CN104707128A - Method for preparing anti-oxidation active liquid by performing enzymatic hydrolysis on soft-shelled turtle eggs - Google Patents
Method for preparing anti-oxidation active liquid by performing enzymatic hydrolysis on soft-shelled turtle eggs Download PDFInfo
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- CN104707128A CN104707128A CN201510150779.XA CN201510150779A CN104707128A CN 104707128 A CN104707128 A CN 104707128A CN 201510150779 A CN201510150779 A CN 201510150779A CN 104707128 A CN104707128 A CN 104707128A
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Abstract
The invention discloses a method for preparing anti-oxidation active liquid by performing enzymatic hydrolysis on soft-shelled turtle eggs. The method comprises the following steps: reverse osmosis water washing, stirring, passivation, filtration, centrifugation, concentration, membrane filtration, filling and sealing. According to the technical scheme of the method, soft-shelled turtle egg protein is mildly hydrolyzed by using a modern biological enzyme engineering technology, hydrolysis components are relatively stable, environmental pollution cannot be caused, and the method is easy to operate and control; by controlling hydrolysis conditions, nutritional values of the anti-oxidation active liquid can be improved and the anti-oxidation active liquid can be easily absorbed; moreover, the mixed liquid has a certain antioxidant capacity, and the antioxidant activity of the mixed liquid cannot be reduced after the mixed liquid is heated for 30 minutes ( by virtue of pasteurization) in hot water of which the pH value is 4.0 and the temperature is 85 DEG C; and the mixed liquid of amino acids, peptides and relatively large-molecular protein, prepared by using the method disclosed by the invention, have antioxidant capacity and thermal stability and can be applied to the field of processing of cosmetics, health products and foods.
Description
Technical field
The present invention relates to biological technical field, be specifically related to a kind of method of Trionyx sinensis Wiegmann egg being carried out enzymatic hydrolysis and preparing antioxidant activity liquid.
Background technology
Breeding soft-shell turtle enterprise general working procedure is such: 5-8 month, and Trionyx sinensis Wiegmann starts to lay eggs, and Trionyx sinensis Wiegmann egg is just selected, to distinguish germ cell and unfertilized egg after producing.Germ cell continues hatching, until hatch Trionyx sinensis Wiegmann Seedling, sells Trionyx sinensis Wiegmann Seedling or continues cultivation and to grow up sale finished product.And at present, unfertilized turtle egg is generally processed as raw-food material or abandoned, cause and waste significantly.Effectively can suppress the oxidation reaction of free radical when antioxidant refers to that low concentration exists, and eliminate the murder by poisoning of radical pair human body, be the anti-aging in advance major reasons of people.Trionyx sinensis Wiegmann egg is not also utilized to prepare the relevant report of antioxidant at present.
Summary of the invention
The object of the invention is to provide a kind of method of Trionyx sinensis Wiegmann egg being carried out enzymatic hydrolysis and preparing antioxidant activity liquid.
The object of the invention is to be achieved through the following technical solutions:
Trionyx sinensis Wiegmann egg is carried out the method that enzymatic hydrolysis prepares antioxidant activity liquid, it is characterized in that, comprise the steps:
(1) reverse osmosis water cleaning: use reverse osmosis water to clean Trionyx sinensis Wiegmann egg, reverse osmosis water is obtained by the reverse osmosis water equipment that market is general, requires that electrical conductivity is less than 5 s/cm;
(2) stir: put into bioreactor after being shelled by Trionyx sinensis Wiegmann egg, add the deionized water of more than 15 times of Trionyx sinensis Wiegmann egg size amount, add enzyme and carry out enzymatic reaction, and then stir, rotating speed 500-4500rpm;
(3) passivation: improve the reaction temperature in bioreactor, be warming up to 75-98 DEG C, time 30-50 minute, reach the object that bioenzyme activity stops;
(4) filter: the liquid after passivation is adopted common Bag filter, removing solid content;
(5) centrifugal: to adopt a small amount of oils and fats in disc centrifuge separating liquid, rotating speed 3000-6000rpm;
(6) concentrated: to adopt vacuum concentration, more than vacuum 600mm mercury column, temperature 50-75 DEG C, is concentrated into the 3-10 of Trionyx sinensis Wiegmann egg size amount doubly;
(7) membrane filtration: molecular film filtration is carried out to liquid concentrated after hydrolysis, carries out 3kda, 10kda, 50kda molecular filtration respectively, obtain aminoacid, peptide class and more macromolecular mixed liquid of protein;
(8) filling and sealing: with vial filling and sealing, sterilization, sterilization temperature 65-95 DEG C, time 30-45 minute, is cooled to room temperature with tap water.
The enzyme added in described step (2) is divided into four kinds: 1, neutral protease, by the proteolytic enzyme extracted after fermentation of bacillus subtilis, preference temperature 35-55 DEG C, PH6-7.5, vigor 100,000 unit every gram, addition 0.05-0.25%, action time 2-3 hour, utilize the enzymatic reaction of neutral protease, the macro-molecular protein of Trionyx sinensis Wiegmann egg is hydrolyzed into small-molecular peptides and aminoacid; 2, the yellow and white specific enzyme, preference temperature 50-55 DEG C, PH7-8, action time 2-3 hour, addition 0.1-0.3%; 3, alkaline protease, is the proteolytic enzyme that the lichen bacillus ferments produces, the high molecular weight protein of Trionyx sinensis Wiegmann egg is hydrolyzed to peptide and aminoacid; Preference temperature 25-70 DEG C, PH7-10, action time 2-3 hour, addition 0.1-0.2%; 4, pancreatin, is extracted by animal pancreas, preference temperature 50-55 DEG C, PH7-8, time 2-3 hour, addition 0.05-0.20%; The addition of described enzyme all with the weight of Trionyx sinensis Wiegmann egg for benchmark.
Beneficial effect of the present invention: technical solution of the present invention application modern biotechnology enzyme engineering technology, mild hydrolysis turtle egg albumen, hydrolysing component is relatively stable, environmentally safe, easily manipulates.By the control of hydrolysising condition, nutritive value gets a promotion, and easily absorbs.And this mixing material has certain oxidation resistance, the antioxidant activity of this mixing material heats 30 minutes (pasteurize) activity and does not reduce in the hot water of PH4.0 and 85 DEG C.The aminoacid, peptide class and the more macromolecular mixed liquid of protein that are obtained by this method, owing to having oxidation resistance and heat stability, can be applicable to cosmetics, health product, food processing field.
Detailed description of the invention
The following examples can illustrate in greater detail the present invention, but do not limit the present invention in any form.
Embodiment 1:
(1) reverse osmosis water cleaning: use reverse osmosis water to clean Trionyx sinensis Wiegmann egg, reverse osmosis water is obtained by the reverse osmosis water equipment that market is general, requires that electrical conductivity is less than 5 s/cm;
(2) stir: after being shelled by Trionyx sinensis Wiegmann egg, put into bioreactor, add the deionized water of more than 15 times of Trionyx sinensis Wiegmann egg size amount, add enzyme and carry out enzymatic reaction, add enzyme and be divided into four kinds: 1, neutral protease, by the proteolytic enzyme extracted after fermentation of bacillus subtilis, preference temperature 35-55 DEG C, PH6-7.5, vigor 100,000 unit every gram, addition 0.05-0.25%, action time, 2-3 hour, utilized the enzymatic reaction of neutral protease, and the macro-molecular protein of Trionyx sinensis Wiegmann egg is hydrolyzed into small-molecular peptides and aminoacid; 2, the yellow and white specific enzyme, preference temperature 50-55 DEG C, PH7-8, action time 2-3 hour, addition 0.1-0.3%; 3, alkaline protease, is the proteolytic enzyme that the lichen bacillus ferments produces, the high molecular weight protein of Trionyx sinensis Wiegmann egg is hydrolyzed to peptide and aminoacid; Preference temperature 25-70 DEG C, PH7-10, action time 2-3 hour, addition 0.1-0.2%; 4, pancreatin, is extracted by animal pancreas, preference temperature 50-55 DEG C, PH7-8, time 2-3 hour, addition 0.05-0.20%; The addition percent of described four kinds of enzyme all with the weight of Trionyx sinensis Wiegmann egg for benchmark; And then stir, rotating speed 500-4500rpm;
(3) passivation: improve the reaction temperature in bioreactor, be warming up to 75-98 DEG C, time 30-50 minute, reach the object that bioenzyme activity stops;
(4) filter: the liquid after passivation is adopted common Bag filter, removing solid content;
(5) centrifugal: to adopt a small amount of oils and fats in disc centrifuge separating liquid, rotating speed 3000-6000rpm;
(6) concentrated: to adopt vacuum concentration, more than vacuum 600mm mercury column, temperature 50-75 DEG C, is concentrated into the 3-10 of Trionyx sinensis Wiegmann egg size amount doubly;
(7) membrane filtration: molecular film filtration is carried out to liquid concentrated after hydrolysis, carries out 3kda, 10kda, 50kda molecular filtration respectively, obtain aminoacid, peptide class and more macromolecular mixed liquid of protein;
(8) filling and sealing: with vial filling and sealing, sterilization, sterilization temperature 65-95 DEG C, time 30-45 minute, is cooled to room temperature with tap water.
Claims (2)
1. Trionyx sinensis Wiegmann egg is carried out the method that enzymatic hydrolysis prepares antioxidant activity liquid, it is characterized in that, comprise the steps:
(1) reverse osmosis water cleaning: use reverse osmosis water to clean Trionyx sinensis Wiegmann egg, reverse osmosis water is obtained by the reverse osmosis water equipment that market is general, requires that electrical conductivity is less than 5 s/cm;
(2) stir: put into bioreactor after being shelled by Trionyx sinensis Wiegmann egg, add the deionized water of more than 15 times of Trionyx sinensis Wiegmann egg size amount, add enzyme and carry out enzymatic reaction, and then stir, rotating speed 500-4500rpm;
(3) passivation: improve the reaction temperature in bioreactor, be warming up to 75-98 DEG C, time 30-50 minute, reach the object that bioenzyme activity stops;
(4) filter: the liquid after passivation is adopted common Bag filter, removing solid content;
(5) centrifugal: to adopt a small amount of oils and fats in disc centrifuge separating liquid, rotating speed 3000-6000rpm;
(6) concentrated: to adopt vacuum concentration, more than vacuum 600mm mercury column, temperature 50-75 DEG C, is concentrated into the 3-10 of Trionyx sinensis Wiegmann egg size amount doubly;
(7) membrane filtration: molecular film filtration is carried out to liquid concentrated after hydrolysis, carries out 3kda, 10kda, 50kda molecular filtration respectively, obtain aminoacid, peptide class and more macromolecular mixed liquid of protein;
(8) filling and sealing: with vial filling and sealing, sterilization, sterilization temperature 65-95 DEG C, time 30-45 minute, is cooled to room temperature with tap water.
2. a kind of method of Trionyx sinensis Wiegmann egg being carried out enzymatic hydrolysis and preparing antioxidant activity liquid according to claim 1, it is characterized in that, the enzyme added in described step (2) is divided into four kinds: 1, neutral protease, by the proteolytic enzyme extracted after fermentation of bacillus subtilis, preference temperature 35-55 DEG C, PH6-7.5, vigor 100,000 unit every gram, addition 0.05-0.25%, action time 2-3 hour, utilize the enzymatic reaction of neutral protease, the macro-molecular protein of Trionyx sinensis Wiegmann egg is hydrolyzed into small-molecular peptides and aminoacid; 2, the yellow and white specific enzyme, preference temperature 50-55 DEG C, PH7-8, action time 2-3 hour, addition 0.1-0.3%; 3, alkaline protease, is the proteolytic enzyme that the lichen bacillus ferments produces, the high molecular weight protein of Trionyx sinensis Wiegmann egg is hydrolyzed to peptide and aminoacid; Preference temperature 25-70 DEG C, PH7-10, action time 2-3 hour, addition 0.1-0.2%; 4, pancreatin, is extracted by animal pancreas, preference temperature 50-55 DEG C, PH7-8, time 2-3 hour, addition 0.05-0.20%; The addition of described enzyme all with the weight of Trionyx sinensis Wiegmann egg for benchmark.
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Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105238545A (en) * | 2015-09-23 | 2016-01-13 | 集美大学 | Preparation method for combined production of micromolecular soft-shelled turtle peptide and soft-shelled turtle oil |
| CN105997823A (en) * | 2016-07-14 | 2016-10-12 | 湖南湘湖水产有限公司 | Turtle oil and egg white beauty mask |
| CN107177657A (en) * | 2017-07-02 | 2017-09-19 | 常德至善生物科技有限公司 | A kind of method that step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid |
| CN107201390A (en) * | 2017-07-02 | 2017-09-26 | 常德至善生物科技有限公司 | A kind of method that stepwise discretization method hydrolysis soft-shelled turtle egg prepares high anti-oxidation active peptide liquid |
| CN108251484A (en) * | 2018-03-21 | 2018-07-06 | 杭州早稻田生物技术有限公司 | Soft-shelled turtle active peptide albumen powder and preparation method thereof |
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| CN1994328A (en) * | 2006-01-05 | 2007-07-11 | 李勇 | Use of Chinese softed turtle egg in preparation of medicine or food for relieving diabetic symptoms and lowering blood sugar |
| CN101991035A (en) * | 2010-11-23 | 2011-03-30 | 王鸿泰 | Nutrient fluid prepared from animal embryo |
| TWM448275U (en) * | 2012-08-16 | 2013-03-11 | Qiu-Qi Chen | Soft-shell turtle egg capsule structure |
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| CN1994328A (en) * | 2006-01-05 | 2007-07-11 | 李勇 | Use of Chinese softed turtle egg in preparation of medicine or food for relieving diabetic symptoms and lowering blood sugar |
| CN101991035A (en) * | 2010-11-23 | 2011-03-30 | 王鸿泰 | Nutrient fluid prepared from animal embryo |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105238545A (en) * | 2015-09-23 | 2016-01-13 | 集美大学 | Preparation method for combined production of micromolecular soft-shelled turtle peptide and soft-shelled turtle oil |
| CN105997823A (en) * | 2016-07-14 | 2016-10-12 | 湖南湘湖水产有限公司 | Turtle oil and egg white beauty mask |
| CN107177657A (en) * | 2017-07-02 | 2017-09-19 | 常德至善生物科技有限公司 | A kind of method that step fermentation soft-shelled turtle egg prepares anti-oxidation peptide liquid |
| CN107201390A (en) * | 2017-07-02 | 2017-09-26 | 常德至善生物科技有限公司 | A kind of method that stepwise discretization method hydrolysis soft-shelled turtle egg prepares high anti-oxidation active peptide liquid |
| CN107201390B (en) * | 2017-07-02 | 2021-01-08 | 山东大树达孚特膳食品有限公司 | Method for preparing high-antioxidant active peptide liquid by hydrolyzing soft-shelled turtle eggs through stepwise enzymolysis method |
| CN108251484A (en) * | 2018-03-21 | 2018-07-06 | 杭州早稻田生物技术有限公司 | Soft-shelled turtle active peptide albumen powder and preparation method thereof |
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