CN104694606B - A kind of method that small-molecular peptides are prepared with complex enzyme for hydrolyzing Oyster Protein - Google Patents
A kind of method that small-molecular peptides are prepared with complex enzyme for hydrolyzing Oyster Protein Download PDFInfo
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- CN104694606B CN104694606B CN201510144750.0A CN201510144750A CN104694606B CN 104694606 B CN104694606 B CN 104694606B CN 201510144750 A CN201510144750 A CN 201510144750A CN 104694606 B CN104694606 B CN 104694606B
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Abstract
The present invention relates to the preparing technical field of small-molecular peptides, more particularly to a kind of method that small-molecular peptides are prepared with complex enzyme for hydrolyzing Oyster Protein, is specially：(1) oyster is cleaned, adds water and ficin, digested, take oyster meat；(2) oyster meat is smashed, adds water, stir evenly, obtain oyster slurries；(3) complex enzyme is added, is digested, enzyme deactivation, obtains enzymolysis liquid；(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.The method of the present invention technique is simple, easily operated, digests mechanical dejacketing, saves the time, shorten the production cycle, and the protein peptides being prepared are small-molecular peptides, and molecular weight is small, and activity is high.
Description
Technical field
It is more particularly to a kind of to be prepared with complex enzyme for hydrolyzing Oyster Protein the present invention relates to the preparing technical field of small-molecular peptides
The method of small-molecular peptides.
Background technology
Oyster (Oyster) is a kind of marine shellfish.Oyster belongs to Mollusca in classification, lamellibranchiata, Anisomyaria,
Oyster superfamily, Ostreidae, it is a kind of bivalve marine product of food and medicament dual-purpose.There is cultivation China north and south is coastal, particularly
Guangdong Province developed comparatively fast in recent years, and oyster meat is milky white, delicate, full of nutrition, except containing abundant protein, vitamin and
Outside carbohydrate, also containing more than ten kinds of amino acid, mineral nutrients needed by human etc..
Recent studies shows：Oyster is containing 18 kinds of amino acid, liver glycogen, B family vitamin, taurine and calcium, phosphorus, iron, zinc etc.
Nutritional ingredient, immunity of organisms can be improved by often eating, its contained taurine can reducing blood lipid, blood pressure lowering.Clinically, oyster is being treated
Insomnia, chronic otitis media, children's ephidrosis, fibroid and infantile enuresis etc. have the effect of prominent, and (traditional Chinese medicine is believed
Breath, 2011,28 (1) 114-116.).Oyster extract has antibacterial action, has suppression to poliovirus and influenza virus
Effect, its water soluble ingredient still improve animal body immunity etc..
At present, the method for someone's enzymolysis prepares protein peptides and lives, as Chinese invention patent Publication No. CN1680578A is public
A kind of preparation method of oyster active peptides is opened, using Pacific oyster as raw material, using protease hydrolytic Oyster Protein, then with receiving
Filter technology desalination and concentration, can obtain oyster active peptides.Used in this method for protease, and the selection of enzyme is production active peptide
Key, the hydrolysis ability of enzyme has specificity, if wanting to obtain the polypeptide with unique activity, a kind of alone enzyme effect is bad.
Also the method for someone's complex enzyme zymohydrolysis prepares protein peptides, as Chinese invention patent Publication No. CN102250997B discloses one
The method that kind prepares active peptide with complex enzyme for hydrolyzing Oyster Protein, is that oyster is shelled, smashs to pieces, add distilled water, is homogenized, enzymolysis,
Enzyme deactivation, adjusts pH value, after vacuum decompression is filtered and is freeze-dried, obtains the active peptide that a kind of molecular weight is less than 3000Da.The party
Although method prepares protein peptides, the protein peptides that the combination of selected enzyme prepares, molecular weight using the method for complex enzyme zymohydrolysis
It is opposite or bigger.
The content of the invention
The technical problems to be solved by the invention are to provide a kind of prepares small-molecular peptides with complex enzyme for hydrolyzing Oyster Protein
Method, this method technique is simple, easily operated, and the small-molecular peptides molecular weight being prepared is small.
To achieve the above object, the present invention uses following technical scheme：
A kind of method that small-molecular peptides are prepared with complex enzyme for hydrolyzing Oyster Protein, including following preparation process：
(1) oyster is cleaned, add oyster 7~10 times of water of weight, add oyster weight 0.01~0.05% without flower
Fruit protease, adjusts pH value to 4.7~4.9, is warming up to 51~54 DEG C, digests 5~10min, take oyster meat；
(2) oyster meat is smashed, adds 2~4 times of water of its weight, stir evenly, obtain oyster slurries；
(3) complex enzyme of oyster meat weight 0.04~0.09% is added, adjusts pH to 5.3~6.5, rise temperature to 46
~52 DEG C, 30~50min of homogeneous, enzyme deactivation, obtains enzymolysis liquid；
(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.
Complex enzyme of the present invention is the mixture of ficin, pronase and subtilopeptidase A.
Whole protein can be hydrolyzed to small-molecular peptides by the enzyme effect site used in the present invention without specificity.
Homogenization pressure is 16~21Mpa in step (3) of the present invention.Homogeneous makes oyster slurries micronized, homogenization, reduces and divides
Dissipate thing scale and improve dispersion distributing homogeneity.The present invention provides optimal homogenization pressure condition.
The temperature of enzyme deactivation is improved to 75~80 DEG C in step (3) of the present invention.The enzyme-removal temperature of the present invention is gentle, can either be complete
Full enzyme deactivation, does not lose the small-molecular peptides amount of extraction yet.
The weight ratio of ficin of the present invention, pronase and subtilopeptidase A is 4:2:1.Institute
It is optimum proportioning to match ratio, can either efficiently digest oyster, will not cause the waste of raw material.
The present invention adds suitable ficin, takes suitable temperature and time in oyster shells step, can
It is efficiently quick to reach mechanical dejacketing, save the time.The amount and enzymatic hydrolysis condition used when enzymolysis all just reaches just
The degree shelled, does not lose Oyster Protein.
Compared with prior art, the beneficial effects of the present invention are：
1st, the present invention is shelled oyster using enzymatic isolation method, is reached mechanical dejacketing, is saved the time, shortens the production cycle, can
Realize large-scale industrialized production.
2nd, the present invention is carried out at the same time homogenization in enzymolysis process, greatly improves hydrolysis result, enzymolysis time shortens
More than one times, while when to find homogenization pressure be 16~21Mpa, it is possible to achieve the quality of product and production cost it is perfectly balanced.
3rd, the present invention uses complex enzyme zymohydrolysis, finds best of breed point and enzymatic hydrolysis condition, the small-molecular peptides digested point
Son amount is small, and activity is high.
Embodiment
The invention will be further described with reference to embodiments, but the invention is not limited in these embodiments.
Embodiment 1
(1) oyster is cleaned, adds oyster 7 times of water of weight, add the ficin of oyster weight 0.03%, adjust
PH value is saved to 4.7,54 DEG C is warming up to, digests 20min, take oyster meat；
(2) oyster meat is smashed, adds 4 times of water of its weight, stir evenly, obtain oyster slurries；
(3) complex enzyme of oyster meat weight 0.09% is added, adjusts pH to 6.5, rise temperature is in pressure to 46 DEG C
Under 16Mpa, homogeneous 140min, temperature is improved to 75 DEG C of progress enzyme deactivations, obtains enzymolysis liquid, the complex enzyme is that weight ratio is 4:
2:The mixture of 1 ficin, pronase and subtilopeptidase A；
(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.
Embodiment 2
(1) oyster is cleaned, adds oyster 8 times of water of weight, add the ficin of oyster weight 0.04%, adjust
PH value is saved to 4.9,52 DEG C is warming up to, digests 18min, take oyster meat；
(2) oyster meat is smashed, adds 3 times of water of its weight, stir evenly, obtain oyster slurries；
(3) complex enzyme of oyster meat weight 0.08% is added, adjusts pH to 5.3, rise temperature is in pressure to 47 DEG C
Under 17Mpa, homogeneous 130min, temperature is improved to 76 DEG C of progress enzyme deactivations, obtains enzymolysis liquid, the complex enzyme is that weight ratio is 4:
2:The mixture of 1 ficin, pronase and subtilopeptidase A；
(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.
Embodiment 3
(1) oyster is cleaned, adds oyster 9 times of water of weight, add the ficin of oyster weight 0.05%, adjust
PH value is saved to 4.8,53 DEG C is warming up to, digests 15min, take oyster meat；
(2) oyster meat is smashed, adds 3 times of water of its weight, stir evenly, obtain oyster slurries；
(3) complex enzyme of oyster meat weight 0.07% is added, adjusts pH to 5.9, rise temperature is in pressure to 49 DEG C
Under 18Mpa, homogeneous 120min, temperature is improved to 77 DEG C of progress enzyme deactivations, obtains enzymolysis liquid, the complex enzyme is that weight ratio is 4:
2:The mixture of 1 ficin, pronase and subtilopeptidase A；
(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.
Embodiment 4
(1) oyster is cleaned, adds oyster 10 times of water of weight, add the ficin of oyster weight 0.06%,
PH value is adjusted to 48,51 DEG C is warming up to, digests 13min, take oyster meat；
(2) oyster meat is smashed, adds 2 times of water of its weight, stir evenly, obtain oyster slurries；
(3) complex enzyme of oyster meat weight 0.05% is added, adjusts pH to 5.5, rise temperature is in pressure to 50 DEG C
Under 19Mpa, homogeneous 110min, temperature is improved to 78 DEG C of progress enzyme deactivations, obtains enzymolysis liquid, the complex enzyme is that weight ratio is 4:
2:The mixture of 1 ficin, pronase and subtilopeptidase A；
(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.
Embodiment 5
(1) oyster is cleaned, adds oyster 10 times of water of weight, add the ficin of oyster weight 0.08%,
PH value is adjusted to 4.9,52 DEG C is warming up to, digests 10min, take oyster meat；
(2) oyster meat is smashed, adds 3 times of water of its weight, stir evenly, obtain oyster slurries；
(3) complex enzyme of oyster meat weight 0.04% is added, adjusts pH to 6.2, rise temperature is in pressure to 52 DEG C
Under 21Mpa, homogeneous 100min, temperature is improved to 80 DEG C of progress enzyme deactivations, obtains enzymolysis liquid, the complex enzyme is that weight ratio is 4:
2:The mixture of 1 ficin, pronase and subtilopeptidase A；
(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides.
In the product that the present invention obtains, more than 90%, the molecular weight of 95% oyster small-molecular peptides exists protein content
Below 480Dalton.Following table obtains oyster small-molecular peptides molecular weight distribution table for the present invention：
The present invention carries out homogeneous operation under the conditions of pressure is 16~21Mpa, is that inventor passes through substantial amounts of comparative experiments
It was found that the quality of product is not proportional with homogenization pressure, part correction data is now listed：
Claims (2)
- A kind of 1. method that small-molecular peptides are prepared with complex enzyme for hydrolyzing Oyster Protein, it is characterised in that including following preparation process：(1) oyster is cleaned, adds oyster 7~10 times of water of weight, add the fig egg of oyster weight 0.03~0.08% White enzyme, adjusts pH value to 4.7~4.9, is warming up to 51~54 DEG C, digests 10~20min, take oyster meat；(2) oyster meat is smashed, adds 2~4 times of water of its weight, stir evenly, obtain oyster slurries；(3) complex enzyme of oyster meat weight 0.04~0.09% is added, adjusts pH to 5.3~6.5, rise temperature to 46~52 DEG C, 100~140min of homogeneous, enzyme deactivation, obtains enzymolysis liquid；(4) enzymolysis liquid is subjected to ultrafiltration, is freeze-dried, obtains oyster small-molecular peptides；The complex enzyme is that weight ratio is 4:2:1 ficin, pronase and subtilopeptidase A it is mixed Compound；Homogenization pressure is 16~21Mpa in the step (3).
- 2. the method according to claim 1 that small-molecular peptides are prepared with complex enzyme for hydrolyzing Oyster Protein, it is characterised in that：Step Suddenly the temperature of enzyme deactivation is improved to 75~80 DEG C in (3).
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Families Citing this family (6)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN105326035B (en) * | 2015-10-19 | 2018-12-04 | 集美大学 | A kind of production method of less salt oyster polypeptide and oligosaccharide nutrient powder |
| CN106107635A (en) * | 2016-06-29 | 2016-11-16 | 大连深蓝肽科技研发有限公司 | Utilize the method that Concha Ostreae fresh meat prepares Concha Ostreae oligopeptide |
| CN106243187B (en) * | 2016-09-05 | 2019-08-20 | 华南理工大学 | A method of the quick separating oyster peptide from oyster enzymolysis liquid |
| CN110819677B (en) * | 2019-11-29 | 2021-08-17 | 烟台大学 | Preparation method of debitterized antioxidant oyster peptide |
| CN113215216A (en) * | 2021-05-28 | 2021-08-06 | 江苏格局生物医药科技有限公司 | Industrial production method for preparing oyster peptide by enzyme method |
| CN114350733B (en) * | 2021-12-29 | 2024-01-26 | 广东冠龙生物科技有限公司 | Oyster active protein peptide composite accurate enzymolysis technology |
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