant liquor after the centrifugal 20min of 5000r/min, processes in 0.18MPa, the ultra-filtration membrane that adopts at 40 DEG C molecular weight to be 10kDa, obtains ultrafiltrated;
(6) lyophilize: by ultrafiltrated in-40 ~-50 DEG C, carries out lyophilize under vacuum tightness 0 ~ 20Pa, namely obtains anti-oxidant activity peptide product.
Remarkable advantage of the present invention is:
(1) tilapia fishskin antioxidation active peptides is prepared by biological fermentation process;
(2) the tilapia fishskin antioxidation active peptides antioxidant effect prepared of the present invention is remarkable, and the IC50 value of its DPPH free radical and hydroxy radical qiao is respectively 1.09mg/mL and 1.21mg/mL, is a kind of biologically active peptides raw material of great potential;
(3) biologically active peptides is due to features such as it are safe, easily absorb, cost is low, has good market outlook.The present invention adopts enzymatic hydrolysis rice residue protein to prepare antioxidation active peptides, not only low raw-material cost, and the bioactive peptide obtained have amino acid composition rationally, hypoallergenic, the feature such as safe and reliable, particularly having stronger oxidation-resistance, is a kind of functional food ingredient of great potential;
(4) the present invention is for raw material with tilapia processing fent-fish-skin, a kind of method utilizing aspergillus oryzae fermentation tilapia fishskin to prepare antioxidation active peptides is provided, substantially increase the added value of leftovers of tilapia, and production cost is low, simple process, environmental protection, achieves the higher value application of leftovers of tilapia, has good using value.
Embodiment
Be described in further details the present invention below by embodiment, these embodiments are only used for the present invention is described, do not limit the scope of the invention.
embodiment 1
Utilize aspergillus oryzae fermentation tilapia fishskin to prepare a method for antioxidation active peptides, comprise the steps:
(1) pre-treatment: the tilapia fishskin after cleaning, after degreasing, soaks 1 hour with the hydrochloric acid soln of 0.13mol/L and the sodium hydroxide solution of 0.1mol/L successively;
(2) water extraction: pretreated fish-skin through washing to neutrality, under 50 DEG C of conditions, with the distilled water constant temperature homogenate of 1 times of volume after 8 hours, suction filtration extraction supernatant liquor;
(3) sterilizing: the sterilizing 20 minutes under 121 DEG C of conditions of the supernatant liquor of extraction, then cools for subsequent use;
(4) aspergillus oryzae fermentation: seed liquor addition is 1%, is 28 DEG C in temperature, and pH is 6 condition bottom fermentation 50 hours;
(5) centrifugal ultrafiltration: above-mentioned fermented liquid is got supernatant liquor after the centrifugal 20min of 5000r/min, processes in 0.18MPa, the ultra-filtration membrane that adopts at 40 DEG C molecular weight to be 10kDa, obtains ultrafiltrated;
(6) lyophilize: by ultrafiltrated in-40 DEG C, carries out lyophilize under vacuum tightness 20Pa, namely obtains anti-oxidant activity peptide product.
embodiment 2
Utilize aspergillus oryzae fermentation tilapia fishskin to prepare a method for antioxidation active peptides, comprise the steps:
(1) pre-treatment: the tilapia fishskin after cleaning, after degreasing, soaks 1 hour with the hydrochloric acid soln of 0.13mol/L and the sodium hydroxide solution of 0.1mol/L successively;
(2) water extraction: pretreated fish-skin through washing to neutrality, under 50 DEG C of conditions, with the distilled water constant temperature homogenate of 2 times of volumes after 10 hours, suction filtration extraction supernatant liquor;
(3) sterilizing: the sterilizing 20 minutes under 121 DEG C of conditions of the supernatant liquor of extraction, then cools for subsequent use;
(4) aspergillus oryzae fermentation: seed liquor addition is 8%, is 31 DEG C in temperature, and pH is 8 condition bottom fermentation 80 hours;
(5) centrifugal ultrafiltration: above-mentioned fermented liquid is got supernatant liquor after the centrifugal 20min of 5000r/min, processes in 0.18MPa, the ultra-filtration membrane that adopts at 40 DEG C molecular weight to be 10kDa, obtains ultrafiltrated;
(6) lyophilize: by ultrafiltrated in-50 DEG C, carries out lyophilize under vacuum tightness 5Pa, namely obtains anti-oxidant activity peptide product.
embodiment 3
Take the tilapia fishskin that 900.8g rinses well, 2 hours are soaked with the hydrochloric acid soln of the 0.11mol/L of 700ml and the sodium hydroxide solution of 0.11mol/L successively after skimming treatment, after distilled water wash to neutrality, under 48 DEG C of conditions, with the distilled water constant temperature homogenate 10 hours of 1500ml, suction filtration extracts and obtains supernatant liquor;
Sterilizing: the sterilizing 20 minutes under 121 DEG C of conditions of the supernatant liquor of extraction, then cools for subsequent use;
Aspergillus oryzae ferments: seed liquor addition is 5%, is 28 DEG C in temperature, and pH is 7 condition bottom fermentation 70 hours;
Centrifugal ultrafiltration: above-mentioned fermented liquid is got supernatant liquor after the centrifugal 20min of 5000r/min, processes in 0.18MPa, the ultra-filtration membrane that adopts at 40 DEG C molecular weight to be 10kDa, obtains ultrafiltrated;
Lyophilize: by ultrafiltrated in-45 DEG C, carries out lyophilize under vacuum tightness 10Pa, namely obtains anti-oxidant activity peptide product.
The mensuration of DPPH free radical scavenging activity: the ability removing stable DPPH free radical has been widely used for the resistance of oxidation of testing various polyphenoils.The DPPH radical scavenging activity of the tilapia fishskin tunning of different concns increases along with the increase of concentration.As calculated, when the concentration of tunning reaches 7.7mg/mL, its DPPH free radical scavenging activity can reach 99.91%, shows the radical scavenging activity similar to Vc.
Assess Scavenging activity by IC50 value (antioxidant concentration when clearance rate is 50%), IC50 value lower expression radical scavenging activity is higher.The IC50 value of tilapia fishskin tunning is 1.09mg/mL.
Scavenging activity on hydroxyl free radical measures: the many biochemical reactions occurred in cell are all relevant with hydroxy radical qiao, and it can cause the multiple pathologic such as DNA break, sudden change, lipid oxidation, protein denaturation to change, and hydroxy radical qiao is known most strong oxidizer.The Hydroxyl radical-scavenging ability of tilapia fishskin tunning increases along with the increase of Fermentation Substance Concentration.When Fermentation Substance Concentration is 8.8mg/mL, the Scavenging action to hydroxyl free radical of rice residue protein tunning is 99.78% as calculated, shows the radical scavenging activity similar to Vc.The IC50 value of tilapia fishskin product is 1.21mg/mL.
Claims (1)
1. utilize aspergillus oryzae fermentation tilapia fishskin to prepare a method for antioxidation active peptides, it is characterized in that: comprise the steps:
(1) pre-treatment: the tilapia fishskin after cleaning, after degreasing, soaks 1-2 hour with the hydrochloric acid soln of 0.1-0.13mol/L and the sodium hydroxide solution of 0.1-0.12mol/L successively;
(2) water extraction: pretreated fish-skin is through washing to neutrality, and under 45-50 DEG C of condition, with the distilled water constant temperature homogenate of 1-2 times of volume after 8-10 hour, suction filtration extracts supernatant liquor;
(3) sterilizing: the sterilizing 20 minutes under 121 DEG C of conditions of the supernatant liquor of extraction, then cools for subsequent use;
(4) aspergillus oryzae fermentation: seed liquor addition is 1-8% is 28-31 DEG C, pH in temperature is 6-8 condition bottom fermentation 50-80 hour;
(5) centrifugal ultrafiltration: above-mentioned fermented liquid is got supernatant liquor after the centrifugal 20min of 5000r/min, processes in 0.18MPa, the ultra-filtration membrane that adopts at 40 DEG C molecular weight to be 10kDa, obtains ultrafiltrated;
(6) lyophilize: by ultrafiltrated in-40 ~-50 DEG C, carries out lyophilize under vacuum tightness 0 ~ 20Pa, namely obtains anti-oxidant activity peptide product.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105861565A (en) * | 2016-06-04 | 2016-08-17 | 中国海洋大学 | Turbot fishskin fermentation product with antioxidant activity |
| CN110074356A (en) * | 2019-06-04 | 2019-08-02 | 江南大学 | A kind of Java tilapia skin deliming degreasing method |
| WO2021205851A1 (en) * | 2020-04-07 | 2021-10-14 | 新田ゼラチン株式会社 | Peptide composition and method for producing same |
| CN114539388A (en) * | 2022-01-20 | 2022-05-27 | 大连民族大学 | Method for preparing salmon skin collagen by adopting microbial fermentation method |
| CN115413241A (en) * | 2020-04-07 | 2022-11-29 | 新田明胶株式会社 | Biological function regulator, epidermal metabolism promoter, lipopexia inhibitor, lipolysis promoter, adiponectin production promoter, functional food, cosmetic, and method for producing biological function regulator |
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| CN102199646A (en) * | 2011-03-15 | 2011-09-28 | 广东海洋大学 | Method for preparing collagen peptide with fish skin |
| CN105255982A (en) * | 2015-11-13 | 2016-01-20 | 周金全 | Method for preparing antioxidant active peptide by means of fermenting tilapia skin through aspergillus oryzae |
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| CN102199646A (en) * | 2011-03-15 | 2011-09-28 | 广东海洋大学 | Method for preparing collagen peptide with fish skin |
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Cited By (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105861565A (en) * | 2016-06-04 | 2016-08-17 | 中国海洋大学 | Turbot fishskin fermentation product with antioxidant activity |
| CN105861565B (en) * | 2016-06-04 | 2019-08-23 | 中国海洋大学 | A kind of turbot fish-skin tunning with antioxidant activity |
| CN110074356A (en) * | 2019-06-04 | 2019-08-02 | 江南大学 | A kind of Java tilapia skin deliming degreasing method |
| WO2021205851A1 (en) * | 2020-04-07 | 2021-10-14 | 新田ゼラチン株式会社 | Peptide composition and method for producing same |
| CN115413241A (en) * | 2020-04-07 | 2022-11-29 | 新田明胶株式会社 | Biological function regulator, epidermal metabolism promoter, lipopexia inhibitor, lipolysis promoter, adiponectin production promoter, functional food, cosmetic, and method for producing biological function regulator |
| CN115426895A (en) * | 2020-04-07 | 2022-12-02 | 新田明胶株式会社 | Peptide composition and method for producing same |
| CN114539388A (en) * | 2022-01-20 | 2022-05-27 | 大连民族大学 | Method for preparing salmon skin collagen by adopting microbial fermentation method |
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Application publication date: 20160120 |