CN110129394B - Peony peptide with anti-fatigue effect and preparation method thereof - Google Patents
Peony peptide with anti-fatigue effect and preparation method thereof Download PDFInfo
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- CN110129394B CN110129394B CN201910295625.8A CN201910295625A CN110129394B CN 110129394 B CN110129394 B CN 110129394B CN 201910295625 A CN201910295625 A CN 201910295625A CN 110129394 B CN110129394 B CN 110129394B
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- 235000006484 Paeonia officinalis Nutrition 0.000 title claims abstract description 174
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 84
- 230000002929 anti-fatigue Effects 0.000 title claims abstract description 45
- 238000002360 preparation method Methods 0.000 title claims abstract description 20
- 230000000694 effects Effects 0.000 title claims description 29
- 244000170916 Paeonia officinalis Species 0.000 title 1
- 241000736199 Paeonia Species 0.000 claims abstract description 174
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 123
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 123
- 239000004365 Protease Substances 0.000 claims abstract description 54
- KDXKERNSBIXSRK-UHFFFAOYSA-N Lysine Natural products NCCCCC(N)C(O)=O KDXKERNSBIXSRK-UHFFFAOYSA-N 0.000 claims abstract description 20
- 239000004472 Lysine Substances 0.000 claims abstract description 20
- KDXKERNSBIXSRK-YFKPBYRVSA-N L-lysine Chemical compound NCCCC[C@H](N)C(O)=O KDXKERNSBIXSRK-YFKPBYRVSA-N 0.000 claims abstract description 19
- FFEARJCKVFRZRR-BYPYZUCNSA-N L-methionine Chemical compound CSCC[C@H](N)C(O)=O FFEARJCKVFRZRR-BYPYZUCNSA-N 0.000 claims abstract description 19
- COLNVLDHVKWLRT-QMMMGPOBSA-N L-phenylalanine Chemical compound OC(=O)[C@@H](N)CC1=CC=CC=C1 COLNVLDHVKWLRT-QMMMGPOBSA-N 0.000 claims abstract description 19
- 229930182817 methionine Natural products 0.000 claims abstract description 19
- COLNVLDHVKWLRT-UHFFFAOYSA-N phenylalanine Natural products OC(=O)C(N)CC1=CC=CC=C1 COLNVLDHVKWLRT-UHFFFAOYSA-N 0.000 claims abstract description 19
- 108010004032 Bromelains Proteins 0.000 claims abstract description 18
- 108090000270 Ficain Proteins 0.000 claims abstract description 18
- 108010030544 Peptidyl-Lys metalloendopeptidase Proteins 0.000 claims abstract description 18
- 108090000787 Subtilisin Proteins 0.000 claims abstract description 18
- 150000001413 amino acids Chemical class 0.000 claims abstract description 18
- 235000019835 bromelain Nutrition 0.000 claims abstract description 18
- 235000019836 ficin Nutrition 0.000 claims abstract description 18
- POTUGHMKJGOKRI-UHFFFAOYSA-N ficin Chemical compound FI=CI=N POTUGHMKJGOKRI-UHFFFAOYSA-N 0.000 claims abstract description 18
- 235000012054 meals Nutrition 0.000 claims abstract description 18
- 108090000317 Chymotrypsin Proteins 0.000 claims abstract description 14
- 229960002376 chymotrypsin Drugs 0.000 claims abstract description 14
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 claims description 28
- 238000004108 freeze drying Methods 0.000 claims description 26
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 17
- 229920000858 Cyclodextrin Polymers 0.000 claims description 15
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 claims description 15
- 229910002092 carbon dioxide Inorganic materials 0.000 claims description 14
- 239000001569 carbon dioxide Substances 0.000 claims description 14
- 238000009835 boiling Methods 0.000 claims description 13
- 239000012153 distilled water Substances 0.000 claims description 13
- 238000010438 heat treatment Methods 0.000 claims description 13
- 239000006228 supernatant Substances 0.000 claims description 13
- 108091005804 Peptidases Proteins 0.000 claims description 12
- 235000019419 proteases Nutrition 0.000 claims description 12
- 102000004190 Enzymes Human genes 0.000 claims description 6
- 108090000790 Enzymes Proteins 0.000 claims description 6
- 108090000526 Papain Proteins 0.000 claims description 6
- 229940088598 enzyme Drugs 0.000 claims description 6
- 235000019834 papain Nutrition 0.000 claims description 6
- 229940055729 papain Drugs 0.000 claims description 6
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims 3
- 235000018102 proteins Nutrition 0.000 abstract description 114
- 235000001014 amino acid Nutrition 0.000 abstract description 17
- 238000000034 method Methods 0.000 abstract description 6
- 238000000354 decomposition reaction Methods 0.000 abstract description 4
- WHUUTDBJXJRKMK-UHFFFAOYSA-N Glutamic acid Natural products OC(=O)C(N)CCC(O)=O WHUUTDBJXJRKMK-UHFFFAOYSA-N 0.000 abstract description 3
- 235000013922 glutamic acid Nutrition 0.000 abstract description 3
- 239000004220 glutamic acid Substances 0.000 abstract description 3
- 102000007079 Peptide Fragments Human genes 0.000 abstract description 2
- 108010033276 Peptide Fragments Proteins 0.000 abstract description 2
- 239000000419 plant extract Substances 0.000 abstract description 2
- 239000002131 composite material Substances 0.000 abstract 1
- 125000000291 glutamic acid group Chemical group N[C@@H](CCC(O)=O)C(=O)* 0.000 abstract 1
- 230000000052 comparative effect Effects 0.000 description 17
- 238000001694 spray drying Methods 0.000 description 13
- 102000035195 Peptidases Human genes 0.000 description 9
- 230000009182 swimming Effects 0.000 description 5
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 230000007071 enzymatic hydrolysis Effects 0.000 description 4
- 238000006047 enzymatic hydrolysis reaction Methods 0.000 description 4
- 239000004475 Arginine Substances 0.000 description 3
- ODKSFYDXXFIFQN-UHFFFAOYSA-N arginine Natural products OC(=O)C(N)CCCNC(N)=N ODKSFYDXXFIFQN-UHFFFAOYSA-N 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 235000019633 pungent taste Nutrition 0.000 description 3
- ODKSFYDXXFIFQN-BYPYZUCNSA-P L-argininium(2+) Chemical compound NC(=[NH2+])NCCC[C@H]([NH3+])C(O)=O ODKSFYDXXFIFQN-BYPYZUCNSA-P 0.000 description 2
- 241000699670 Mus sp. Species 0.000 description 2
- 125000000539 amino acid group Chemical group 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 239000012634 fragment Substances 0.000 description 2
- 238000001727 in vivo Methods 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 230000035764 nutrition Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 235000015112 vegetable and seed oil Nutrition 0.000 description 2
- 102000029816 Collagenase Human genes 0.000 description 1
- 108060005980 Collagenase Proteins 0.000 description 1
- 241000196324 Embryophyta Species 0.000 description 1
- 102000009127 Glutaminase Human genes 0.000 description 1
- 108010073324 Glutaminase Proteins 0.000 description 1
- DGAQECJNVWCQMB-PUAWFVPOSA-M Ilexoside XXIX Chemical compound C[C@@H]1CC[C@@]2(CC[C@@]3(C(=CC[C@H]4[C@]3(CC[C@@H]5[C@@]4(CC[C@@H](C5(C)C)OS(=O)(=O)[O-])C)C)[C@@H]2[C@]1(C)O)C)C(=O)O[C@H]6[C@@H]([C@H]([C@@H]([C@H](O6)CO)O)O)O.[Na+] DGAQECJNVWCQMB-PUAWFVPOSA-M 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 108010059712 Pronase Proteins 0.000 description 1
- 241000218201 Ranunculaceae Species 0.000 description 1
- 229910000831 Steel Inorganic materials 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 238000007792 addition Methods 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 239000003963 antioxidant agent Substances 0.000 description 1
- 230000003078 antioxidant effect Effects 0.000 description 1
- 239000006227 byproduct Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 229960002424 collagenase Drugs 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 229920001184 polypeptide Polymers 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 102000004196 processed proteins & peptides Human genes 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 150000003839 salts Chemical class 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 239000010959 steel Substances 0.000 description 1
- 238000012360 testing method Methods 0.000 description 1
Classifications
-
- 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
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/54—Improvements relating to the production of bulk chemicals using solvents, e.g. supercritical solvents or ionic liquids
Abstract
The invention relates to the field of plant extracts, and particularly relates to an anti-fatigue peony peptide and a preparation method thereof, wherein the peony peptide comprises 11-16 amino acids, and contains one or more of lysine, phenylalanine and methionine. The invention utilizes a two-step decomposition method, selects nonspecific ficin and subtilisin for composite treatment, and firstly breaks peptide bonds at main glutamic acid positions in peony seed meal protein to form protein peptide fragments. Then, a small amount of nonspecific bromelain, specific chymotrypsin and endopeptidase Lys-C are utilized to retain lysine, phenylalanine and methionine in peptide chain molecules as much as possible, so that the anti-fatigue capability of the protein peptide is improved.
Description
Technical Field
The invention relates to the field of plant extracts, and particularly relates to peony peptide with an anti-fatigue effect and a preparation method thereof.
Background
Peony is a plant of Ranunculaceae and Paeonia, and is called "Tianxiang" in national color. The peony seed oil developed from peony seeds is a natural pollution-free food integrating nutrition and health care. The protein content of the peony seed meal which is one of the byproducts of the squeezed peony seed oil reaches 19.03 percent.
The protein is an important component of food, the protein is subjected to enzymolysis to obtain protein peptide, the functional characteristics and biological characteristics of the protein peptide are greatly different from those of the original protein, for example, the functional characteristics of the protein such as solubility, emulsibility and foamability can be improved after enzymolysis, China has a large amount of oil refined peony seed meal products every year, and a large amount of peony protein can be obtained, so that how to obtain the peony protein peptide meeting the needs of life with high efficiency and low cost is a research hotspot at present.
The anti-fatigue amino acid is used as a necessary influence component of human nutrition, and when a human body feels fatigue, inorganic salts such as sodium, potassium and the like are required to be supplemented, and necessary amino acid is also required to be supplemented. The peony peptide is obtained by adopting a peony seed protease enzymolysis method at present, and the obtained protein peptide has rich amino acid residues, has a good anti-fatigue effect, can remove free radicals in vivo after sports or labor and effectively relieves fatigue. The existing protein peptide has poor anti-fatigue effect and slow action.
Disclosure of Invention
The invention aims to overcome the problems in the prior art and provides a peony peptide with an anti-fatigue effect.
The invention also aims to provide a preparation method of the peony peptide with the anti-fatigue effect.
The purpose of the invention is realized by the following technical scheme:
a peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
A peony protein peptide with anti-fatigue effect is obtained by enzymolysis of peony protein, contains amino acid residues, has antioxidant capacity, can remove free radicals in vivo, and can relieve fatigue of human body. The peony peptide contains 11-16 amino acids, has a molecular weight of 1000Da-1500Da, has good water solubility, and improves the application range of the peony peptide. And a specific enzymolysis method is adopted, the peony peptide contains one or more of lysine, phenylalanine, methionine lysine, phenylalanine and methionine, and the amino acids have certain reducibility, so that the obtained peony peptide has a good anti-fatigue effect.
A preparation method of peony peptide with anti-fatigue effect is characterized by comprising the following steps: s1 extracting peony protein from peony seed meal; s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide; s3, adding distilled water into the peony protein obtained in the previous step according to the mass ratio of 1: 20-25 to dissolve the peony protein, adjusting the temperature to 37-40 ℃, and adjusting the pH value of the solution to 8-10; s4, adding protease accounting for 2-3% of the weight of the peony protein in the step, and carrying out enzymolysis for 30-50 min; s5, heating the solution subjected to enzymolysis to 100 ℃, keeping the temperature for 10-15 min, and then recovering to 50-55 ℃; s6, adding enzyme which accounts for 6-8% of the mass of the peony protein into the solution obtained in the step, carrying out enzymolysis for 2-2.5 h, and then boiling quickly; s7, centrifuging the solution obtained in the previous step, and freeze-drying the supernatant; s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out. S8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out. A preparation method of peony peptide with anti-fatigue effect adopts a two-step decomposition method. The peony seed meal protein contains nearly 25% of glutamic acid, and nonspecific ficin and subtilisin are firstly adopted to carry out enzymolysis on the peony seed protein. After enzymolysis, the peony seeds are subjected to enzymolysis to obtain a plurality of small fragments; then, a small amount of nonspecific bromelain is adopted to hydrolyze the glutaminase in the lower fragment obtained in the first step of enzymolysis as much as possible, specific papain and endopeptidase Lys-C are used for enzymolysis, and nonspecific bromelain is used for enzymolysis of peptide bonds between arginine and arginine as well as between arginine and lysine in peony seed protein. This allows the peptide chain to retain as much lysine, phenylalanine, methionine as possible. The enzyme amount and enzymolysis time added in the two-step decomposition are controlled, so that the anti-fatigue capability of the product can be improved more effectively.
The peony protein has certain pungency, the pungency of the peony protein can be removed by extracting the peony protein with supercritical carbon dioxide, and finally obtained peony representative polypeptide is embedded with cyclodextrin, so that the pungency of the peony protein can be removed more effectively.
Preferably, the protease in S4 is ficin and subtilisin with the ratio of 1: 2-2.5.
Preferably, the protease in S6 is bromelain, chymotrypsin and endopeptidase Lys-C with the ratio of 1-3: 2-5: 3-7.
Preferably, the ph of the solution is adjusted to 10 in step S3.
Preferably, the total amount of the protease added in the step S4 is 2.5% of the mass of the peony protein.
Preferably, the ratio of ficin to subtilisin in step S4 is 1:2, and the enzymolysis time is 40 min.
Preferably, the ratio of bromelain, papain and endopeptidase Lys-C in step S6 is 2:5: 7.
Preferably, the amount of protease added in step S6 is 7% of the mass of peony protein.
Preferably, the enzymolysis time in the step S6 is 1 h.
Compared with the prior art, the invention has the following technical effects:
a peony peptide with anti-fatigue effect and a preparation method thereof utilize a two-step enzymolysis method, firstly utilize nonspecific protease to carry out enzymolysis on peony seed protein to obtain small chain peptide, and then utilize characteristic protease to carry out further enzymolysis. The protein peptide obtained by enzymolysis has better anti-fatigue capability.
Detailed Description
In order to make the objects, technical solutions and advantages of the present invention more apparent, the technical solutions of the present invention will be described in detail below with reference to specific examples and comparative examples. It is to be understood that the described embodiments are merely exemplary of the invention, and not restrictive of the full scope of the invention. All other embodiments, which can be derived by a person skilled in the art from the examples given herein without any inventive step, are within the scope of the present invention.
Unless otherwise specified, the equipment used in this example, comparative example and experimental example was conventional experimental equipment, and the reagents used were commercially available.
Example 1
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:20 to dissolve the peony protein, adjusting the temperature to 37 ℃, and adjusting the pH value of the solution to 8;
s4 adding ficin and subtilisin with a ratio of 1:2, which is 2% of the mass of the peony protein, into the solution of the step S3, and carrying out enzymolysis for 30 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C which account for 6 percent of the weight of the peony protein and have the proportion of 1:2:3 into the step S5, performing enzymolysis for 2 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
Example 2
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:25 to dissolve the peony protein, adjusting the temperature to 40 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2.5 to 3% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 50 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 15min, and then recovering to 55 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C which account for 6 percent of the weight of the peony protein and have the ratio of 3:5:7 in the step S5, performing enzymolysis for 2.5 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
Example 3
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:5:7, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 2.2 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
Comparative example 1
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 60 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:5:7, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 2.2 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant; s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The enzymatic hydrolysis time in step S4 of this comparative example was longer than that of example 3.
Comparative example 2
A peony peptide with an anti-fatigue effect comprises 11-15 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 20 min; s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:5:7, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 2.2 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The enzymatic hydrolysis time in step S4 of this comparative example was shorter than that of example 3.
Comparative example 3
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal; s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:5:7, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 0.5h, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant; s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The enzymatic hydrolysis time in step S6 of this comparative example was shorter than that of example 3.
Comparative example 4
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:5:7, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 3 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant; s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The enzymatic hydrolysis time in step S6 of this comparative example was longer than that of example 3.
Comparative example 5
A peony peptide with an anti-fatigue effect comprises 6-9 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:3, 2.5% of the weight of the peony protein, into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, papain and endopeptidase Lys-C with the proportion of 2:5:7, wherein the weight of the peony protein is 7 percent in the step S5, carrying out enzymolysis for 2.2h, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
Compared with example 3, the proportion of subtilisin added in step S4 of this comparative example is higher, and chymotrypsin is replaced by papain in step S6.
Comparative example 6
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and pronase with a ratio of 1:2, which is 2.5% of the weight of the peony protein, into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:5:7, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 2.2 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The enzyme added in step S4 of this example was different compared to example 3.
Comparative example 7
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6, adding bromelain, chymotrypsin and endopeptidase Lys-C with the proportion of 2:7:3, wherein the mass of the peony protein is 7% in the step S5, performing enzymolysis for 2.2 hours, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The comparative example varied in the ratio of enzyme added in step S6 compared to example 3.
Comparative example 8
A peony peptide with an anti-fatigue effect comprises 11-16 amino acids, wherein the peony peptide contains one or more of lysine, phenylalanine and methionine.
The preparation method of the anti-fatigue peony peptide comprises the following steps:
s1 extracting peony protein from peony seed meal;
s2, extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3, adding distilled water into the peony protein obtained in the step S2 according to the mass ratio of 1:22 to dissolve the peony protein, adjusting the temperature to 39 ℃, and adjusting the pH value of the solution to 10;
s4 adding ficin and subtilisin with a ratio of 1:2 of 2.5% of the weight of the peony protein into the solution of the step S3, and carrying out enzymolysis for 40 min;
s5, heating the solution after enzymolysis in the step S4 to 100 ℃, keeping for 10min, and then recovering to 50 ℃;
s6 adding 7% of collagenase, papain and endopeptidase Lys-C with the proportion of 2:5:7 based on the weight of the peony protein in the step S5, carrying out enzymolysis for 2.2h, and then boiling quickly;
s7, centrifuging the solution obtained in the step S6, and freeze-drying the supernatant;
s8 the protein peptide obtained by S7 freeze-drying is dissolved again, cyclodextrin is added, and then spray-drying is carried out.
The comparative example, step S6, was different in protease than example 3.
Examples of the experiments
120 groups of mice with uniform weight are selected, and are randomly divided into 12 groups, and the feed of each group of mice is added with each example, each proportion and a blank control. The following experiments were performed after 2 months of feeding:
and (3) carrying out a load swimming experiment: loading a steel wire according to the weight of 5% of the weight of the mouse, putting the mouse into a water tank with the water temperature of 25 +/-1 ℃ for swimming, starting timing until the mouse sinks to the water bottom for 8 seconds and does not float any more, and recording the swimming time for finishing exhaustive swimming.
TABLE 1 mouse swimming time situation table
From the test results, the invention utilizes a two-step decomposition method, selects nonspecific ficin and subtilisin to compound, firstly breaks the peptide bond at the main glutamic acid in the peony seed meal protein, and forms the protein peptide fragment. Then, a small amount of nonspecific bromelain, specific chymotrypsin and endopeptidase Lys-C are utilized to retain lysine, phenylalanine and methionine in peptide chain molecules as much as possible, so that the anti-fatigue capability of the protein peptide is improved.
Those of skill in the art will recognize that such devices, apparatuses, devices, systems may be connected, arranged, or configured in any manner. Words such as "including," "comprising," "having," and the like are open-ended words that mean "including, but not limited to," and are used interchangeably therewith. The words "or" and "as used herein mean, and are used interchangeably with, the word" and/or, "unless the context clearly dictates otherwise. The word "such as" is used herein to mean, and is used interchangeably with, the phrase "such as but not limited to".
Also, as used herein, the use of "or" in a list of items beginning with "at least one" indicates a separate list, e.g., "A, B or at least one of C" means A or B or C, or AB or AC or BC, or ABC (i.e., A and B and C). Furthermore, the word "exemplary" does not mean that the described example is preferred or better than other examples.
The previous description of the disclosed aspects is provided to enable any person skilled in the art to make or use the present disclosure. Various modifications to these aspects will be readily apparent to those skilled in the art, and the generic principles defined herein may be applied to other aspects without departing from the scope of the disclosure. Thus, the present disclosure is not intended to be limited to the aspects shown herein but is to be accorded the widest scope consistent with the principles and novel features disclosed herein.
The foregoing description has been presented for purposes of illustration and description. Furthermore, this description is not intended to limit embodiments of the disclosure to the form disclosed herein. While a number of example aspects and embodiments have been discussed above, those of skill in the art will recognize certain variations, modifications, alterations, additions and sub-combinations thereof.
Claims (4)
1. The peony peptide with the anti-fatigue effect is characterized by comprising 11-16 amino acids, wherein the molecular weight is 1000Da-1500Da, and the peony peptide contains one or more of lysine, phenylalanine and methionine; the peony peptide is prepared by enzymolysis of peony protein, and the preparation method comprises the following steps:
s1: extracting peony protein from peony seed meal;
s2: extracting the peony protein obtained in the step S1 by using supercritical carbon dioxide;
s3: adding distilled water into the peony protein obtained in the step S2 according to a mass ratio of 1 (20-25) to dissolve the peony protein, adjusting the temperature to 37-40 ℃, and adjusting the pH value of the solution to 10;
s4: adding protease accounting for 2-3% of the weight of the peony protein in the step S3, and carrying out enzymolysis for 30-50 min; the protease is ficin and subtilisin with the proportion of 1 (2-2.5);
s5: heating the solution subjected to enzymolysis in the step S4 to 100 ℃, keeping the temperature for 10-15 min, and then recovering to 50-55 ℃;
s6: adding bromelain, chymotrypsin and endopeptidase-Lys-C with the proportion of (1-3) - (2-5) - (3-7) as enzyme accounting for 7% -8% of the mass of the peony protein into the solution in the step S5, wherein the enzymolysis time is 2.2-2.5 h, and then boiling quickly;
s7: centrifuging the solution of step S6, and lyophilizing the supernatant;
s8: the protein peptide obtained by S7 freeze-drying is dissolved again, added with cyclodextrin and then spray-dried.
2. The peony peptide with anti-fatigue effect according to claim 1, wherein the total amount of protease added in step S4 is 2.5% of the peony protein by mass.
3. The peony peptide with anti-fatigue effect according to claim 2, wherein the ratio of ficin to subtilisin in step S4 is 1:2, and the enzymolysis time is 40 min.
4. The peony peptide with anti-fatigue effect according to any one of claims 1, 2 or 3, wherein the enzymes in step S6 are bromelain, papain and endopeptidase Lys-C in a ratio of 2:5: 7.
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| CN103937865B (en) * | 2014-04-28 | 2015-04-08 | 李�杰 | Peony polypeptide as well as preparation method and application thereof |
| CN108740279A (en) * | 2018-06-25 | 2018-11-06 | 菏泽盛世天下网络科技有限公司 | A kind of preparation method of peony seeds polypeptide |
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| Publication number | Priority date | Publication date | Assignee | Title |
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| CN103937865B (en) * | 2014-04-28 | 2015-04-08 | 李�杰 | Peony polypeptide as well as preparation method and application thereof |
| CN108740279A (en) * | 2018-06-25 | 2018-11-06 | 菏泽盛世天下网络科技有限公司 | A kind of preparation method of peony seeds polypeptide |
Non-Patent Citations (1)
| Title |
|---|
| Structure-activity Study of Macropin, a Novel Antimicrobial Peptide From the Venom of Solitary Bee Macropis Fulvipes (Hymenoptera: Melittidae);Lenka Monincová et al.;《J Pept Sci》;20140311;第20卷(第6期);摘要 * |
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