CN108157580A - A kind of pork functional polypeptide and preparation method thereof - Google Patents
A kind of pork functional polypeptide and preparation method thereof Download PDFInfo
- Publication number
- CN108157580A CN108157580A CN201711482472.5A CN201711482472A CN108157580A CN 108157580 A CN108157580 A CN 108157580A CN 201711482472 A CN201711482472 A CN 201711482472A CN 108157580 A CN108157580 A CN 108157580A
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- China
- Prior art keywords
- pork
- preparation
- enzymolysis
- functional polypeptide
- polypeptide
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Classifications
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- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J1/00—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites
- A23J1/02—Obtaining protein compositions for foodstuffs; Bulk opening of eggs and separation of yolks from whites from meat
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23J—PROTEIN COMPOSITIONS FOR FOODSTUFFS; WORKING-UP PROTEINS FOR FOODSTUFFS; PHOSPHATIDE COMPOSITIONS FOR FOODSTUFFS
- A23J3/00—Working-up of proteins for foodstuffs
- A23J3/30—Working-up of proteins for foodstuffs by hydrolysis
- A23J3/32—Working-up of proteins for foodstuffs by hydrolysis using chemical agents
- A23J3/34—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes
- A23J3/341—Working-up of proteins for foodstuffs by hydrolysis using chemical agents using enzymes of animal proteins
Abstract
The invention discloses the preparation method of pork functional polypeptide, the pretreatment including pork, the preparation of the enzymolysis and polypeptide of pork.Invention also discloses the pork functional polypeptides prepared by this method.It has the beneficial effect that：Preparation method of the present invention is quick and easy, and the combination degree of inhibitor contained in substrate and enzyme is small, and enzyme digestion reaction rate is fast, substantially reduces the time of enzymolysis, while enzyme digestion reaction is abundant, and pork resource is made to be fully used；Prepared polypeptide can remove free radical, anti-lipid peroxidation and protect cells from oxidative damage.
Description
Technical field
The present invention relates to biologically active peptide field, more particularly, to a kind of pork functional polypeptide and preparation method thereof.
Background technology
Biologically active peptide is amino acid includes the annular, linear of dipeptides and complexity with what different arrangement modes and combination were formed
The general name of the different peptides of structure.Different biological peptides has different structure and physiological function, such as antiviral, anticancer, anti-blood
The effects that bolt, anti-hypertension, immunological regulation, hormone control, antibacterial, norcholesterol.Pork is the main non-staple food of daily life,
With qi-restoratives salubrity, nourishing Yin and moistening dryness, riching and moisturizing skim effect.At this stage, it is less for the research of pork polypeptide.
Invention content
The purpose of the present invention is to provide the preparation method of pork functional polypeptide, the polypeptide preparation efficiency is high, during enzymolysis
Between it is few, and digest abundant, pork resource made to be fully used.
Another object of the present invention is to provide a kind of pork functional polypeptide, which can remove free radical, inhibit
Lipid peroxidation and protect cells from oxidative damage.
The technical solution taken to achieve the above object of the present invention is：The preparation method of pork functional polypeptide, this is more
The preparation method of peptide includes the following steps：
Pork pre-processes：Fresh pork of slaughtering is freezed into 25~30h at temperature -25~-30 DEG C, which can extend
The shelf-life of pork provides the raw material of high-quality for enzymolysis process；
The enzymolysis of pork：Frozen Pork is taken to thaw, distilled water, solid-liquid ratio 1g are added in into the pork of defrosting:3~7mL adjusts pH
It is 6.5~7.5, adds in the papain of pork quality 1.0%~1.5%, digests 4~6h at 50~55 DEG C, be heated to 90 DEG C
Enzyme deactivation live 10min, adjust pH be 9~10, add in pork quality 0.8%~1.2% alkali protease, 42~50 DEG C digest 3~
6h, is heated to 90 DEG C of enzyme deactivations work 10min, is cooled to room temperature, and 10000r/min centrifugation 15min collect supernatant, obtain pork enzymolysis
Liquid, under this condition, inhibitor contained in substrate and the combination degree of enzyme are small, enzyme digestion reaction rate is fast, substantially reduces enzymolysis
Time, while enzyme digestion reaction is abundant, and pork resource is made to be fully used；
The preparation of polypeptide：Pork enzymatic hydrolyzate through the ultrafiltration membrane that molecular cut off is 1kDa is filtered, trapped fluid is collected, will cut
Liquid is stayed to be added in the chromatographic column equipped with 8~10 times of D101 macroreticular resins according to volume ratio, with 3~5 times of column volume washing removings
Then decontamination is eluted with the mass fraction of 5~8 times of column volumes for 60%~62% ethanol water, ethanol eluate exists
50 DEG C of revolvings remove ethyl alcohol, are freeze-dried to obtain polypeptide.The process is easy to operate, reproducible, and Elution range is wide, can be according to reality
Border situation is amplified condition and realizes industrialized production.
Compared with prior art, beneficial effects of the present invention are：The present invention by the pretreatment of pork, enzymolysis and etc. system
Pork functional polypeptide is obtained, preparation method is simple and quick, and the combination degree of inhibitor contained in substrate and enzyme is small, and enzymolysis is anti-
It answers rate fast, substantially reduces the time of enzymolysis, while enzyme digestion reaction is abundant, pork resource is made to be fully used；It is made
Standby polypeptide can remove free radical, anti-lipid peroxidation and protect cells from oxidative damage.
Specific embodiment
It is described in further detail with reference to embodiments：
Embodiment 1：The preparation method of pork functional polypeptide, includes the following steps：
1) pork pre-processes：It slaughters pork by fresh at -25 DEG C of temperature and freezes 25h, which can extend pork
Shelf-life provides the raw material of high-quality for enzymolysis process；
2) enzymolysis of pork：Frozen Pork is taken to thaw, distilled water, solid-liquid ratio 1g are added in into the pork of defrosting:3mL adjusts pH
It is 6.5, adds in the papain of pork quality 1.0%, 4h is digested at 50 DEG C, being heated to 90 DEG C of enzyme deactivations work 10min, tune pH is
9, the alkali protease of pork quality 0.8% is added in, 3h is digested at 42 DEG C, 90 DEG C of enzyme deactivations work 10min is heated to, is cooled to room temperature,
10000r/min centrifuges 15min, collects supernatant, obtains pork enzymatic hydrolyzate, under this condition, the inhibitor and enzyme contained by substrate
Combination degree is small, and enzyme digestion reaction rate is fast, substantially reduces the time of enzymolysis, while enzyme digestion reaction is abundant, obtains pork resource
To making full use of；
3) preparation of polypeptide：Pork enzymatic hydrolyzate through the ultrafiltration membrane that molecular cut off is 1kDa is filtered, collects trapped fluid, it will
Trapped fluid is added to according to volume ratio in the chromatographic column equipped with 8 times of D101 macroreticular resins, and impurity elimination is removed with 3 times of column volume water elutions
Then matter is eluted with the mass fraction of 5 times of column volumes for 60% ethanol water, ethanol eluate is removed in 50 DEG C of revolvings
Ethyl alcohol is freeze-dried to obtain polypeptide.The process is easy to operate, reproducible, and Elution range is wide, can be put according to actual conditions
Big condition realizes industrialized production.
Embodiment 2：The preparation method of pork functional polypeptide, includes the following steps：It is pork pretreatment first：It will be fresh
It slaughters pork and freezes 30h at -30 DEG C of temperature；Followed by the enzymolysis of pork：Frozen Pork is taken to thaw, is added into the pork of defrosting
Enter distilled water, solid-liquid ratio 1g:7mL, it is 7.5 to adjust pH, adds in the papain of pork quality 1.5%, and 6h is digested at 55 DEG C,
90 DEG C of enzyme deactivations work 10min are heated to, it is 10 to adjust pH, adds in the alkali protease of pork quality 1.2%, digests 6h at 50 DEG C, heats
It to 90 DEG C of enzyme deactivations work 10min, is cooled to room temperature, 10000r/min centrifugation 15min collect supernatant, obtain pork enzymatic hydrolyzate；Finally
It is the preparation of polypeptide：Pork enzymatic hydrolyzate through the ultrafiltration membrane that molecular cut off is 1kDa is filtered, trapped fluid is collected, will retain
Liquid is added to according to volume ratio in the chromatographic column equipped with 10 times of D101 macroreticular resins, removes impurity with 5 times of column volume water elutions, so
Eluted afterwards with the mass fraction of 8 times of column volumes for 62% ethanol water, ethanol eluate 50 DEG C of revolvings remove ethyl alcohol,
It is freeze-dried to obtain polypeptide.The process is easy to operate, reproducible, and Elution range is wide, can be amplified item according to actual conditions
Part realizes industrialized production.
Embodiment 3：The preparation method of pork functional polypeptide, includes the following steps：
1) pork digests：Frozen Pork is taken to thaw, distilled water, solid-liquid ratio 1g are added in into the pork of defrosting:7mL, tune pH are
7.5, the papain of pork quality 1.5% is added in, adds 3, the 5- resacetophenones and pork of pork quality 0.01%
The aziridine of quality 0.008% digests 2h at 55 DEG C, is heated to 90 DEG C of enzyme deactivations work 10min, it is 10 to adjust pH, adds in Meat
The alkali protease of amount 1.2% digests 6h at 50 DEG C, is heated to 90 DEG C of enzyme deactivations and lives 10min, is cooled to room temperature, 10000r/min from
Heart 15min collects supernatant, obtains pork enzymatic hydrolyzate；3,5- resacetophenones, aziridine and the papain added in
In amino acid residue His-159 have synergistic effect, be greatly promoted the deprotonation of Cys-25, Cys-25 made to be more easy to attack
The carbonyl carbon in peptide backbone is hit, significantly provides enzymolysis efficiency, shortens the manufacturing cycle of pork functional polypeptide.
2) preparation of polypeptide：Pork enzymatic hydrolyzate through the ultrafiltration membrane that molecular cut off is 1kDa is filtered, collects retention
Trapped fluid is added to according to volume ratio in the chromatographic column equipped with 10 times of D101 macroreticular resins by liquid, with 5 times of column volume washing removings
Then decontamination is eluted with the mass fraction of 8 times of column volumes for 62% ethanol water, ethanol eluate is in 50 DEG C of revolvings
It removes ethyl alcohol, be freeze-dried to obtain polypeptide.
Embodiment of above is merely to illustrate the present invention, and not limitation of the present invention, the ordinary skill people of this field
Member without departing from the spirit and scope of the present invention, can also make a variety of changes and modification.Therefore, it is all equivalent
Technical solution also belong to scope of the invention, scope of patent protection of the invention should be defined by the claims.
Claims (8)
1. the preparation method of pork functional polypeptide, the pretreatment including pork, the enzymolysis of pork, the preparation of polypeptide,
It is characterized in that：The enzymolysis process of the pork is：Frozen Pork is taken to thaw, distilled water is added in into the pork of defrosting,
Solid-liquid ratio is 1g:3~7mL, it is 6.5~7.5 to adjust pH, adds in papain, enzymolysis, and enzyme deactivation is lived, and it is 9~10 to adjust pH, is added in
Alkali protease, enzymolysis, enzyme deactivation are lived, are cooled to room temperature, and 10000r/min centrifugation 15min collect supernatant, obtain pork enzymolysis
Liquid.
2. the preparation method of pork functional polypeptide according to claim 1, it is characterised in that：The papain
Addition is the 1.0%~1.5% of pork weight, and hydrolysis temperature is 50~55 DEG C, and enzymolysis time is 4~6h.
3. the preparation method of pork functional polypeptide according to claim 1, it is characterised in that：The alkali protease
Addition is the 0.8%~1.2% of pork weight, and hydrolysis temperature is 42~50 DEG C, and enzymolysis time is 3~6h.
4. the preparation method of pork functional polypeptide according to claim 1, it is characterised in that：The pork is pretreated
Cheng Wei：Fresh pork of slaughtering is freezed into 25~30h at temperature -25~-30 DEG C.
5. the preparation method of pork functional polypeptide according to claim 1, it is characterised in that：The preparation of the polypeptide
Cheng Wei：Pork enzymatic hydrolyzate through ultrafiltration membrane is filtered, collects trapped fluid, trapped fluid is added to equipped with 8 according to volume ratio~
In the chromatographic column of 10 times of D101 macroreticular resins, impurity are removed with 3~5 times of column volume water elutions, then with 5~8 times of column volumes
Ethanol water is eluted, and revolving removes ethyl alcohol, dry polypeptide.
6. the preparation method of pork functional polypeptide according to claim 5, it is characterised in that：The retention of the ultrafiltration membrane
Molecular weight is 1kDa.
7. the preparation method of pork functional polypeptide according to claim 5, it is characterised in that：In the ethanol water
The mass fraction of ethyl alcohol is 60%~62%.
8. pork functional polypeptide prepared by any one of claim 1-7 the methods.
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711482472.5A CN108157580A (en) | 2017-12-29 | 2017-12-29 | A kind of pork functional polypeptide and preparation method thereof |
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| Application Number | Priority Date | Filing Date | Title |
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| CN201711482472.5A CN108157580A (en) | 2017-12-29 | 2017-12-29 | A kind of pork functional polypeptide and preparation method thereof |
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| CN108157580A true CN108157580A (en) | 2018-06-15 |
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110714046A (en) * | 2019-11-22 | 2020-01-21 | 浙江丰安生物制药有限公司 | Preparation method of polypeptide for hydrolyzing pork protein of suckling pig |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1970784A (en) * | 2006-12-07 | 2007-05-30 | 伍曾利 | Method for extracting carnosine |
| CN105648010A (en) * | 2016-02-23 | 2016-06-08 | 浙江海洋学院 | Preparation method of Sphyrna lewini meat antioxidative peptides capable of activating Nrf2-ARE pathway |
| CN107279451A (en) * | 2017-05-11 | 2017-10-24 | 浙江海洋大学 | The preparation method of hairtail antioxidation activity peptidase hydrolyzed liquor |
-
2017
- 2017-12-29 CN CN201711482472.5A patent/CN108157580A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1970784A (en) * | 2006-12-07 | 2007-05-30 | 伍曾利 | Method for extracting carnosine |
| CN105648010A (en) * | 2016-02-23 | 2016-06-08 | 浙江海洋学院 | Preparation method of Sphyrna lewini meat antioxidative peptides capable of activating Nrf2-ARE pathway |
| CN107279451A (en) * | 2017-05-11 | 2017-10-24 | 浙江海洋大学 | The preparation method of hairtail antioxidation activity peptidase hydrolyzed liquor |
Non-Patent Citations (1)
| Title |
|---|
| 都荣强等: "猪肉不同蛋白酶解呈味组分及热反应风味物质比较", 《中国食品学报》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN110714046A (en) * | 2019-11-22 | 2020-01-21 | 浙江丰安生物制药有限公司 | Preparation method of polypeptide for hydrolyzing pork protein of suckling pig |
| CN110714046B (en) * | 2019-11-22 | 2021-06-04 | 浙江丰安生物制药有限公司 | Preparation method of polypeptide for hydrolyzing pork protein of suckling pig |
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Application publication date: 20180615 |