CN109369775A - Low purine marine fishes oligopeptide and preparation method thereof - Google Patents
Low purine marine fishes oligopeptide and preparation method thereof Download PDFInfo
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- CN109369775A CN109369775A CN201811283736.9A CN201811283736A CN109369775A CN 109369775 A CN109369775 A CN 109369775A CN 201811283736 A CN201811283736 A CN 201811283736A CN 109369775 A CN109369775 A CN 109369775A
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
- C07K1/306—Extraction; Separation; Purification by precipitation by crystallization
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
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- 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
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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Abstract
The present invention relates to a kind of low purine marine fishes oligopeptides and preparation method thereof.The preparation method includes the following steps: that (1) obtains marine fishes enzymolysis liquid, and adjusts the pH to more than 7 of the marine fishes enzymolysis liquid；Then fresh milk is added to be reacted；(2) pH to less than 7 of reaction solution obtained by regulating step (1) and holding extremely crystallization, obtain pretreatment fluid；(3) pretreatment fluid is first subjected to ceramic filter, then carries out ultrafiltration, collect filtrate, be concentrated and dried to get.The preparation method more can completely remove the combination purine in free state purine and nucleic acid molecule, and the purine content of marine fishes oligopeptide is effectively reduced.
Description
Technical field
The present invention relates to marine fishes productions, more particularly to low purine marine fishes oligopeptide and its preparation side
Method.
Background technique
Ocean fish is resourceful, and nutritive value is high, and delicious flavour is the precious good merchantable brand on dining table.Marine fishes albumen is through enzyme
The marine fishes oligopeptide of method hydrolysis preparation, not only has nutritive value high, be absorbed and utilized it is fast, it is soluble easily in water, convenient for
A variety of advantages such as other food compositions are compatible, and there are the multiple biological activities such as anti-oxidant, antibacterial, blood pressure lowering, antitumor,
It is good functional food ingredient, but because purine content is high, is not suitable for the food ingredient as gout patients, in order to improve pain
How the life quality of wind patient reduces the purine content of marine fishes oligopeptide, becomes a good problem to study.
Currently, main method has boiling, saltouts, adsorbs in domestic removing food in the scheme of purine.Wherein, boiling
Method is suitble to solid food, can remove free state purine, cannot remove the combination purine in nucleic acid molecule, while food is water-soluble
Component damages are serious；Absorption and salting out method can only remove free state purine in liquid food, and industrialization is at high cost, big for nucleic acid
Combination purine in molecule can not be removed directly.
Summary of the invention
Based on this, it is necessary to provide a kind of preparation method of low purine marine fishes oligopeptide, which can be compared with
Completely to remove the combination purine in free state purine and nucleic acid molecule, the purine that marine fishes oligopeptide is effectively reduced contains
Amount.
A kind of preparation method of low purine marine fishes oligopeptide, includes the following steps:
(1) marine fishes enzymolysis liquid is obtained, and adjusts the pH to more than 7 of the marine fishes enzymolysis liquid；Then it is added new
Fresh cow milk is reacted；
(2) pH to less than 7 of reaction solution obtained by regulating step (1) and holding extremely crystallization, obtain pretreatment fluid；
(3) pretreatment fluid is first subjected to ceramic membrane filter, then carries out ultrafiltration, collect filtrate, be concentrated and dried to get.
In one of the embodiments, in step (1), the condition of the reaction are as follows: -55 DEG C of temperature 45 C, time 1.5-
3.5h.Preferably 52-55 DEG C of temperature, time 2-2.5h.
In one of the embodiments, in step (1), the volume ratio of the fresh milk and marine fishes enzymolysis liquid is
0.5:100-2:100.Preferably 1:100-1.5:100.
In one of the embodiments, in step (1), the pH to 7.5-8.5 of the marine fishes enzymolysis liquid is adjusted.
In one of the embodiments, in step (1), the pH of the marine fishes enzymolysis liquid is adjusted using sodium hydroxide.
In one of the embodiments, in step (2), the pH to 4-6 of reaction solution obtained by regulating step (1).
In one of the embodiments, in step (2), using the pH of reaction solution obtained by citric acid regulating step (1).
In one of the embodiments, in step (2), the time of the holding to crystallization is 1.5-3h.
In one of the embodiments, in step (3), the ceramic membrane that the ceramic membrane filter uses is made pottery for 0.1-0.3 μm
Porcelain film.
The molecular cut off for the ultrafiltration membrane that the ultrafiltration uses in one of the embodiments, is 4000-6000u.
In one of the embodiments, in step (3), the concentration is vacuum concentration.
In one of the embodiments, in step (3), the method for the drying is spray drying.
The method for obtaining the marine fishes enzymolysis liquid in one of the embodiments, is as follows:
Marine fish after pre-treatment is blended, water and regulation system pH to 7-8 is added, animal proteolytic enzyme and wind is added
Taste enzyme is digested；After the enzymatic hydrolysis, enzyme deactivation is then centrifuged for removing residue and grease.
The pre-treatment includes decaptitating, removes internal organ, remove the gill, scale in one of the embodiments,.
The dosage of the animal proteolytic enzyme is the 0.4- of the marine fish weight in one of the embodiments,
0.6%；The dosage of the food flavor enzyme is the 0.08-0.12% of the marine fish weight.
In one of the embodiments, in step (1), the temperature of the enzymatic hydrolysis is 45-55 DEG C, time 2-4h.
In one of the embodiments, in step (1), the temperature of the enzyme deactivation is 90-95 DEG C, time 10-20min.
The filter cloth used that is centrifuged is 500-700 mesh filter cloth in one of the embodiments,.
The present invention also provides the low purine marine fishes oligopeptides that the preparation method is prepared.
Compared with prior art, the invention has the following beneficial effects:
The preparation method of marine fishes oligopeptide of the invention, using fresh milk by marine fishes under certain pH conditions
Free purine in enzymolysis liquid is converted into uric acid, then induces uric acid to form crystallization by adjusting pH value, in conjunction with ceramic filter
With the characteristic of ultra-filtration and separation, while filtering out uric acid crystal, additionally it is possible to effectively removing macromolecular nucleic acid in combine purine, remove it is fast
Purine rate is high, thus greatly reduces the purine content in the marine fishes oligopeptide being prepared, can be used as gout patients
Quality-assured food ingredient, and it is full of nutrition, safe and non-toxic.Meanwhile the preparation method technical process is simple, operation is easy, is at low cost.
Specific embodiment
Low purine marine fishes oligopeptide of the invention and preparation method thereof is made below in conjunction with specific embodiment further
Detailed description.
The marine fishes enzymolysis liquid used in 1-5 of the embodiment of the present invention and comparative example 1-2 is made by the steps to obtain:
It by marine fishes (such as stripped tuna, Keeping of Pilchard) decaptitating, skin, bone and internal organ, is rubbed after cleaning, by material water quality
Amount adds water to be homogenized than 1:1, and regulation system pH to 7.5 adds 0.5% animal proteolytic enzyme of marine fish weight and 0.1%
Food flavor enzyme, digests 3h under the conditions of temperature 50 C, pH7.5, is then warming up to 90~95 DEG C of enzyme deactivation 10min, then be centrifuged with tripodia
Grease and residue are removed in machine (built-in 600 mesh cloth bag) 3000r/min centrifugation, and marine fishes enzymolysis liquid is made.
Embodiment 1
A kind of preparation method of low purine marine fishes oligopeptide of the present embodiment, the raw material of use is with embodiment 1, and step is such as
Under:
(1) by marine fishes enzymolysis liquid 1.0M sodium hydroxide solution tune pH to 8.0, fresh milk is pressed into 0.5:100 body
Product ratio is added in marine fishes enzymolysis liquid, reacts 3.5h at 50 DEG C；
(2) after reaction, with the pH to 4.0 of 1.0M citric acid tune reaction solution, 2.0h is kept, crystallization obtains pretreatment fluid；
(3) by 0.2 μm of ceramic membrane filter of pretreatment fluid, ultrafiltration then is carried out with the ultrafiltration membrane of molecular cut off 5000u；
Marine fishes oligopeptide is made through vacuum concentration and spray drying in the ultrafiltration permeate being collected into, and removes purine rate 88%.
Embodiment 2
A kind of preparation method of low purine marine fishes oligopeptide of the present embodiment, the raw material of use is with embodiment 1, and step is such as
Under:
(1) by marine fishes enzymolysis liquid 1.0M sodium hydroxide solution tune pH to 8.5, fresh milk is pressed into 1.0:100 body
Product ratio is added in marine fishes enzymolysis liquid, reacts 3.0h at 50 DEG C；
(2) after reaction, with the pH to 4.5 of 1.0M citric acid tune reaction solution, 2.5h is kept, crystallization obtains pretreatment fluid；
(3) by 0.2 μm of ceramic membrane filter of pretreatment fluid, ultrafiltration then is carried out with the ultrafiltration membrane of molecular cut off 5000u；
Low purine marine fishes oligopeptide is made through vacuum concentration and spray drying in the ultrafiltration permeate being collected into, and removes purine rate
93%.
Embodiment 3
A kind of preparation method of low purine marine fishes oligopeptide of the present embodiment, the raw material of use is with embodiment 1, and step is such as
Under:
(1) by marine fishes enzymolysis liquid 1.0M sodium hydroxide solution tune pH to 8.0, fresh milk is pressed into 1.5:100 body
Product ratio is added in marine fishes enzymolysis liquid, reacts 2.5h at 55 DEG C；
(2) after reaction, with the pH to 4.5 of 1.0M citric acid tune reaction solution, 2.0h is kept, crystallization obtains pretreatment fluid；
(3) by 0.2 μm of ceramic membrane filter of pretreatment fluid, ultrafiltration then is carried out with the ultrafiltration membrane of molecular cut off 5000u；
Marine fishes oligopeptide is made through vacuum concentration and spray drying in the ultrafiltration permeate being collected into, and removes purine rate 96%.
Embodiment 4
A kind of preparation method of marine fishes oligopeptide of the present embodiment, the raw material of use is with embodiment 1, and steps are as follows:
(1) by marine fishes enzymolysis liquid 1.0M sodium hydroxide solution tune pH to 8.0, fresh milk is pressed into 2:100 volume
Than being added in marine fishes enzymolysis liquid, 1.5h is reacted at 50 DEG C；
(2) after reaction, with the pH to 4.0 of 1.0M citric acid tune reaction solution, 1.5h is kept, crystallization obtains pretreatment fluid；
(3) by 0.2 μm of ceramic membrane filter of pretreatment fluid, ultrafiltration then is carried out with the ultrafiltration membrane of molecular cut off 5000u；
Marine fishes oligopeptide is made through vacuum concentration and spray drying in the ultrafiltration permeate being collected into, and removes purine rate 92%.
Embodiment 5
A kind of preparation method of marine fishes oligopeptide of the present embodiment, the raw material of use is with embodiment 1, and steps are as follows:
(1) by marine fishes enzymolysis liquid 1.0M sodium hydroxide solution tune pH to 7.5, fresh milk is pressed into 1.5:100 body
Product ratio is added in marine fishes enzymolysis liquid, reacts 3h at 50 DEG C；
(2) after reaction, with the pH to 4.0 of 1.0M citric acid tune reaction solution, 3h is kept, crystallization obtains pretreatment fluid；
(3) by 0.2 μm of ceramic membrane filter of pretreatment fluid, ultrafiltration then is carried out with the ultrafiltration membrane of molecular cut off 5000u；
Marine fishes oligopeptide is made through vacuum concentration and spray drying in the ultrafiltration permeate being collected into, and removes purine rate 90%.
Comparative example 1
A kind of preparation method of marine fishes oligopeptide of this comparative example, with embodiment 1, difference exists the raw material and step of use
In: without the ceramic membrane filter in step (3).It is 60% that the preparation method, which removes purine rate,.
Comparative example 2
A kind of preparation method of marine fishes oligopeptide of this comparative example, with embodiment 1, difference exists the raw material and step of use
In: the ceramic membrane filter is substituted using macroreticular resin AB-8 absorption in step (3).The preparation method removes purine rate
72%.
Each technical characteristic of embodiment described above can be combined arbitrarily, for simplicity of description, not to above-mentioned reality
It applies all possible combination of each technical characteristic in example to be all described, as long as however, the combination of these technical characteristics is not deposited
In contradiction, all should be considered as described in this specification.
The embodiments described above only express several embodiments of the present invention, and the description thereof is more specific and detailed, but simultaneously
It cannot therefore be construed as limiting the scope of the patent.It should be pointed out that coming for those of ordinary skill in the art
It says, without departing from the inventive concept of the premise, various modifications and improvements can be made, these belong to protection of the invention
Range.Therefore, the scope of protection of the patent of the invention shall be subject to the appended claims.
Claims (10)
1. a kind of preparation method of low purine marine fishes oligopeptide, which comprises the steps of:
(1) marine fishes enzymolysis liquid is obtained, and adjusts the pH to more than 7 of the marine fishes enzymolysis liquid；Then fresh ox is added
Cream is reacted；
(2) pH to less than 7 of reaction solution obtained by regulating step (1) and holding extremely crystallization, obtain pretreatment fluid；
(3) pretreatment fluid is first subjected to ceramic membrane filter, then carries out ultrafiltration, collect filtrate, be concentrated and dried to get.
2. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (1),
The condition of the reaction are as follows: -55 DEG C of temperature 45 C, time 1.5-3.5h.
3. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (1),
The volume ratio of the fresh milk and marine fishes enzymolysis liquid is 0.5:100-2:100.
4. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (1),
Adjust the pH to 7.5-8.5 of the marine fishes enzymolysis liquid.
5. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (2),
The pH to 4-6 of reaction solution obtained by regulating step (1).
6. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (2),
The time of the holding to crystallization is 1.5-3h.
7. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (3),
The ceramic membrane that the ceramic membrane filter uses is 0.1-0.3 μm of ceramic membrane.
8. the preparation method of low purine marine fishes oligopeptide according to claim 1, which is characterized in that in step (3),
The molecular cut off for the ultrafiltration membrane that the ultrafiltration uses is 4000-6000u.
9. the preparation method of low purine marine fishes oligopeptide according to claim 1-8, which is characterized in that obtain
The method for taking the marine fishes enzymolysis liquid is as follows:
Marine fish after pre-treatment is blended, water and regulation system pH to 7-8 is added, animal proteolytic enzyme and food flavor enzyme is added
It is digested；After the enzymatic hydrolysis, enzyme deactivation is then centrifuged for removing residue and grease.
10. the low purine marine fishes oligopeptide that the described in any item preparation methods of claim 1-9 are prepared.
Priority Applications (2)
| Application Number | Priority Date | Filing Date | Title |
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| CN201811283736.9A CN109369775B (en) | 2018-10-31 | 2018-10-31 | Low-purine marine fish oligopeptide and preparation method thereof |
| PCT/CN2019/080660 WO2020087857A1 (en) | 2018-10-31 | 2019-03-29 | Low-purine marine fish oligopeptide and preparation method therefor |
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| CN201811283736.9A CN109369775B (en) | 2018-10-31 | 2018-10-31 | Low-purine marine fish oligopeptide and preparation method thereof |
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Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
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| WO2020087857A1 (en) * | 2018-10-31 | 2020-05-07 | 广东兴亿海洋生物工程股份有限公司 | Low-purine marine fish oligopeptide and preparation method therefor |
| CN112592316A (en) * | 2020-12-11 | 2021-04-02 | 山东瑞芝生物科技股份有限公司 | Method for reducing purine content in tuna anserine raw material by using membrane filtration and application thereof |
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| CN112592316A (en) * | 2020-12-11 | 2021-04-02 | 山东瑞芝生物科技股份有限公司 | Method for reducing purine content in tuna anserine raw material by using membrane filtration and application thereof |
| CN112592316B (en) * | 2020-12-11 | 2021-11-23 | 山东瑞芝生物科技股份有限公司 | Method for reducing purine content in tuna anserine raw material by using membrane filtration and application thereof |
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| CN109369775B (en) | 2020-12-08 |
| WO2020087857A1 (en) | 2020-05-07 |
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