CN107460224A - A kind of preparation method of samara oligopeptide - Google Patents
A kind of preparation method of samara oligopeptide Download PDFInfo
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- CN107460224A CN107460224A CN201710901028.6A CN201710901028A CN107460224A CN 107460224 A CN107460224 A CN 107460224A CN 201710901028 A CN201710901028 A CN 201710901028A CN 107460224 A CN107460224 A CN 107460224A
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- samara
- oligopeptide
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- C—CHEMISTRY; METALLURGY
- 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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- 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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Abstract
The present invention provides a kind of preparation method of samara oligopeptide, and it with distilled water using samara crude protein as raw material, to mix, with homogenizer homogeneous, temperature control adjusts pH, adds protease, enzymolysis, enzyme deactivation, in being centrifuged on centrifuge, supernatant is taken, solid formation is dissolved in ethanol after centrifugation, homogenizer homogeneous, temperature control adjusts pH, adds protease, enzymolysis, enzyme deactivation, in being centrifuged on centrifuge, take supernatant, merge supernatant, over-molecular sieve, dry samara oligopeptide dry powder.Digested in the present invention using compound protease, and samara oligopeptide is prepared from two kinds of solvents of pure water and ethanol solution, effectively increase the degree of hydrolysis of protein, by the peptide that the samara protein dissolution of macromolecular is small molecule.
Description
Technical field
The present invention relates to a kind of method processed of vegetable protein composition, more particularly to a kind of preparation side of samara oligopeptide
Method.
Background technology
Elaeagnus mollis benevolence is rich in grease, protein, multivitamin and mineral matter, and the wherein content of protein can reach
To 32.21%, contained protein is made up of albumin, globulin, alcohol soluble protein, glutelin in elaeagnus mollis benevolence, with paddy
The content highest of albumen, about 65.92-68.27% contain 17 kinds of amino acid in albumen, 8 kinds necessary to human body and animal
Amino acid contains 7 kinds.Substantial amounts of accessory substance samara crude protein is had during samara oil extraction to be formed, and can be that samara is oligomeric
The production of peptide provides raw material sources, and new outlet is provided for the production that becomes more meticulous of samara.
Amino acid forms peptide by peptide key connection, and it is the ultimate constituent of body tissue cell.Simplest peptide is
It is made up of two amino acid；The straight key peptide formed by 2~10 amino acid by peptide bond is referred to as oligopeptides or small peptide；More than 10
The condensate that individual amino acid is got up by peptide linkage is then referred to as polypeptide.Scientist is found that 100 that planting biology lives more in human body
Property peptide, these peptides have the function that transmit physiologic information, regulation of physiological functions, for the nerve of human body, digestion, reproduction, growth,
The maintenance of the system normal physiological activities such as motion, metabolism, circulation is extremely important, generation, development, treatment, the rehabilitation of all diseases
All there is relation with polypeptide.
Samara oligopeptide is using samara albumen as substrate, so that the molecular weight obtained after corresponding protease hydrolytic is small and has
The peptide molecule of certain activity.Rich in glutamic acid, proline, aspartic acid, arginine etc. in samara oligopeptide.Special amino
Acid composition and connected mode assign samara oligopeptide multiple biological activities, such as：It is anti-oxidant, immunological regulation, protect liver, antitumor, anti-
Inflammation, antibacterial, function antifatigue, that particularly there is alleviation Menopause symptom.
The content of the invention
It is an object of the invention to provide a kind of preparation method of samara oligopeptide, is produced during being extracted to samara oil
Raw a large amount of accessory substances have carried out effective utilization, provide raw material sources for the production of samara oligopeptide, while be also the essence of samara
Refinement production provides new outlet.
To achieve the above object, the technical solution used in the present invention is：A kind of preparation method of samara oligopeptide is provided,
Specifically comprise the following steps：
（1）Samara crude protein is dissolved in distilled water, solid-liquid ratio is（1~5）︰ 9, with homogenizer homogeneous, homogenizing time 10
Min~50min；It is 8.5~9.0 with NaOH regulations pH, temperature is 45 DEG C~55 DEG C, single by every gram of protein 2 000~5000
Position enzyme dosage addition alkali protease, digests 2~2.5h, enzyme deactivation, the time is 10 min~15min；
（2）Step（1）After end, in being centrifuged on centrifuge, rotating speed is 3000~6000 turns/min, the min of centrifugation time 10~
20min, take supernatant；
（3）Secondary enzymolysis, step（2）After the completion of, it is 60%~90% ethanol solution that the solid formation after centrifugation, which is dissolved in concentration, solid-liquid ratio
1~5:9.10 min of homogenizer homogeneous~50min, HCL regulation pH to 7.0, add aspergillus oryzae protease, papain, in
Property one or both of protease, temperature control digests 0.5~3h, 10 min of enzyme deactivation~15min at 30 DEG C~50 DEG C.In
Centrifuge, rotating speed are 3000~6000r/min, 10 min of centrifugation time~20min, take supernatant；
（4）Step（3）After the completion of, merge supernatant, from 500 u~1000u milipore filter ultrafiltration, obtain molecular weight less than 1
000 u filtered solution, that is, obtain the rough liquid of samara oligopeptide, and spray-dried or freeze-drying obtains samara oligopeptide dry powder.
The oligomeric Gly-His-Lys molecular weight of samara prepared by the present invention is concentrated within 1000Da.
The beneficial effects of the invention are as follows：Samara albumen is digested from two kinds of solvents of pure water and ethanol solution, energy
The enough more efficient and thoroughly peptide by samara proteolysis into small molecule, mean molecule quantity 1000Da.
Embodiment
Embodiment 1
（1）Samara crude protein is dissolved in distilled water, solid-liquid ratio 1:5, with homogenizer homogeneous, homogenizing time is 10 min；
It is 9.0 with NaOH regulations pH, temperature is 45 DEG C, adds alkali protease by every gram of unit enzyme dosage of protein 5000, digests 2h,
Enzyme deactivation, time 15min；
（2）Step（1）After end, in being centrifuged on centrifuge, rotating speed is 3000 turns/min, centrifugation time 20min, takes supernatant；
（3）Secondary enzymolysis, step（2）After the completion of, it is 90% ethanol solution that the solid formation after centrifugation, which is dissolved in concentration, solid-liquid ratio 5:9.
Homogenizer homogeneous 10 min, HCL regulation pH to 7.0, adds neutral proteinase, temperature control digests 0.5h, enzyme deactivation 10 at 30 DEG C
min.In centrifuge, rotating speed is 3000 turns/min, centrifugation time 10min, takes supernatant；
（4）Step（3）After the completion of, merge supernatant, from 500-1000u milipore filter ultrafiltration, obtain molecular weight less than 1 000
U filtered solution, that is, obtain the rough liquid of samara oligopeptide, and spray-dried or freeze-drying obtains samara oligopeptide dry powder.
The oligomeric Gly-His-Lys molecular weight of samara prepared by the present invention is concentrated within 1000 Da.
Embodiment 2
（1）Samara crude protein is dissolved in distilled water, solid-liquid ratio 1:9, with homogenizer homogeneous, homogenizing time 50min；With
NaOH regulations pH is 8.5, and temperature is 55 DEG C, and alkali protease, enzymolysis are added by every gram of unit enzyme dosage of protein 2 000
2.5h, enzyme deactivation, time 10min；
（2）Step（1）After end, in being centrifuged on centrifuge, rotating speed is 6000 turns/min, centrifugation time 10min, takes supernatant；
（3）Secondary enzymolysis, step（2）After the completion of, it is 60% ethanol solution that the solid formation after centrifugation, which is dissolved in concentration, solid-liquid ratio 1:9.
Homogenizer homogeneous 50min, HCL regulation pH to 7.0, adds papain, temperature control digests 3h, enzyme deactivation 5min at 50 DEG C.
In centrifuge, rotating speed is 6000 turns/min, centrifugation time 20min, takes supernatant；
（4）Step（3）After the completion of, merge supernatant, from 500-1000u milipore filter ultrafiltration, obtain molecular weight less than 1 000
U filtered solution, that is, obtain the rough liquid of samara oligopeptide, and spray-dried or freeze-drying obtains samara oligopeptide dry powder.
The oligomeric Gly-His-Lys molecular weight of samara prepared by the present invention is concentrated within 1000 Da.
Embodiment 3
（1）Samara crude protein is dissolved in distilled water, solid-liquid ratio 3:9, with homogenizer homogeneous, homogenizing time 30min；With
NaOH regulations pH is 8.8, and temperature is 50 DEG C, adds alkali protease by every gram of unit enzyme dosage of protein 3000, digests 2.3h,
Enzyme deactivation, time 13min；
（2）Step（1）After end, in being centrifuged on centrifuge, rotating speed is 4500 turns/min, centrifugation time 15min, takes supernatant；
（3）Secondary enzymolysis, step（2）After the completion of, it is 75% ethanol solution that the solid formation after centrifugation, which is dissolved in concentration, solid-liquid ratio 2:9.
Homogenizer homogeneous 30min, HCL regulation pH to 7.0, adds complex enzyme（Papain：Neutral proteinase 1:2）Temperature control
At 40 DEG C, 2h, enzyme deactivation 12min are digested.In centrifuge, rotating speed is 4500 turns/min, centrifugation time 15min, takes supernatant；
（4）Step（3）After the completion of, merge supernatant, from 500-1000u milipore filter ultrafiltration, obtain molecular weight less than 1 000
U filtered solution, that is, obtain the rough liquid of samara oligopeptide, and spray-dried or freeze-drying obtains samara oligopeptide dry powder.
The oligomeric Gly-His-Lys molecular weight of samara prepared by the present invention is concentrated within 1000 Da.
Claims (2)
1. a kind of preparation method of samara oligopeptide, it is characterised in that specifically comprise the following steps：
（1）Samara crude protein is dissolved in distilled water, solid-liquid ratio is 1~5 ︰ 9, with homogenizer homogeneous, homogenizing time 10
Min~50min；It is 8.5~9.0 with NaOH regulations pH, temperature is 45 DEG C~55 DEG C, single by every gram of protein 2 000~5000
Position enzyme dosage addition alkali protease, digests 2~2.5h, enzyme deactivation, the time is 10 min~15min；
（2）Step（1）After end, in being centrifuged on centrifuge, rotating speed is 3000~6000 turns/min, the min of centrifugation time 10~
20min, take supernatant；
（3）Secondary enzymolysis, step（2）After the completion of, it is 60%~90% ethanol solution that the solid formation after centrifugation, which is dissolved in concentration, solid-liquid ratio
1~5:9；10 min of homogenizer homogeneous~50min, HCL regulation pH to 7.0, add aspergillus oryzae protease, papain, in
Property one or both of protease, temperature control digests 0.5~3h, 10 min of enzyme deactivation~15min at 30 DEG C~50 DEG C；In
Centrifuge, rotating speed are 3000~6000r/min, 10 min of centrifugation time~20min, take supernatant；
（4）Step（3）After the completion of, merge supernatant, from 500 u~1000u milipore filter ultrafiltration, obtain molecular weight less than 1
000 u filtered solution, that is, obtain the rough liquid of samara oligopeptide, and spray-dried or freeze-drying obtains samara oligopeptide dry powder.
A kind of 2. preparation method of samara oligopeptide according to claim 1, it is characterised in that：The drying, it can be selected
Spray drying and freeze-drying in any one.
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| CN201710901028.6A CN107460224A (en) | 2017-09-28 | 2017-09-28 | A kind of preparation method of samara oligopeptide |
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Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109762862A (en) * | 2019-03-04 | 2019-05-17 | 武汉百理王生物工程有限公司 | A kind of preparation method of high-purity lotus seed protein oligopeptide |
| CN111334550A (en) * | 2020-03-25 | 2020-06-26 | 李淑华 | Method for extracting small molecule peptide from winged nut |
| CN115517313A (en) * | 2021-06-24 | 2022-12-27 | 山西大学 | Method for improving functional properties of samara protein |
| CN117551174A (en) * | 2023-08-14 | 2024-02-13 | 北京康美禾源健康科技有限公司 | Polypeptide derived from samara seed kernel oil meal and auxiliary anti-inflammatory and antioxidant blood lipid reducing composition |
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| CA2508219C (en) * | 2002-12-24 | 2012-12-18 | Her Majesty The Queen In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada | Ace inhibitory peptides from plant materials |
| CN104365991A (en) * | 2014-11-17 | 2015-02-25 | 乳山市华隆生物科技有限公司 | Micromolecule peanut peptide powder with high protein content and preparation method thereof |
| CN105936927A (en) * | 2016-05-19 | 2016-09-14 | 杏辉天力（杭州）药业有限公司 | Walnut oligopeptide, and preparation technology and use thereof |
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2017
- 2017-09-28 CN CN201710901028.6A patent/CN107460224A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CA2508219C (en) * | 2002-12-24 | 2012-12-18 | Her Majesty The Queen In Right Of Canada As Represented By The Minister Of Agriculture And Agri-Food Canada | Ace inhibitory peptides from plant materials |
| CN104365991A (en) * | 2014-11-17 | 2015-02-25 | 乳山市华隆生物科技有限公司 | Micromolecule peanut peptide powder with high protein content and preparation method thereof |
| CN105936927A (en) * | 2016-05-19 | 2016-09-14 | 杏辉天力（杭州）药业有限公司 | Walnut oligopeptide, and preparation technology and use thereof |
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Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109762862A (en) * | 2019-03-04 | 2019-05-17 | 武汉百理王生物工程有限公司 | A kind of preparation method of high-purity lotus seed protein oligopeptide |
| CN109762862B (en) * | 2019-03-04 | 2023-01-24 | 武汉百理王生物工程有限公司 | Preparation method of high-purity lotus seed protein oligopeptide |
| CN111334550A (en) * | 2020-03-25 | 2020-06-26 | 李淑华 | Method for extracting small molecule peptide from winged nut |
| CN115517313A (en) * | 2021-06-24 | 2022-12-27 | 山西大学 | Method for improving functional properties of samara protein |
| CN115517313B (en) * | 2021-06-24 | 2024-03-12 | 山西大学 | Method for improving functional properties of samara protein |
| CN117551174A (en) * | 2023-08-14 | 2024-02-13 | 北京康美禾源健康科技有限公司 | Polypeptide derived from samara seed kernel oil meal and auxiliary anti-inflammatory and antioxidant blood lipid reducing composition |
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Application publication date: 20171212 |