CN108546281A - A kind of ginseng oligopeptide and its preparation method and application - Google Patents
A kind of ginseng oligopeptide and its preparation method and application Download PDFInfo
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- CN108546281A CN108546281A CN201810388562.6A CN201810388562A CN108546281A CN 108546281 A CN108546281 A CN 108546281A CN 201810388562 A CN201810388562 A CN 201810388562A CN 108546281 A CN108546281 A CN 108546281A
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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
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/17—Amino acids, peptides or proteins
- A23L33/18—Peptides; Protein hydrolysates
-
- 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
-
- 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
- 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
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
Abstract
The present invention is intended to provide a kind of ginseng oligopeptide and preparation method thereof, it is using ginseng as raw material, through biological enzymolysis, isolates and purifies final high productivity and obtain ginseng oligopeptide, this method is naturally environmentally friendly, any toxic chemical reagent is not added, the product yield of acquisition is high, and use easy to spread compensates for the blank in ginseng oligopeptide field.The present invention creatively proposes the preparation process of ginseng oligopeptide, compensates for the blank of domestic ginseng oligopeptide production, and the product yield that method using the present invention is prepared is high, the good, raciness of activity, has huge with foreground and market potential.
Description
Technical field：
The invention belongs to biological fields, are related to the production of oligopeptide, and in particular to a kind of ginseng oligopeptide and preparation method thereof.
Background technology：
Ginseng（Panax ginseng C. A. Mey）It is generally acknowledged both at home and abroad medicinal health treasure, belongs to Araliaceae
(Araliaceae) Panax PanaxL. is perennial herb plant, is known as " King of Herbs ".Ginseng is as a kind of
Rare traditional Chinese medicine possesses long medicinal history, for a long time except to ginsenoside etc., a few substance is studied extensively
Outside, certain research has also been carried out to ginseng protein, has been isolated and purified from ginseng and has provided the active polypeptide of important biomolecule
And protein.
Science in recent years thinks that absorption of human body protein is mainly to be absorbed in the form of peptide, and peptide is by amino acid group
At molecule, molecular weight between 5000-200Dalton, have extremely strong activity and diversity, can use peptide nutrition physiology work(
Can and bioactivity, adjust the physiological function of each system and cell in human body, activate related enzyme element in vivo, therefore, various peptides at
For the important substance of health.Oligopeptide is the catabolite of albumen, compared with albumen, polypeptide, has molecular weight smaller, water
Dissolubility is stronger, active higher, it is easier to the advantages such as human consumption's absorption.Usual oligopeptide has anti-oxidant, blood pressure lowering, inhibits blood
The physiological activity such as platelet aggregation activity, antitumor and immunological regulation, while also having and replenish the calcium, lose weight, beauty and skin care and delaying to decline
It waits for a long time functional characteristic.Currently, it is more to the extractions such as the composition of ginseng, saponin, saponin(e research both at home and abroad, but for ginseng oligopeptide
The research of peptide also gradually becomes more, such as Bao Lei etc. has inquired into the antifatigue effect of panax ginseng in jiljn oligopeptide（《Scientific and technological Leader》 2015
13 phases of year）, show that mouse movement endurance can be improved in the panax ginseng in jiljn oligopeptide intervention of suitable dosage, extend walking weight load,
The content of blood lactase acid and blood urea nitrogen after moving is reduced, the content of lactic acid dehydrogenase activity and hepatic glycogen is improved；He Lixia et al.
Have studied the immunoregulation effect of panax ginseng in jiljn oligopeptide（《Scientific and technological Leader》18 phases in 2015）, show panax ginseng in jiljn oligopeptide
Significantly improve ConA induction mice spleen lymphocytes proliferation ability, delayed allergy ability, antibody-producting cell number,
Mouse carbonic clearance index, the phagocytosis ratio and phagocytosis index of macrophage, NK cell activity（P<0.05）, and effect is better than whey egg
In vain.It follows that GOP can by enhance cellular immune function, humoral immune function, monocytes/macrophages phagocytic activity and
NK cell activity, plays the role of strengthen immunity；Bao Lei, Wang Junbo et al. have studied panax ginseng in jiljn oligopeptide to male mice
The influence of sexual function（《Chinese preventive medicine magazine》10 phases in 2015）, the results showed that each dosage group of panax ginseng in jiljn oligopeptide can
Different degrees of increase male mice accessory sex organ's index improves NO and testosterone concentration in serum, and it is latent to shorten mouse mating
Phase simultaneously increases number of copulations.
Although gradually increasing for the research of ginseng oligopeptide at present, effectively carried however, still lacking in the prior art
The method for taking ginseng oligopeptide, extremely rare foreign language datum is several to extract or synthesizes ginseng oligopeptide in chemical mode, but passes through
It investigates, the oligopeptide structure that above method obtains is single, the mainly six oligomeric peptide molecules of class oxidized form of glutathione class, and extracts
Rate is low, and the ginseng oligopeptide production cost of synthesis is high, and is eaten since the problems such as reagent remains is also not suitable for people.That is, existing
Still lack the effective method for obtaining ginseng oligopeptide in technology.
Invention content：
In view of the deficiencies of the prior art, the present invention is intended to provide a kind of ginseng oligopeptide and preparation method thereof, is original with ginseng
Material through biological enzymolysis, isolates and purifies final high productivity and obtains ginseng oligopeptide, and this method environmental protection naturally is not added any
The product yield of toxic chemical reagent, acquisition is high, and use easy to spread compensates for the blank in ginseng oligopeptide field.
In order to solve the above technical problems, the present invention uses following technological means：
A kind of preparation method of ginseng oligopeptide, which is characterized in that the preparation method of the ginseng oligopeptide includes the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.5-5.0 of ginseng slurries；
Step 2 is added cellulase in the ginseng slurries obtained to step 1, is digested 3-8 hours at a temperature of 40-45 DEG C,
PH is then adjusted to 8.0-9.0, temperature is adjusted to 25-30 DEG C, stirring and leaching 12-17 hours, then centrifuged with 3500rpm rotating speeds
20min takes supernatant；
The pH of supernatant made from step 2 is adjusted to 4-4.5 by step 3, and stirring, 12000rpm centrifuge 10min, discard
Clear liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, that is, be people
Join albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid is adjusted to 8.2-8.5, alkali protease is then added thereto and carries out first time enzyme
Solution, obtains first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 6.8-7.2, neutral protein is then added thereto by step 5
Enzyme and flavor protease carry out second and digest, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
Preferably, the preparation method of the ginseng oligopeptide includes the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.8 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 5 hours at a temperature of 42 DEG C, then
PH is adjusted to 8.5, temperature and is adjusted to 28 DEG C, stirring and leaching 15 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.2 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.2, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 7.0 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
Preferably, the preparation method of the ginseng oligopeptide includes the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.5 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 8 hours at a temperature of 40 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 25 DEG C, stirring and leaching 12 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant liquid,
Take precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng egg
In vain；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.2, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 6.8 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
It can be with, it is preferable that the preparation method of the ginseng oligopeptide includes the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 5.0 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 4 hours at a temperature of 45 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 30 DEG C, stirring and leaching 17 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.5 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.5, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 7.2 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
Preferably, the dosage of step 3 neutral and alkali protease is 550-700U/g, and the temperature of enzymolysis is 45-50 for the first time
DEG C, enzymolysis time 20-25min；The dosage of neutral proteinase is 200-300U/g in step 4, and the dosage of flavor protease is
The temperature of 700-900U/g, second of enzymolysis are 50-55 DEG C, enzymolysis time 60-120min.
Preferably, the filtering in step 6 includes coarse filtration and refined filtration, and coarse filtration uses aperture for the ceramic membrane of 400-1200nm
It carries out, refined filtration is carried out using the ceramic membrane of 50-200nm.
Based on above technical scheme, the invention has the advantages that and advantageous effect：
First, in view of the deficiencies of the prior art, the present invention proposes a kind of preparation methods of ginseng oligopeptide, and through party's legal system
It is standby to have obtained the oligomeric peptide product of ginseng of high-purity, high activity, the deficiency of ginseng oligopeptide preparation process is compensated for, through testing
Card is up to 87.5%, i.e., after the process of the present invention, greatly through ginseng oligopeptide of the present invention relative to the yield of ginseng protein
Partial ginseng protein is converted into ginseng oligopeptide, can greatly improve the utilization rate of ginseng protein, increases ginseng finishing
Economic value and added value.
Secondly, the present invention creatively uses cellulose broken wall and the heavy ginseng protein extraction work being combined of alkali soluble acid
Skill greatly improves the proposition efficiency of ginseng protein, sinks compared to only alkali soluble acid is carried out to ginseng slurries, using the above cellulose
Broken wall and the heavy ginseng protein extraction process being combined of alkali soluble acid, protein extraction amount improve 35.0%, especially alkali process and acid
The pH value of processing is that inventor obtains through a large number of experiments, and processing in the conditions of the invention can maximumlly obtain high-purity
Ginseng protein.
Again, the present invention creatively use alkali protease, neutral proteinase and flavor protease to ginseng protein into
Row step-by-step processing, and best treatment process parameter is determined, basic protein enzymatic treatment is first used, neutral proteinase is then used
With flavor protease processing, it can ensure that ginseng protein fully digests and generate oligopeptide, while can also efficiently reduce bitter peptides
Generation, the excellent flavor of final products, without the bitter taste of general oligomeric peptide product.The people being prepared using the method for the present invention
Ginseng oligopeptide is used directly for the production of food, health food or drug, the bitter taste without considering oligopeptide.
In conclusion the present invention creatively proposes the preparation process of ginseng oligopeptide, it is low to compensate for domestic ginseng
The blank of poly- peptide production, and the product yield that method using the present invention is prepared is high, the good, raciness of activity, has huge
Utilization foreground and market potential.
Figure of description:
Fig. 1：The molecular weight detection collection of illustrative plates of ginseng oligopeptide obtained by embodiment 1.
Specific implementation mode：
Embodiment 1：A kind of preparation method of ginseng oligopeptide, the preparation method include the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.8 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 5 hours at a temperature of 42 DEG C, then
PH is adjusted to 8.5, temperature and is adjusted to 28 DEG C, stirring and leaching 15 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.2 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.2, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 7.0 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
The dosage of step 3 neutral and alkali protease is 550-700U/g, and the temperature of enzymolysis is 45-50 DEG C for the first time, when enzymolysis
Between be 20-25min；The dosage of neutral proteinase is 200-300U/g in step 4, and the dosage of flavor protease is 700-900U/
The temperature of g, second of enzymolysis are 50-55 DEG C, enzymolysis time 60-120min.
Filtering in step 6 includes coarse filtration and refined filtration, and coarse filtration uses aperture to be carried out for the ceramic membrane of 400-1200nm, essence
Filter is carried out using the ceramic membrane of 50-200nm.
Embodiment 2：A kind of preparation method of ginseng oligopeptide, the preparation method include the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.5 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 8 hours at a temperature of 40 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 25 DEG C, stirring and leaching 12 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant liquid,
Take precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng egg
In vain；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.2, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 6.8 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
The dosage of step 3 neutral and alkali protease is 550-700U/g, and the temperature of enzymolysis is 45-50 DEG C for the first time, when enzymolysis
Between be 20-25min；The dosage of neutral proteinase is 200-300U/g in step 4, and the dosage of flavor protease is 700-900U/
The temperature of g, second of enzymolysis are 50-55 DEG C, enzymolysis time 60-120min.
Filtering in step 6 includes coarse filtration and refined filtration, and coarse filtration uses aperture to be carried out for the ceramic membrane of 400-1200nm, essence
Filter is carried out using the ceramic membrane of 50-200nm.
Embodiment 3：A kind of preparation method of ginseng oligopeptide, the preparation method include the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 5.0 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 4 hours at a temperature of 45 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 30 DEG C, stirring and leaching 17 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.5 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.5, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 7.2 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
The dosage of step 3 neutral and alkali protease is 550-700U/g, and the temperature of enzymolysis is 45-50 DEG C for the first time, when enzymolysis
Between be 20-25min；The dosage of neutral proteinase is 200-300U/g in step 4, and the dosage of flavor protease is 700-900U/
The temperature of g, second of enzymolysis are 50-55 DEG C, enzymolysis time 60-120min.
Filtering in step 6 includes coarse filtration and refined filtration, and coarse filtration uses aperture to be carried out for the ceramic membrane of 400-1200nm, essence
Filter is carried out using the ceramic membrane of 50-200nm.
Embodiment 4：Ginseng protein extraction experiment
Method 1（I.e. embodiment 1 the step of one to three）：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.8 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 5 hours at a temperature of 42 DEG C, then
PH is adjusted to 8.5, temperature and is adjusted to 28 DEG C, stirring and leaching 15 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.2 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen.
Method 2（Cellulase processing is omitted on the basis of method one）：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.8 of ginseng slurries；
The pH for the ginseng slurries that above-mentioned steps are prepared is adjusted to 8.5 by step 2, and temperature is adjusted to 28 DEG C, stirring and leaching
15 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant；
The pH of supernatant made from step 2 is adjusted to 4.2 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen.
Method 3（It is different from the technological parameter of embodiment 1）：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.8 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 5 hours at a temperature of 42 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 28 DEG C, stirring and leaching 15 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 3.8 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen.
The protein extraction yield of above method 1- methods 3 is respectively 18.35g albumen/kg ginsengs, 12.47g albumen/kg people
Ginseng, 13.59g albumen/kg ginsengs, it can be seen that, the method for embodiment 1 compares method 2（It is handled without cellulase）It improves
47.2%, compare method 3（It is different from the technological parameter of embodiment 1）Improve 35.0%, it can be seen that, people determined by the present invention
Ginseng albumen extraction process cellulase processing and technological parameter be the key that determine the present invention can it is successful and this
Two conditions ensure that the present invention can efficiently, economically obtain ginseng oligopeptide.
Embodiment 5：Ginseng oligopeptide flavor experiment
The oligomeric peptide product of Example 1-3 and commercially available soy oligosaccharides peptide product, are dissolved in distilled water, concentration of ordinary dissolution is
Ginseng oligopeptide solution is prepared in 100mg/10g, 500mg/10g, is evaluated by the food subjective appreciation mechanism of profession, special
It is not to be evaluated to its bitter taste, score is respectively embodiment 1（100mg/10g）9.2 points, embodiment 2（100mg/10g）
9.0 points, embodiment 3（100mg/10g）9.3 points, soybean oligopeptide（100mg/10g）8.7 point；Embodiment 1（500mg/10g）
9.0 points, embodiment 2（500mg/10g）8.9 points, embodiment 3（500mg/10g）9.1 points, soybean oligopeptide（500mg/10g）
8.2 points of it can be seen that the product of 1-3 of the embodiment of the present invention all without apparent bitter taste, and flavor evaluation result be superior to it is commercially available
Soy oligosaccharides peptide product.
Embodiment 6：By entrusting professional institution to be detected protein content in ginseng oligopeptide and oligopeptide, through inspection
It surveys, the protein content of the ginseng oligopeptide obtained by the embodiment of the present invention 1 is 90.35%（Butt）, the mass content of oligopeptide is
87.5%, the mass content that molecular weight is less than the peptide of 1000 dalton is 92.67%.
Claims (10)
1. a kind of ginseng oligopeptide, which is characterized in that the ginseng oligopeptide is prepared using the method for following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.5-5.0 of ginseng slurries；
Step 2 is added cellulase in the ginseng slurries obtained to step 1, is digested 3-8 hours at a temperature of 40-45 DEG C,
PH is then adjusted to 8.0-9.0, temperature is adjusted to 25-30 DEG C, stirring and leaching 12-17 hours, then centrifuged with 3500rpm rotating speeds
20min takes supernatant；
The pH of supernatant made from step 2 is adjusted to 4-4.5 by step 3, and stirring, 12000rpm centrifuge 10min, discard
Clear liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, that is, be people
Join albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid is adjusted to 8.2-8.5, alkali protease is then added thereto and carries out first time enzyme
Solution, obtains first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 6.8-7.2, neutral protein is then added thereto by step 5
Enzyme and flavor protease carry out second and digest, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
2. a kind of preparation method of ginseng oligopeptide, which is characterized in that the preparation method of the ginseng oligopeptide includes following step
Suddenly：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.5-5.0 of ginseng slurries；
Step 2 is added cellulase in the ginseng slurries obtained to step 1, is digested 3-8 hours at a temperature of 40-45 DEG C,
PH is then adjusted to 8.0-9.0, temperature is adjusted to 25-30 DEG C, stirring and leaching 12-17 hours, then centrifuged with 3500rpm rotating speeds
20min takes supernatant；
The pH of supernatant made from step 2 is adjusted to 4-4.5 by step 3, and stirring, 12000rpm centrifuge 10min, discard
Clear liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, that is, be people
Join albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid is adjusted to 8.2-8.5, alkali protease is then added thereto and carries out first time enzyme
Solution, obtains first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 6.8-7.2, neutral protein is then added thereto by step 5
Enzyme and flavor protease carry out second and digest, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
3. the preparation method of ginseng oligopeptide according to claim 2, it is preferable that the preparation method of the ginseng oligopeptide
Include the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.8 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 5 hours at a temperature of 42 DEG C, then
PH is adjusted to 8.5, temperature and is adjusted to 28 DEG C, stirring and leaching 15 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.2 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.2, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 7.0 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
4. the preparation method of ginseng oligopeptide according to claim 2, it is preferable that the preparation method of the ginseng oligopeptide
Include the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 4.5 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 8 hours at a temperature of 40 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 25 DEG C, stirring and leaching 12 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant liquid,
Take precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng egg
In vain；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.2, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 6.8 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
5. the preparation method of ginseng oligopeptide according to claim 2, it is preferable that the preparation method of the ginseng oligopeptide
Include the following steps：
Step 1 takes fresh ginseng, clean, drain after be cut into segment, with distilled water press 1:1 mass ratio mixing, then uses
Wall-breaking machine breaks into ginseng slurries, adjusts the pH to 5.0 of ginseng slurries；
Cellulase is added in the ginseng slurries obtained to step 1 in step 2, digests 4 hours at a temperature of 45 DEG C, then
PH is adjusted to 8.0, temperature and is adjusted to 30 DEG C, stirring and leaching 17 hours, then 20min is centrifuged with 3500rpm rotating speeds, take supernatant
Liquid；
The pH of supernatant made from step 2 is adjusted to 4.5 by step 3, and stirring, 12000rpm centrifuge 10min, discard supernatant
Liquid, takes precipitation, and will be again stirring for after precipitation mixes with distilled water, 12000rpm centrifuges 8min, collect precipitation, as ginseng
Albumen；
Step 4, by ginseng protein made from step 3 and distilled water according to 1:20 mass ratio mixing, is uniformly mixing to obtain people
Join protein liquid, and the pH of ginseng protein liquid be adjusted to 8.5, alkali protease is then added thereto and carries out first time enzymolysis,
Obtain first time enzymolysis liquid；
The pH of first time enzymolysis liquid made from step 4 is adjusted to 7.2 by step 5, then thereto be added neutral proteinase and
Flavor protease carries out second and digests, and after enzyme deactivation, the thick liquid of ginseng oligopeptide is made；
Step 6, after using aperture to be filtered to the thick liquid of ginseng oligopeptide made from step 5 for the ceramic membrane of 50-1200nm,
Filtrate is concentrated, is decolourized, is freeze-dried and obtains the ginseng oligopeptide.
6. the preparation method of ginseng oligopeptide according to claim 2, it is preferred that the dosage of step 3 neutral and alkali protease
For 550-700U/g, the temperature of enzymolysis is 45-50 DEG C for the first time, enzymolysis time 20-25min；Neutral proteinase in step 4
Dosage be 200-300U/g, the dosage of flavor protease is 700-900U/g, and the temperature of second of enzymolysis is 50-55 DEG C, enzyme
The solution time is 60-120min.
7. the preparation method of ginseng oligopeptide according to claim 2, it is preferable that the filtering in step 6 include coarse filtration and
Refined filtration, coarse filtration use aperture to be carried out for the ceramic membrane of 400-1200nm, and refined filtration is carried out using the ceramic membrane of 50-200nm.
8. a kind of food, which is characterized in that wherein comprising ginseng oligopeptide described in claim 1 or according to claim 2-
Ginseng oligopeptide obtained by the preparation method of 7 any one of them ginseng oligopeptides.
9. the preparation method of ginseng oligopeptide described in ginseng oligopeptide described in claim 1 or claim 2-7 any one
Application of the obtained ginseng oligopeptide in preparing food.
10. a kind of ginseng oligopeptide, which is characterized in that the ginseng oligopeptide is described in any one of claim 3-7
It is formed prepared by the preparation method of ginseng oligopeptide.
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| CN110839896A (en) * | 2019-12-26 | 2020-02-28 | 吉林大学 | Preparation method of ginseng instant food for improving type II diabetes symptoms |
| CN110846365A (en) * | 2019-11-28 | 2020-02-28 | 湖北瑞邦生物科技有限公司 | Mountain ginseng peptide, and preparation method and application thereof |
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