CN105624248A - Preparing method for alfalfa biological active peptide - Google Patents
Preparing method for alfalfa biological active peptide Download PDFInfo
- Publication number
- CN105624248A CN105624248A CN201610129481.5A CN201610129481A CN105624248A CN 105624248 A CN105624248 A CN 105624248A CN 201610129481 A CN201610129481 A CN 201610129481A CN 105624248 A CN105624248 A CN 105624248A
- Authority
- CN
- China
- Prior art keywords
- alfalfa
- enzymolysis
- biologically active
- active peptides
- enzyme
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 title claims abstract description 76
- 241000219823 Medicago Species 0.000 title claims abstract description 75
- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 56
- 238000000034 method Methods 0.000 title claims abstract description 31
- 102000004190 Enzymes Human genes 0.000 claims abstract description 37
- 108090000790 Enzymes Proteins 0.000 claims abstract description 37
- 239000000203 mixture Substances 0.000 claims abstract description 14
- 230000007062 hydrolysis Effects 0.000 claims abstract description 12
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 12
- 239000002994 raw material Substances 0.000 claims abstract description 11
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000001035 drying Methods 0.000 claims abstract description 9
- 239000012535 impurity Substances 0.000 claims abstract description 7
- 239000012141 concentrate Substances 0.000 claims abstract description 5
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 46
- 239000007788 liquid Substances 0.000 claims description 26
- HEMHJVSKTPXQMS-UHFFFAOYSA-M Sodium hydroxide Chemical compound [OH-].[Na+] HEMHJVSKTPXQMS-UHFFFAOYSA-M 0.000 claims description 18
- 238000002360 preparation method Methods 0.000 claims description 18
- 239000006071 cream Substances 0.000 claims description 16
- 239000000706 filtrate Substances 0.000 claims description 14
- MVORZMQFXBLMHM-QWRGUYRKSA-N Gly-His-Lys Chemical compound NCCCC[C@@H](C(O)=O)NC(=O)[C@@H](NC(=O)CN)CC1=CN=CN1 MVORZMQFXBLMHM-QWRGUYRKSA-N 0.000 claims description 13
- 108010038983 glycyl-histidyl-lysine Proteins 0.000 claims description 13
- VEXZGXHMUGYJMC-UHFFFAOYSA-N Hydrochloric acid Chemical compound Cl VEXZGXHMUGYJMC-UHFFFAOYSA-N 0.000 claims description 12
- 230000004071 biological effect Effects 0.000 claims description 12
- 239000006228 supernatant Substances 0.000 claims description 10
- 244000025254 Cannabis sativa Species 0.000 claims description 9
- 108090000623 proteins and genes Proteins 0.000 claims description 8
- 102000004169 proteins and genes Human genes 0.000 claims description 8
- 239000000758 substrate Substances 0.000 claims description 8
- 230000008719 thickening Effects 0.000 claims description 8
- 238000012216 screening Methods 0.000 claims description 7
- 238000001816 cooling Methods 0.000 claims description 6
- 230000008030 elimination Effects 0.000 claims description 6
- 238000003379 elimination reaction Methods 0.000 claims description 6
- 230000002255 enzymatic effect Effects 0.000 claims description 6
- 239000000463 material Substances 0.000 claims description 6
- 238000010298 pulverizing process Methods 0.000 claims description 6
- 230000001105 regulatory effect Effects 0.000 claims description 6
- 238000003756 stirring Methods 0.000 claims description 6
- 238000010792 warming Methods 0.000 claims description 6
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 abstract description 14
- 230000002000 scavenging effect Effects 0.000 abstract description 10
- 238000004519 manufacturing process Methods 0.000 abstract description 7
- 235000013305 food Nutrition 0.000 abstract description 2
- 102000004142 Trypsin Human genes 0.000 abstract 2
- 108090000631 Trypsin Proteins 0.000 abstract 2
- 230000009849 deactivation Effects 0.000 abstract 2
- 239000012588 trypsin Substances 0.000 abstract 2
- 108010033276 Peptide Fragments Proteins 0.000 abstract 1
- 102000007079 Peptide Fragments Human genes 0.000 abstract 1
- 239000002537 cosmetic Substances 0.000 abstract 1
- 230000008014 freezing Effects 0.000 abstract 1
- 238000007710 freezing Methods 0.000 abstract 1
- 239000000843 powder Substances 0.000 abstract 1
- 239000002002 slurry Substances 0.000 abstract 1
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 4
- 101710138460 Leaf protein Proteins 0.000 description 4
- 235000019750 Crude protein Nutrition 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000003078 antioxidant effect Effects 0.000 description 3
- 235000019658 bitter taste Nutrition 0.000 description 3
- 229960000935 dehydrated alcohol Drugs 0.000 description 3
- -1 flavonoid compound Chemical class 0.000 description 3
- 229920001184 polypeptide Polymers 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- OTEKOJQFKOIXMU-UHFFFAOYSA-N 1,4-bis(trichloromethyl)benzene Chemical compound ClC(Cl)(Cl)C1=CC=C(C(Cl)(Cl)Cl)C=C1 OTEKOJQFKOIXMU-UHFFFAOYSA-N 0.000 description 2
- 241000196324 Embryophyta Species 0.000 description 2
- 108010038807 Oligopeptides Proteins 0.000 description 2
- 102000015636 Oligopeptides Human genes 0.000 description 2
- 238000004332 deodorization Methods 0.000 description 2
- 238000005516 engineering process Methods 0.000 description 2
- 239000000796 flavoring agent Substances 0.000 description 2
- 235000019634 flavors Nutrition 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 239000012452 mother liquor Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 108010082495 Dietary Plant Proteins Proteins 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 241000219793 Trifolium Species 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 238000004440 column chromatography Methods 0.000 description 1
- 230000007812 deficiency Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000004090 dissolution Methods 0.000 description 1
- 238000005265 energy consumption Methods 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 229930003935 flavonoid Natural products 0.000 description 1
- 235000017173 flavonoids Nutrition 0.000 description 1
- 238000010438 heat treatment Methods 0.000 description 1
- 238000009776 industrial production Methods 0.000 description 1
- 235000008935 nutritious Nutrition 0.000 description 1
- 230000000050 nutritive effect Effects 0.000 description 1
- 235000019629 palatability Nutrition 0.000 description 1
- 239000000419 plant extract Substances 0.000 description 1
- 238000000751 protein extraction Methods 0.000 description 1
- 150000003254 radicals Chemical class 0.000 description 1
- 239000013558 reference substance Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 238000005507 spraying Methods 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
Classifications
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Health & Medical Sciences (AREA)
- Zoology (AREA)
- Engineering & Computer Science (AREA)
- Wood Science & Technology (AREA)
- Microbiology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Biotechnology (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Peptides Or Proteins (AREA)
Abstract
A preparing method for alfalfa biological active peptide comprises the steps that fresh or dry alfalfa herb serves as raw materials, and is screened to remove impurities, cleaned, drained, ground and added with water for slurry, and a raw material solution is obtained; the raw material solution is added with trypsin for enzymolysis; enzyme deactivation treatment is carried out after enzymolysis is finished, a mixture solution obtained after enzyme deactivation is centrifuged or filtered, and supernate or filter liquor is obtained; the supernate or filter liquor is subjected to vacuum concentration treatment, and alfalfa biological active peptide paste is obtained; the alfalfa biological active peptide paste is subjected to freezing drying treatment, and alfalfa biological active peptide powder is obtained. Compared with the prior art, according to the preparing method, alfalfa is directly subjected to enzymolysis with trypsin, the process is easy and convenient, the production cost is low, the peptide yield is high, the degree of hydrolysis can reach 32.56%, the peptide fragment component mainly concentrates at 1000 Da or lower, and the prepared alfalfa biological active peptide has high DPPH. scavenging activity, and can be applied and popularized in the fields such as food, health-care products, cosmetics and feed processing.
Description
Technical field
The invention belongs to the preparation field of plant origin biologically active peptides, the preparation method of especially a kind of alfalfa biologically active peptides.
Background technology
Alfalfa system pulse family, belong to per nnial herb, it is the herbage planting the widest, cultivation in the world the earliest, there is output height, good palatability, the feature such as nutritious so be described as " king of herbage ", generally plant in areas such as China northeast, North China, northwests. The amino acid kind of alfalfa is comparatively complete and balanced, and nutritive value is higher, is a kind of excellent vegetable protein resource.
At present, the aspect such as extraction research of alfalfa mainly being concentrated on to breeding, plantation, screening and flavonoid compound, leaf protein both at home and abroad, in prior art, research to alfalfa biologically active peptides is little. Wherein, the top condition that hydrolysis by novo alfalfa leaf protein prepares oligopeptides has been studied by Zhang Yong etc., and under top condition, degree of hydrolysis is 27.50%, and the average peptide chain length of oligopeptides is 3.64(China feed, 2007,18:15-17); The open specification sheets CN104195204A of Chinese invention patent application discloses the preparation method of a kind of alfalfa polypeptide, heating method is adopted to obtain alfalfa crude protein, again through alcohol steep decoloration deodorization, afterwards with Protamex enzyme and Flavourzyme1000L enzyme enzymolysis alfalfa crude protein, alfalfa polypeptide is obtained finally by spraying dry, content of peptides reaches 70%��75%, and the Gly-His-Lys of gained, without bitterness sense, can directly be taken. But above-mentioned two kinds of methods are all again its enzymolysis is carried out polypeptide preparation after first extracting leaf protein from clover grass, and leaf protein extraction yield is low, and energy consumption is big, and these two kinds of methods all exist the shortcoming that technique is numerous and diverse, production cost is high.
Summary of the invention
Technical problem to be solved by this invention is for the deficiencies in the prior art, it is provided that the preparation method of the alfalfa biologically active peptides that a kind of technique is easy, production cost is low, finished product has relatively strong anti-oxidative activity.
Technical problem to be solved by this invention is realized by following technical scheme. The present invention is the preparation method of a kind of alfalfa biologically active peptides, is characterized in, its step is as follows:
(1) Feedstock treating: get the fresh grass of alfalfa or hay, screening impurity elimination, clean, drain after drop into pulverizer and pulverize, add water homogenate than 1:9 ~ 1:11 by material quality after pulverizing, obtain stock liquid;
(2) enzymolysis: above-mentioned raw materials liquid pump is entered enzymatic vessel, pH is regulated with sodium hydroxide solution or hydrochloric acid soln, and constantly stir, in above-mentioned stock liquid, add trypsinase by enzyme concentration 800 ~ 1200U/g substrate protein and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 3 ~ 5.5h, hydrolysis temperature 45 ~ 60 DEG C, enzymolysis pH6 ~ 7.5;
(3) go out enzyme: enzymolysis is warming up to more than 90 DEG C after terminating, and keeps 10min, and go out enzyme, and cooling, carries out centrifugal treating to the mixture after the enzyme that goes out, obtain supernatant liquor, or is filtered by the mixture after the described enzyme that goes out, and obtains filtrate;
(4) vacuum concentration: above-mentioned clear liquid or filtrate suction concentration tank are carried out concentration, and thickening temperature is 50 ~ 65 DEG C, concentrates to final moisture content 40 ~ 60%, namely obtains alfalfa biologically active peptides cream.
(5) lyophilize: above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-60 ~-40 DEG C, is dried to final moisture content below 10%, namely obtains alfalfa biological activity Gly-His-Lys.
In the preparation method of alfalfa biologically active peptides of the present invention, preferred technical scheme or technology feature are further:
1, in step (1): during homogenate, the mass ratio of raw material and water is 1:10.
2, in step (2): add trypsinase by enzyme concentration 900U/g substrate protein in above-mentioned stock liquid and carry out enzyme, enzymolysis process parameter is: enzymolysis time 4h, hydrolysis temperature 55 DEG C, enzymolysis pH6.5.
3, in step (4): above-mentioned supernatant liquor or filtrate suction concentration tank are carried out concentration, and thickening temperature is 55 DEG C, concentrating to final moisture content is 50%, namely obtains alfalfa biologically active peptides cream.
4, in step (5): above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-50 DEG C, being dried to final moisture content is 8%, namely obtains alfalfa biological activity Gly-His-Lys.
Compared with prior art, alfalfa is directly carried out enzymolysis with trypsinase by the present invention, peptide yield height, degree of hydrolysis can reach 32.56%, peptide segment molecule amount mainly concentrates on below 1000Da, and obtained alfalfa biologically active peptides has stronger hexichol for bitter taste hydrazine base free radical (DPPH) Scavenging activity; Eliminating in preparation process and extract crude protein and decoloration deodorization step, not only simplify production technique, further reduces production cost, the use improving product is tired. The alfalfa biologically active peptides that the present invention obtains is natural plant extract, in light green, there is light green grass fragrant, local flavor is all good, there is stronger anti-oxidant activity simultaneously, can be applicable in fields such as food, healthcare products, makeup and feed manufacturing, and technique is easy, production cost is low, is applicable to large-scale industrial production.
Accompanying drawing explanation
Fig. 1 is that xitix (Vc) is to the measurement result of DPPH Scavenging activity;
Fig. 2 is that the alfalfa biological activity Gly-His-Lys adopting the method described in the embodiment of the present invention 4 to prepare is to the measurement result of DPPH Scavenging activity.
Embodiment
Referring to accompanying drawing, further describe the concrete technical scheme of the present invention, so that the technician of this area understands the present invention further, and do not form the restriction to its right.
Embodiment 1, the preparation method of a kind of alfalfa biologically active peptides, its step is as follows:
(1) Feedstock treating: get the fresh grass of alfalfa or hay, screening impurity elimination, clean, drain after drop into pulverizer and pulverize, add water homogenate than 1:9 by material quality after pulverizing, obtain stock liquid;
(2) enzymolysis: above-mentioned raw materials liquid pump is entered enzymatic vessel, pH is regulated with sodium hydroxide solution or hydrochloric acid soln, and constantly stir, in above-mentioned stock liquid, add trypsinase by enzyme concentration 800U/g substrate protein and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 3h, hydrolysis temperature 45 DEG C, enzymolysis pH6;
(3) go out enzyme: enzymolysis is warming up to more than 90 DEG C after terminating, and keeps 10min, and go out enzyme, and cooling, carries out centrifugal treating to the mixture after the enzyme that goes out, obtain supernatant liquor, or is filtered by the mixture after the described enzyme that goes out, and obtains filtrate;
(4) vacuum concentration: above-mentioned clear liquid or filtrate suction concentration tank are carried out concentration, and thickening temperature is 50 DEG C, concentrating to final moisture content is 40%, namely obtains alfalfa biologically active peptides cream.
(5) lyophilize: above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-60 DEG C, being dried to final moisture content is 10%, namely obtains alfalfa biological activity Gly-His-Lys.
Embodiment 2, the preparation method of a kind of alfalfa biologically active peptides, its step is as follows:
(1) Feedstock treating: get the fresh grass of alfalfa or hay, screening impurity elimination, clean, drain after drop into pulverizer and pulverize, add water homogenate than 1:11 by material quality after pulverizing, obtain stock liquid;
(2) enzymolysis: above-mentioned raw materials liquid pump is entered enzymatic vessel, pH is regulated with sodium hydroxide solution or hydrochloric acid soln, and constantly stir, in above-mentioned stock liquid, add trypsinase by enzyme concentration 1200U/g substrate protein and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 5.5h, hydrolysis temperature 60 DEG C, enzymolysis pH7.5;
(3) go out enzyme: enzymolysis is warming up to more than 90 DEG C after terminating, and keeps 10min, and go out enzyme, and cooling, carries out centrifugal treating to the mixture after the enzyme that goes out, obtain supernatant liquor, or is filtered by the mixture after the described enzyme that goes out, and obtains filtrate;
(4) vacuum concentration: above-mentioned clear liquid or filtrate suction concentration tank are carried out concentration, and thickening temperature is 65 DEG C, concentrating to final moisture content is 60%, namely obtains alfalfa biologically active peptides cream.
(5) lyophilize: above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-40 DEG C, being dried to final moisture content is 6%, namely obtains alfalfa biological activity Gly-His-Lys.
Embodiment 3, the preparation method of a kind of alfalfa biologically active peptides, its step is as follows:
(1) Feedstock treating: get the fresh grass of alfalfa or hay, screening impurity elimination, clean, drain after drop into pulverizer and pulverize, add water homogenate than 1:10 by material quality after pulverizing, obtain stock liquid;
(2) enzymolysis: above-mentioned raw materials liquid pump is entered enzymatic vessel, pH is regulated with sodium hydroxide solution or hydrochloric acid soln, and constantly stir, in above-mentioned stock liquid, add trypsinase by enzyme concentration 1000U/g substrate protein and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 4.5h, hydrolysis temperature 55 DEG C, enzymolysis pH7;
(3) go out enzyme: enzymolysis is warming up to more than 90 DEG C after terminating, and keeps 10min, and go out enzyme, and cooling, carries out centrifugal treating to the mixture after the enzyme that goes out, obtain supernatant liquor, or is filtered by the mixture after the described enzyme that goes out, and obtains filtrate;
(4) vacuum concentration: above-mentioned clear liquid or filtrate suction concentration tank are carried out concentration, and thickening temperature is 60 DEG C, concentrating to final moisture content is 50%, namely obtains alfalfa biologically active peptides cream.
(5) lyophilize: above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-50 DEG C, being dried to final moisture content is 6%, namely obtains alfalfa biological activity Gly-His-Lys.
Embodiment 4, the preparation method of a kind of alfalfa biologically active peptides, its step is as follows:
(1) Feedstock treating: get the fresh grass of alfalfa or hay, screening impurity elimination, clean, drain after drop into pulverizer and pulverize, add water homogenate than 1:10 by material quality after pulverizing, obtain stock liquid;
(2) enzymolysis: above-mentioned raw materials liquid pump is entered enzymatic vessel, pH is regulated with sodium hydroxide solution or hydrochloric acid soln, and constantly stir, in above-mentioned stock liquid, add trypsinase by enzyme concentration 900U/g substrate protein and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 4h, hydrolysis temperature 55 DEG C, enzymolysis pH6.5;
(3) go out enzyme: enzymolysis is warming up to more than 90 DEG C after terminating, and keeps 10min, and go out enzyme, and cooling, carries out centrifugal treating to the mixture after the enzyme that goes out, obtain supernatant liquor, or is filtered by the mixture after the described enzyme that goes out, and obtains filtrate;
(4) vacuum concentration: above-mentioned supernatant liquor or filtrate suction concentration tank are carried out concentration, and thickening temperature is 55 DEG C, concentrating to final moisture content is 50%, namely obtains alfalfa biologically active peptides cream.
(5) lyophilize: above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-50 DEG C, being dried to final moisture content is 8%, namely obtains alfalfa biological activity Gly-His-Lys.
The alfalfa biological activity Gly-His-Lys prepared according to the present embodiment method is light green, there is light green grass fragrant, local flavor is all good, has measured to its molecular weight and to the scavenging(action) of hexichol for bitter taste hydrazine base free radical (DPPH), and measuring method and result be:
1, the mensuration of molecular weight
Adopt dextrane gel (G-15) column chromatography to be detected by the molecular weight of alfalfa biologically active peptides, the results are shown in Table 1.
The molecular weight detection result of table 1. alfalfa biologically active peptides.
By result it will be seen that the molecular weight of alfalfa biologically active peptides mainly concentrates on 443��509Da, content accounts for 73.6%, and the small-molecular peptides of acquisition is more easily absorbed by body.
2, to the mensuration of DPPH scavenging(action).
DPPH method is adopted to evaluate the anti-oxidant activity of alfalfa biologically active peptides. After alfalfa biological activity Gly-His-Lys water dissolution, get that supernatant liquid is concentrated obtains the dense biologically active peptides concentrated solution of alfalfa, stand-by. Taking xitix (Vc) as positive reference substance, Vc and alfalfa biologically active peptides concentrated solution are mixed with dehydrated alcohol respectively the solution to be measured of different concns; It is 1 �� 10 that DPPH dehydrated alcohol is mixed with concentration-3The mother liquor of mol/L, deposits in 0��4 DEG C. During experiment, mother liquor is diluted to 1 �� 10-4After mol/L, get 2mL and join in 10mL colorimetric cylinder, then add the solution to be measured prepared, fully mix, at room temperature static 30min. At maximum wavelength 517nm place, making blank with dehydrated alcohol, measure each absorbancy (A), the parallel survey of each absorbancy 3 times, gets its mean value.
In formula: Ai: the absorbance adding the DPPH solution after solution to be measured;
Aj: the solvent of DPPH solution and the mixed absorbance of solution to be measured;
A0: the absorbance not adding the DPPH solution of solution to be measured.
Result shows, in certain mass concentration range, the Scavenging activity of DPPH is increased by alfalfa biologically active peptides with the increase of its mass concentration, and its Scavenging activity and mass concentration are good logarithm regression relation. The effective concentration removing reagent needed for 50%DPPH is defined as EC50, EC50It is worth more little, represents that the ability of scavenging free radicals is more strong. Through calculating it will be seen that the EC of xitix50Value is 0.033mg/mL, the EC of alfalfa biologically active peptides50Value is 2.22mg/mL, illustrates that alfalfa biological activity Gly-His-Lys has stronger DPPH Scavenging activity, and result is with reference to Fig. 1, Fig. 2.
Claims (5)
1. the preparation method of an alfalfa biologically active peptides, it is characterised in that, its step is as follows:
(1) Feedstock treating: get the fresh grass of alfalfa or hay, screening impurity elimination, clean, drain after drop into pulverizer and pulverize, add water homogenate than 1:9 ~ 1:11 by material quality after pulverizing, obtain stock liquid;
(2) enzymolysis: above-mentioned raw materials liquid pump is entered enzymatic vessel, pH is regulated with sodium hydroxide solution or hydrochloric acid soln, and constantly stir, in above-mentioned stock liquid, add trypsinase by enzyme concentration 800 ~ 1200U/g substrate protein and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 3 ~ 5.5h, hydrolysis temperature 45 ~ 60 DEG C, enzymolysis pH6 ~ 7.5;
(3) go out enzyme: enzymolysis is warming up to more than 90 DEG C after terminating, and keeps 10min, and go out enzyme, and cooling, carries out centrifugal treating to the mixture after the enzyme that goes out, obtain supernatant liquor, or is filtered by the mixture after the described enzyme that goes out, and obtains filtrate;
(4) vacuum concentration: above-mentioned clear liquid or filtrate suction concentration tank are carried out concentration, and thickening temperature is 50 ~ 65 DEG C, concentrates to final moisture content 40 ~ 60%, namely obtains alfalfa biologically active peptides cream;
(5) lyophilize: above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, and drying temperature is-60 ~-40 DEG C, is dried to final moisture content below 10%, namely obtains alfalfa biological activity Gly-His-Lys.
2. the preparation method of alfalfa biologically active peptides according to claim 1, it is characterised in that, in step (1): during homogenate, the mass ratio of raw material and water is 1:10.
3. the preparation method of alfalfa biologically active peptides according to claim 1, it is characterized in that, in step (2): add trypsinase by enzyme concentration 900U/g substrate protein in above-mentioned stock liquid and carry out enzymolysis, enzymolysis process parameter is: enzymolysis time 4h, hydrolysis temperature 55 DEG C, enzymolysis pH6.5.
4. the preparation method of alfalfa biologically active peptides according to claim 1, it is characterized in that, in step (4): above-mentioned supernatant liquor or filtrate suction concentration tank are carried out concentration, thickening temperature is 55 DEG C, concentrating to final moisture content is 50%, namely obtains alfalfa biologically active peptides cream.
5. the preparation method of alfalfa biologically active peptides according to claim 1, it is characterized in that, in step (5): above-mentioned alfalfa biologically active peptides cream is carried out lyophilize process, drying temperature is-50 DEG C, being dried to final moisture content is 8%, namely obtains alfalfa biological activity Gly-His-Lys.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610129481.5A CN105624248B (en) | 2016-03-08 | 2016-03-08 | Preparation method of alfalfa bioactive peptide |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610129481.5A CN105624248B (en) | 2016-03-08 | 2016-03-08 | Preparation method of alfalfa bioactive peptide |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN105624248A true CN105624248A (en) | 2016-06-01 |
| CN105624248B CN105624248B (en) | 2020-02-11 |
Family
ID=56039590
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610129481.5A Expired - Fee Related CN105624248B (en) | 2016-03-08 | 2016-03-08 | Preparation method of alfalfa bioactive peptide |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105624248B (en) |
Cited By (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106820113A (en) * | 2016-12-28 | 2017-06-13 | 陕西天宝大豆食品技术研究所 | Full clover peptide nutrient food and preparation method thereof |
| CN107319543A (en) * | 2017-07-18 | 2017-11-07 | 诸城市和生食品有限公司 | A kind of alfalfa cream and preparation method thereof |
| CN107446980A (en) * | 2017-09-26 | 2017-12-08 | 成都中医药大学 | A kind of Leaves of Hippophae L polypeptide and preparation method thereof |
| CN112807416A (en) * | 2021-01-18 | 2021-05-18 | 宁波御坊堂生物科技有限公司 | Traditional Chinese medicine bone peptide composition with function of improving bone joint health and preparation method thereof |
| KR102605467B1 (en) * | 2023-05-04 | 2023-11-23 | 한국콜마주식회사 | Cosmetic composition for skin improvement comprising an extract of alfalfa seed-derived glycoprotein |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104195204A (en) * | 2014-07-30 | 2014-12-10 | 内蒙古天奇生物科技有限公司 | Method for preparing alfalfa polypeptide |
-
2016
- 2016-03-08 CN CN201610129481.5A patent/CN105624248B/en not_active Expired - Fee Related
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104195204A (en) * | 2014-07-30 | 2014-12-10 | 内蒙古天奇生物科技有限公司 | Method for preparing alfalfa polypeptide |
Non-Patent Citations (2)
| Title |
|---|
| 宋执磊: "紫花苜蓿寡肽的制备及其利用性研究", 《中国学位论文全文数据库》 * |
| 谢正军等: "超滤对苜蓿蛋白酶解物抗氧化性的影响", 《江苏大学学报（自然科学版）》 * |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN106820113A (en) * | 2016-12-28 | 2017-06-13 | 陕西天宝大豆食品技术研究所 | Full clover peptide nutrient food and preparation method thereof |
| CN107319543A (en) * | 2017-07-18 | 2017-11-07 | 诸城市和生食品有限公司 | A kind of alfalfa cream and preparation method thereof |
| CN107446980A (en) * | 2017-09-26 | 2017-12-08 | 成都中医药大学 | A kind of Leaves of Hippophae L polypeptide and preparation method thereof |
| CN107446980B (en) * | 2017-09-26 | 2021-03-19 | 成都中医药大学 | Sea buckthorn leaf polypeptide and preparation method thereof |
| CN112807416A (en) * | 2021-01-18 | 2021-05-18 | 宁波御坊堂生物科技有限公司 | Traditional Chinese medicine bone peptide composition with function of improving bone joint health and preparation method thereof |
| KR102605467B1 (en) * | 2023-05-04 | 2023-11-23 | 한국콜마주식회사 | Cosmetic composition for skin improvement comprising an extract of alfalfa seed-derived glycoprotein |
Also Published As
| Publication number | Publication date |
|---|---|
| CN105624248B (en) | 2020-02-11 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105624248A (en) | Preparing method for alfalfa biological active peptide | |
| CN102078012B (en) | Application of Bacillus subtilis solid aminopeptidase in compound enzymolysis of porphyra | |
| CN104087639A (en) | Method for extracting micromolecular active substances from sunflower calathide | |
| CN107361241B (en) | Aquatic animal liver protective agent, preparation method thereof, feed and health-care product | |
| CN105037377A (en) | Extraction preparation method for sodium ferrous chlorophyllin and sodium ferrous chlorophyllin prepared through extraction preparation method | |
| CN103387599A (en) | Technology for extracting protein of folium mori | |
| CN106962590A (en) | A kind of method for preparing edible fungus protein powder | |
| CN105029632A (en) | Suaeda salsa fermented beverage and preparation method thereof | |
| CN111334540B (en) | Method for extracting dendrobium officinale polysaccharide by utilizing biological fermentation | |
| CN104418944A (en) | Technology for separating multiple bioactive components in maize germ | |
| CN103864944B (en) | A kind of preparation method of pholiota nameko polysaccharide crude product | |
| CN109380701A (en) | Utilize the method for soy sauce residues Production by Enzymes soy sauce matrix seasoning | |
| CN113797261A (en) | Preparation method of camellia flower/leaf extract | |
| CN104262500B (en) | It is a kind of that there is immunocompetent novel red support dictyophora fungus polysaccharide and its preparation method and application | |
| KR100827422B1 (en) | Manufacturing method for extract of chlorella | |
| CN108048432B (en) | Ginger protease extraction method | |
| KR101118156B1 (en) | Method for preparing em from stevia | |
| CN106086139A (en) | A kind of method utilizing fresh-water fishes noggin enzymolysis to prepare fish head polypeptides | |
| CN101503727B (en) | Hyriopsis cumingii enzymolysis polypeptide, as well as preparation and application thereof | |
| CN105266164A (en) | Manufacturing method of selenium-enriched functional water with high energy quantum | |
| CN108477350A (en) | A kind of peonidin tea and preparation method thereof | |
| CN105200107B (en) | The extracting method of Onchidium struma muscle crude protein | |
| CN103355727A (en) | Preparation method for solid Cordyceps militaris beverage | |
| CN103110059B (en) | Production method and product of crude kudzu root flour | |
| CN105037378A (en) | Method for preparing and extracting iron chlorophyll and prepared iron chlorophyll |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20180510 Address after: 222000 No. 8, Yu Chau Nan Road, Haizhou Development Zone, Haizhou District, Lianyungang, Jiangsu. Applicant after: Jiangsu Tianfulai Group Co.,Ltd. Address before: 222000 No. 8, Yu Chau Nan Road, Haizhou Development Zone, Haizhou District, Lianyungang, Jiangsu. Applicant before: Chen Yifeng |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20200211 |