CN107828840A - Silkworm chrysalis active peptide, composition and preparation method - Google Patents
Silkworm chrysalis active peptide, composition and preparation method Download PDFInfo
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- 238000002360 preparation method Methods 0.000 title claims abstract description 89
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- 238000005238 degreasing Methods 0.000 description 8
- SHFGJEQAOUMGJM-UHFFFAOYSA-N dialuminum dipotassium disodium dioxosilane iron(3+) oxocalcium oxomagnesium oxygen(2-) Chemical compound [O--].[O--].[O--].[O--].[O--].[O--].[O--].[O--].[Na+].[Na+].[Al+3].[Al+3].[K+].[K+].[Fe+3].[Fe+3].O=[Mg].O=[Ca].O=[Si]=O SHFGJEQAOUMGJM-UHFFFAOYSA-N 0.000 description 8
- 235000019658 bitter taste Nutrition 0.000 description 7
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- 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 description 4
- QAOWNCQODCNURD-UHFFFAOYSA-N Sulfuric acid Chemical compound OS(O)(=O)=O QAOWNCQODCNURD-UHFFFAOYSA-N 0.000 description 4
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- 108010038983 glycyl-histidyl-lysine Proteins 0.000 description 4
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- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
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- 210000001367 artery Anatomy 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 235000019835 bromelain Nutrition 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 235000012000 cholesterol Nutrition 0.000 description 1
- 150000001841 cholesterols Chemical class 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 230000001079 digestive effect Effects 0.000 description 1
- 238000007598 dipping method Methods 0.000 description 1
- 238000004945 emulsification Methods 0.000 description 1
- 230000002255 enzymatic effect Effects 0.000 description 1
- 235000004626 essential fatty acids Nutrition 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 208000006454 hepatitis Diseases 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
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- 206010022437 insomnia Diseases 0.000 description 1
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- 229940055695 pancreatin Drugs 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000000049 pigment Substances 0.000 description 1
- 239000003531 protein hydrolysate Substances 0.000 description 1
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- 238000011084 recovery Methods 0.000 description 1
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- 230000035882 stress Effects 0.000 description 1
- -1 trypsase Proteins 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 238000010792 warming Methods 0.000 description 1
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES, NOT OTHERWISE PROVIDED FOR; PREPARATION OR TREATMENT THEREOF
- 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
-
- 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
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- Proteomics, Peptides & Aminoacids (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
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Abstract
The invention provides a kind of preparation method of silkworm chrysalis active peptide and silkworm chrysalis active peptide.The preparation method comprises the following steps：A, dried silkworm chrysalis meal is added into water, adjust pH to 8~9, immersion 18~24 hours, separation of solid and liquid, supernatant regulation pH to 4~6 is precipitated, separation of solid and liquid, solid portion is added into water, 15~30min is heated at 75~100 DEG C, then carries out homogeneous, obtained suspension is adjusted into pH to 5~10, enzyme preparation is added and is digested；The enzyme preparation is made up of flavor protease, papain and the neutral proteinase that weight ratio is 1~2: 1~2: 1~2, and hydrolysis temperature is 40~65 DEG C, and enzymolysis time is 1.5~8h, obtains enzymolysis liquid, and by enzymolysis liquid destroy the enzyme treatment, silkworm chrysalis active peptide powder is made.The preparation method simple production process of the present invention, and obtained silkworm chrysalis active peptide purity is high, small peptide content is high.
Description
Technical field
The invention belongs to biological technical field, and in particular to a kind of silkworm chrysalis active peptide, composition and preparation method.
Background technology
Fresh pupa yield is up to more than 300,000 tons every year in China.Protein content is up to 52~65% in dry silkworm chrysalis, and amino acid
A wide selection of colours and designs, proportioning equalize, it is necessary to which amino acid content enriches, and is a kind of high-quality protein.Contain essential fatty acid, energy in addition
Enough dissolving cholesterols, have the effect of good to angiocarpy, artery sclerosis, hypertension, chronic hepatitis etc..But pupa albumen is water-soluble
Difference, containing smell substance and pigment, particularly after water-swellable, peculiar smell and color and luster are even more serious, hinder it in the food industry
Application.
With the application of protein engineering, grinding for biologically active peptide and its protolysate is produced using zymolysis technique
Study carefully and have become focus with product development.Has the laboratory that document report enzymolysis silkworm chrysalis protein in part produces polypeptide at present
Technology.
The Chinese patent application of Application No. 201611197961.1 discloses a kind of preparation method of pupa albumen Gly-His-Lys,
Silkworm chrysalis is subjected to degreasing operation first, obtains the pupa albumen liquid of degreasing；The pupa albumen liquid of the degreasing is molten with 10% sulfuric acid
Liquid or hydrochloric acid solution regulation pH are 4.0~4.2, stand, precipitation and centrifugal separation, obtain silkworm chrysalis crude separation albumen；By the crude separation
Albumen pH, 4.0~4.2 pure water washing desalination, stand again, centrifugation and desalting processing, obtain the silkworm chrysalis separation of desalination
Albumen；Moisture is added to dissipate the silkworm separated protein of the desalination, it is 9.0~9.5 to adjust pH with sodium hydroxide solution, is warming up to 50~55
DEG C, 40~60min of trypsin digestion is firstly added, adds 2~3h of papain enzymolysis, obtains pupa albumen enzymolysis
Liquid；Then through decolourizing and being dried to obtain pupa albumen Gly-His-Lys.For this method through degreasing, regulation pH is acidity, separates crude protein, then
Through desalination, successively digested with trypsase and papain, obtain Silkworm pupa protein.
Application No. 201210062890.X Chinese patent application discloses a kind of silkworm chrysalis method of comprehensive utilization, and it is first
Degreasing extraction silkworm chrysalis oil after dried silkworm chrysalis meal is made, extracted first with alkaline solution, obtain pupa albumen solution, then adjust pH to
4.5, stand, collect precipitation, add pH6.0 buffer solution and hydrogen peroxide, heating stirring, boil, re-adjustments pH, standing, mistake
Filter, the step of precipitation is collected, after drying, add buffer solution, digested using proteolytic enzyme, obtain silkworm chrysalis polypeptide powder.Its
Middle proteolytic enzyme includes pepsin, pancreatin, trypsase, papain, bromelain.
The preparation method of above-mentioned Silkworm pupa protein needs to carry out cumbersome degreasing, de-salting operation, complex process.It is above-mentioned existing
The small peptide content of Silkworm pupa protein made from technology is relatively low, and activity is low；And taste is partially bitter, mouthfeel is bad.Furthermore above-mentioned existing skill
Pupa albumen powder purity is relatively low made from art.
The content of the invention
An object of the present disclosure is to provide a kind of preparation method of silkworm chrysalis active peptide, and this method technique is simple, and is made
Silkworm chrysalis activity peptide activity it is high, no bitter taste.
Another object of the present invention is to provide using silkworm chrysalis active peptide made from above-mentioned preparation method, the silkworm chrysalis active peptide
Content of peptides is high, no bitter taste.
It is still another object of the present invention to provide a kind of composition containing silkworm chrysalis active peptide.
A further object of the present invention is the preparation method for providing the composition containing silkworm chrysalis active peptide.
The preparation method of the silkworm chrysalis active peptide of the present invention, comprises the following steps：
A, dried silkworm chrysalis meal is added into water, adjusts pH to 8~9, soaked 18~24 hours, separation of solid and liquid, take supernatant；
B, supernatant regulation pH to 4~6 is precipitated, separation of solid and liquid, takes solid portion；
C, the solid portion is added into water, 15~30min is heated at 75~100 DEG C, homogeneous is then carried out, is mixed
Suspension；
D, the suspension is adjusted into pH to 5~10, adds enzyme preparation and digested；The enzyme preparation is by flavor albumen
Enzyme, papain and neutral proteinase composition, and the weight ratio of the flavor protease, papain and neutral proteinase
For 1~2: 1~2: 1~2, hydrolysis temperature is 40~65 DEG C, and enzymolysis time is 1.5~8h, obtains enzymolysis liquid；
E, by the enzymolysis liquid destroy the enzyme treatment, silkworm chrysalis active peptide powder is then made.
Preparation in accordance with the present invention, it is preferable that in step B, the supernatant is adjusted by pH to 4~6 using weak acid；
The weak acid is selected from least one of acetic acid, lactic acid, citric acid.
Preparation in accordance with the present invention, it is preferable that in step E, the temperature of the destroy the enzyme treatment is 90~125 DEG C, when
Between be 0.5~3h.
Preparation in accordance with the present invention, it is preferable that in step E, the enzymolysis liquid after the destroy the enzyme treatment is concentrated, homogeneous
Emulsification, is then spray-dried, obtains silkworm chrysalis active peptide powder；The spray drying is carried out at 80~105 DEG C.
Preparation in accordance with the present invention, it is preferable that in step E, the enzymolysis liquid after the destroy the enzyme treatment is centrifuged,
Centrifugate filters through plate and frame filter press first, then through ceramic membrane filter, obtains active peptide permeate；The active peptide filtrate is dense
Contracting, spray drying, so as to obtain silkworm chrysalis active peptide powder；The spray drying is carried out at 80~105 DEG C；The plate and frame filter press
It is interior containing combination filtration adjuvant, it is described combination filtration adjuvant by activated carbon, perlite and diatomite according to weight ratio 6~7: 1~2
: 2~3 compositions.
The present invention also provides a kind of silkworm chrysalis active peptide, and the silkworm chrysalis active peptide is prepared using the above method.
The present invention also provides a kind of composition of active peptide containing silkworm chrysalis, and it is by the parts by weight of silkworm chrysalis active peptide 90~98, mulberries 1
~5 parts by weight and the parts by weight of spina date seed 1~5 are made.
The present invention also provides a kind of preparation method of the composition of the active peptide containing silkworm chrysalis, and the preparation method includes：
(1) silkworm chrysalis active peptide powder is prepared using the above method；(2) mulberries and spina date seed are prepared as mulberries powder and spiny jujuba seed powder respectively；
(3) silkworm chrysalis active peptide powder, mulberries powder and the spiny jujuba seed powder are well mixed, obtain the composition；Or
The preparation method includes：(1) silkworm chrysalis active peptide powder is prepared using the above method；(2) mulberries and spina date seed are mixed
By the use of water as solvent extraction after conjunction, extract solution concentration, it dry, pulverize, obtain medicinal substances extract；(3) by the silkworm chrysalis active peptide
Powder and medicinal substances extract are well mixed, and obtain the composition.
The present invention also provides the composition of another active peptide containing silkworm chrysalis, its by the parts by weight of dried silkworm chrysalis meal 90~98, mulberries 1~
5 parts by weight and the parts by weight of spina date seed 1~5 are made.
The present invention also provides a kind of preparation method of the composition of the active peptide containing silkworm chrysalis, including：
A ', dried silkworm chrysalis meal is added to water, adjust pH to 8~9, soaked 18~24 hours, separation of solid and liquid, take supernatant；
B ', by the supernatant regulation pH to 4~6 precipitated, separation of solid and liquid, take solid portion；
C ', by the solid portion add water, at 75~100 DEG C heat 15~30min, then carry out homogeneous, obtain
Suspension；
D ', the suspension is adjusted to pH to 5~10, addition enzyme preparation is digested；The enzyme preparation is by flavor albumen
Enzyme, papain and neutral proteinase composition, and the weight ratio of the flavor protease, papain and neutral proteinase
For 1~2: 1~2: 1~2；Hydrolysis temperature is 40-65 DEG C, enzymolysis time 1.5-8h, obtains enzymolysis liquid；
D1 ', the mulberries and spina date seed added in the enzymolysis liquid, heating and refluxing extraction, filtering, obtain mixed liquor；
E ', the composition that the mixed liquor is made to the active peptide containing silkworm chrysalis.
The preparation method of the present invention eliminates degreasing and desalting steps, and technique is simple, and cost is low.Made from the inventive method
Silkworm chrysalis active peptide small peptide content is higher, and purity is high.The composition of the active peptide containing silkworm chrysalis of the present invention can improve human immunity
Function, anti-aging, and there is liver-kidney tonifying, the effect of antitoxic heart-soothing and sedative.
Embodiment
With reference to specific embodiment, the present invention is further illustrated, but protection scope of the present invention is not limited to
This.
<The preparation method and silkworm chrysalis active peptide of silkworm chrysalis active peptide>
The preparation method of the silkworm chrysalis active peptide of the present invention, comprises the following steps：
A, dried silkworm chrysalis meal is added into water, adjusts pH to 8~9, soaked 18~24 hours, separation of solid and liquid, take supernatant；
B, supernatant regulation pH to 4~6 is precipitated, separation of solid and liquid, takes solid portion；
C, the solid portion is added into water, 15~30min is heated at 75~100 DEG C, homogeneous is then carried out, is mixed
Suspension；
D, the suspension is adjusted into pH to 5~10, adds enzyme preparation and digested；The enzyme preparation is by flavor albumen
Enzyme, papain and neutral proteinase composition, and the weight ratio of the flavor protease, papain and neutral proteinase
For 1~2: 1~2: 1~2；Hydrolysis temperature is 40~65 DEG C, and enzymolysis time is 1.5~8h, obtains enzymolysis liquid；
E, by the enzymolysis liquid destroy the enzyme treatment, silkworm chrysalis active peptide powder is then made.
Preparation in accordance with the present invention, in step A, the particle diameter of the dried silkworm chrysalis meal is 2~75 μm.The dried silkworm chrysalis meal can be with
By the way that dry silkworm chrysalis is carried out into ultramicro grinding, then sieving obtains, such as crosses 200~600 mesh sieves.Using the silkworm of the particle diameter of the present invention
Pupa powder, enzymatic hydrolyzation can be effectively improved in the enzymolysis step in later stage, make full use of the protein ingredient in silkworm chrysalis.The immersion is excellent
Elect as and soaked under agitation, so that the composition in dried silkworm chrysalis meal is fully dissolved out.In step A, amount of water is dried silkworm chrysalis meal weight
3~10 times, preferably 3~5 times.Preferably, after the completion of process is soaked, mechanical agitation is carried out, so that liquid is uniform, just
In centrifugation.
In step A, the solid-liquid separating method can be filtering, centrifugation etc., and be preferably to centrifuge.In step A, leaching
Bubble process can be heated or carried out at normal temperatures, and preferably be carried out at normal temperatures.Generally, need to be heated during dipping by lye
Processing；And inventor has found, the composition in dried silkworm chrysalis meal can be fully dissolved out by immersion at normal temperatures, so as to avoid carrying out heating production
Raw additional energy, and more convenient operation.
Preparation in accordance with the present invention, in step B, the supernatant is adjusted by pH to 4~6 using weak acid.It is described weak
Acid can be selected from least one of acetic acid, lactic acid, citric acid.In existing silkworm chrysalis active peptide extracting method, it is contemplated that regulation
Efficiency, the step for the strong acid for adjusting pH value of generally use to acidity, the strong acid is, for example, hydrochloric acid, sulfuric acid etc., and less consideration
Influence of the property of acid for product；Inventor has found that strong acid is larger for protein component destruction, to the silkworm chrysalis finally given
Active peptide Product Activity has a negative impact.And weak acid for adjusting pH is used, protein precipitation is come out, reduces and protein is sought
The destruction of material is supported, and the silkworm chrysalis active peptide Product Activity finally given is more preferable.In step B, it is preferable that the sedimentation time is
8~12h, so as to ensure that the protein component in supernatant is fully precipitated out under above-mentioned pH.Preferably, process is precipitated to complete
Mechanical agitation is carried out afterwards to carry out under agitation, so that liquid is uniform, is easy to centrifuge.
Preparation in accordance with the present invention, in step C, make protein denaturation using heating means.In described fire-bar
It can realize that the protein component in silkworm chrysalis is denatured completely under part, and be advantageous to fully be digested in subsequent step, simultaneously
And avoid pollution of the chemical reagent to product.Homogeneous is carried out after heat distortion, further increases enzymolysis efficiency.
Preparation in accordance with the present invention, in step D, the suspension is adjusted to pH5~10, preferably pH7~9,
More preferably pH7~8.In the present invention, pH is adjusted to above-mentioned alkalescent, be more beneficial for the present invention enzyme preparation digested.
In the present invention, enzyme preparation is 1~2: 1~2: 1~2 according to weight ratio by flavor protease, papain and neutral proteinase
Composition.According to the preferred embodiment of the present invention, the weight ratio of flavor protease, papain and neutral proteinase is 1.3
~1.8: 1.3~1.8: 1~1.5.According to one preferred embodiment of the invention, flavor protease, papain and in
Property protease weight ratio be 1.5: 1.5: 1.The enzymolysis process of the present invention disposably adds above-mentioned enzyme preparation, and in above-mentioned pH bars
Digested under part, more preferably hydrolysis result can be obtained.Enzymolysis of the enzyme preparation of the present invention for the protein component in silkworm chrysalis
Efficiency high, hence it is evident that it is alone higher than every kind of enzyme, and it is higher than existing silkworm chrysalis active peptide enzyme.When flavor protease, papain
When weight ratio with neutral proteinase is 1.5: 1.5: 1, enzymolysis efficiency highest.Preparation in accordance with the present invention, the enzyme system
The addition of agent can be 0.1~2wt%, the preferably 0.3~1.5wt% of solid content weight in the suspension, more preferably
For 0.4~1.2wt%.In the present invention, hydrolysis temperature is 40~65 DEG C, and enzymolysis time is 1.5~8h；Preferably, hydrolysis temperature
For 45~60 DEG C, enzymolysis time is 2~6h.The addition of above-mentioned enzyme preparation, enzymolysis time and at a temperature of, pupa albumen can be real
Existing effect enzymolysis.
Preparation in accordance with the present invention, in step E, the temperature of the destroy the enzyme treatment is 90~125 DEG C, the time 0.5
~3h.In step E, silkworm chrysalis active peptide powder can be made using conventional method in the enzymolysis liquid after destroy the enzyme treatment.
According to an embodiment of the invention, the enzymolysis liquid after the destroy the enzyme treatment is concentrated, emulsifying, then sprayed
Mist is dried, and obtains silkworm chrysalis active peptide powder, the silkworm chrysalis active peptide powder is total nutrient silkworm chrysalis active peptide.The total nutrient silkworm chrysalis active peptide produces
In product, peptide content >=40wt%, wherein.Preferably, the spray-drying operation is carried out at 80~105 DEG C, so as to effectively keep
The activity of silkworm chrysalis active peptide.
According to another preferred embodiment of the present invention, the enzymolysis liquid after the destroy the enzyme treatment is centrifuged, centrifugate
Filtered first through plate and frame filter press, then through ceramic membrane filter, obtain active peptide permeate；The active peptide filtrate is concentrated, spray
Mist is dried, and so as to obtain silkworm chrysalis active peptide powder, the silkworm chrysalis active peptide powder is refined silkworm chrysalis active peptide powder.
In the present invention, containing combination filtration adjuvant in the plate and frame filter press, the combination filtration adjuvant is by activated carbon, treasure
Zhu Yan and diatomite composition, and activated carbon, perlite and diatomaceous weight ratio 6~7: 1~2: 2~3.According to the one of the present invention
Individual preferred embodiment, it is described to combine activated carbon, perlite and diatomaceous weight ratio 7: 1.5: 2 in filtration adjuvant.According to
Another preferred embodiment of the present invention, it is described to combine activated carbon, perlite and diatomaceous weight ratio in filtration adjuvant
6.7∶1.8∶2.Inventor has found that silkworm chrysalis active peptide prepared by existing method carries peculiar smell, and has bitter taste, poor taste；It is a variety of
Filtration adjuvant is alone to be difficult to simultaneously remove the two.The present invention filtration adjuvant can effectively remove silkworm chrysalis per se with it is different
Taste and bitter taste, and after the filtration adjuvant processing by the present invention, grease-like composition is removed in enzymolysis liquid, thus need not be entered early stage
The cumbersome degreasing operation of row, simplifies technological process.In the present invention, the aperture of the ceramic membrane can be 20~500nm, more excellent
Elect 20~300mm as.Using such ceramic membrane aperture, further refining for enzymolysis liquid can be realized.
After filtration treatment by the present invention, peptide content >=85wt% in obtained refined silkworm chrysalis active peptide powder.
Generally, the spray-drying operation of peptide is in 120~140 DEG C of progress, it is considered that such drying temperature is for peptide
Activity influence is little.However, it is found by the inventors that the temperature of existing spray drying significantly affects the activity of peptide.And in the present invention,
Preferably, the spray-drying operation is in 80~105 DEG C of progress, so as to effectively keep the activity of silkworm chrysalis active peptide.
<Composition of the active peptide containing silkworm chrysalis and preparation method thereof>
The composition of the active peptide containing silkworm chrysalis of the present invention can be by the parts by weight of silkworm chrysalis active peptide 90~98, the weight of mulberries 1~5
Part and the parts by weight of spina date seed 1~5 are made；Or
The present invention the active peptide containing silkworm chrysalis composition can by the parts by weight of dried silkworm chrysalis meal 90~98, the parts by weight of mulberries 1~5 and
The parts by weight of spina date seed 1~5 are made.
In the present invention, silkworm chrysalis active peptide can use what the above-mentioned preparation method of the present invention was prepared.Silkworm chrysalis active peptide
Dissolubility and digestive utilization ratio are high, no dependence, leucocyte level in human body can be effectively improved, so as to improve human immunity work(
Can, delaying human body function aging.Mulberries have liver-kidney tonifying, the effect of nourishing yin and nourishing blood, are lost for treating kidney deficiency and liver and deficiency of blood essence
Caused many illnesss.Spina date seed then has nourishing heart tonifying liver, the effect of antitoxic heart-soothing and sedative, for treating restlessness of asrhenia type and insomnia, horrified to much dream.This hair
Bright silkworm chrysalis active peptide coordinates a small amount of mulberries and spina date seed, is improving immune function of human body, on the basis of delaying human body function aging,
With liver-kidney tonifying, the effect of antitoxic heart-soothing and sedative, especially suitable the elderly takes.
The preparation method of the composition of the active peptide containing silkworm chrysalis of the present invention, can be obtained using a variety of preparation methods.
According to an embodiment of the invention, the preparation method of the composition is：(1) silkworm chrysalis active peptide powder is prepared；
(2) mulberries and spina date seed are prepared as mulberries powder and spiny jujuba seed powder respectively；(3) by silkworm chrysalis active peptide powder, mulberries powder and the acid
Jujube kernel powder is well mixed, and obtains the composition of the present invention.The silkworm chrysalis active peptide powder is preferably prepared using above-mentioned preparation method
Obtain.The mulberries powder is preferably that mulberries progress ultramicro grinding is made.The spiny jujuba seed powder is preferably to be surpassed spina date seed
Crushing of Ultrafine is made.Wherein, step (1 ') is made using above-mentioned " preparation method of silkworm chrysalis active peptide ", is repeated no more here.
According to another implementation of the invention, the preparation method of the composition is：(1 ') prepares silkworm chrysalis active peptide
Powder；Mulberries and spina date seed by the use of water as solvent extraction, extract solution concentration, dry, pulverize, obtain medicinal substances extract by (2 ')；
(3 ') mix the silkworm chrysalis active peptide powder and medicinal substances extract, obtain the composition of the present invention.Wherein, in step (1 ') use
State " preparation method of silkworm chrysalis active peptide " to be made, repeat no more here.
The composition of the active peptide containing silkworm chrysalis of the present invention can also be by the parts by weight of dried silkworm chrysalis meal 90~98, the parts by weight of mulberries 1~5
It is made with the parts by weight of spina date seed 1~5.
Preferably, the preparation method of the composition of the peptide containing silkworm chrysalis is：
A ', dried silkworm chrysalis meal is added to water, adjust pH to 8~9, soaked 18~24 hours, separation of solid and liquid, take supernatant；
B ', by the supernatant regulation pH to 4~6 precipitated, separation of solid and liquid, take solid portion；
C ', by the solid portion add water, at 75~100 DEG C heat 15~30min, then carry out homogeneous, obtain
Suspension；
D ', the suspension is adjusted to pH to 5~10, addition enzyme preparation is digested；The enzyme preparation is by flavor albumen
Enzyme, papain and neutral proteinase composition, and the weight ratio of the flavor protease, papain and neutral proteinase
For 1~2: 1~2: 1~2；Hydrolysis temperature is 40-65 DEG C, enzymolysis time 1.5-8h, obtains enzymolysis liquid；
D1 ', the mulberries and spina date seed added in the enzymolysis liquid, heating and refluxing extraction, filtering, obtain mixed liquor；
E ', the composition that the mixed liquor is made to the active peptide containing silkworm chrysalis.
Wherein, step A '~D ' concrete operations and parameter respectively with A~D in above-mentioned " preparation method of silkworm chrysalis active peptide "
It is identical, repeat no more here.
In step D1 ', it is preferable that the mulberries and spina date seed add in the enzymolysis liquid in powder form, so as to improve
Recovery rate.The heating and refluxing extraction time is 1~3 hour, preferably 1~2 hour.The filtering can use customary filtration methods,
So as to filter off the dregs of a decoction, mixed liquor is obtained.
According to one preferred embodiment of the invention, in step E ', the mixed liquor that step D1 ' is obtained can be carried out dense
Contracting, emulsifying, is then spray-dried, obtains the composition of the active peptide containing silkworm chrysalis of full nutrition.Preferably, the spray drying
Operation is in 80~105 DEG C of progress, so as to effectively keep the activity of silkworm chrysalis active peptide.The group of the active peptide containing silkworm chrysalis of the full nutrition
In compound, peptide content >=30wt%.
According to presently preferred embodiment, in step E ', the mixed liquor is centrifuged, centrifugate is first
First filtered through plate and frame filter press, then through ceramic membrane filter, filtrate concentration, spray drying, it is refined containing silkworm chrysalis activity so as to obtain
The composition of peptide.Preferably, the spray-drying operation is in 80~105 DEG C of progress, so as to effectively keep the work of silkworm chrysalis active peptide
Property.
It is described to combine filtration adjuvant as described above containing combination filtration adjuvant in the plate and frame filter press in the present invention,
Here repeat no more.The present invention filtration adjuvant except remove silkworm chrysalis per se with peculiar smell and bitter taste, remove grease-like composition,
It also removes the non-solubility composition in the mulberries and spina date seed in mixed liquor.In the present invention, the aperture of the ceramic membrane can be with
For 20~500nm, more preferably 20~300mm.Using such ceramic membrane aperture, the further essence of mixed liquor can be realized
System.Preferably, the spray-drying operation is in 80~105 DEG C of progress, so as to effectively keep the activity of silkworm chrysalis active peptide.
After filtration treatment by the present invention, the composition of the refined active peptide containing silkworm chrysalis, peptide content >=85wt% are obtained.
Embodiments of the invention are described in detail below, but protection scope of the present invention is not limited.
Embodiment 1
The preparation method of total nutrient silkworm chrysalis active peptide, comprises the following steps：
A, dry silkworm chrysalis is subjected to ultramicro grinding, crosses 400 mesh sieves, obtain dried silkworm chrysalis meal；Dried silkworm chrysalis meal is added into 4 times of dried silkworm chrysalis meal weight
The water of amount, mixing, adjusted with sodium hydroxide solution to pH8, soak 20h；Mechanical agitation, centrifugation, takes supernatant；
B, the supernatant is adjusted to pH5, precipitating proteins, mechanical agitation with acetum, centrifugation, takes solid part
Point；
C, the solid portion is added into water, 20min is heated at 90 DEG C, homogeneous is then carried out, obtains suspension；
D, the suspension is adjusted into pH to 8, adds enzyme preparation and digested, obtain enzymolysis liquid；The enzyme preparation is by flavor
Protease, papain and neutral proteinase form according to weight ratio 1.5: 1.5: 1；The dosage of enzyme preparation is solid in suspension
The 0.5wt% of shape thing weight；Hydrolysis temperature is 50 DEG C, enzymolysis time 2h；
E, the enzymolysis liquid is subjected to destroy the enzyme treatment, enzyme-removal temperature is 90 DEG C, time 1h；Enzymolysis solution after enzyme deactivation is dense
Contracting, emulsifying, in 85 DEG C of spray drying in middle low-temperature spray drying tower, obtain total nutrient silkworm chrysalis active peptide powder.
Embodiment 2
The preparation method of total nutrient silkworm chrysalis active peptide, comprises the following steps：
A, dry silkworm chrysalis is subjected to ultramicro grinding, crosses 400 mesh sieves, obtain dried silkworm chrysalis meal；Dried silkworm chrysalis meal is added into dried silkworm chrysalis meal weight 3.5
The water of amount again, mixing, adjusted with sodium hydroxide solution to pH9, soak 20h；Mechanical agitation, centrifugation, takes supernatant；
B, the supernatant is adjusted to pH6, precipitating proteins, mechanical agitation with acetum, centrifugation, takes solid part
Point；
C, the solid portion is added into water, 15min is heated at 90 DEG C, homogeneous is then carried out, obtains suspension；
D, the suspension is adjusted into pH to 7.5, adds enzyme preparation and digested, obtain enzymolysis liquid；The enzyme preparation is by wind
Taste protease, papain and neutral proteinase form according to weight ratio 1.4: 1.6: 1.2；The dosage of enzyme preparation is suspension
The 0.4wt% of middle solid content weight；Hydrolysis temperature is 50 DEG C, enzymolysis time 2h；
E, the enzymolysis liquid is subjected to destroy the enzyme treatment, enzyme-removal temperature is 95 DEG C, time 1h；Enzymolysis solution after enzyme deactivation is dense
Contracting, emulsifying, in 100 DEG C of spray drying in middle low-temperature spray drying tower, obtain total nutrient silkworm chrysalis active peptide powder.
Embodiment 3
The preparation method of refined silkworm chrysalis active peptide, comprises the following steps：
A, dry silkworm chrysalis is subjected to ultramicro grinding, crosses 400 mesh sieves, obtain dried silkworm chrysalis meal；Dried silkworm chrysalis meal is added into 4 times of dried silkworm chrysalis meal weight
The water of amount, mixing, adjusted with sodium hydroxide solution to pH8, soak 20h；Mechanical agitation, centrifugation, takes supernatant；
B, the supernatant is adjusted to pH5, precipitating proteins, mechanical agitation with acetum, centrifugation, takes solid part
Point；
C, the solid portion is added into water, 20min is heated at 90 DEG C, homogeneous is then carried out, obtains suspension；
D, the suspension is adjusted into pH to 8, adds enzyme preparation and digested, obtain enzymolysis liquid；The enzyme preparation is by flavor
Protease, papain and neutral proteinase are according to 1.5: 1.5: 1 compositions；The dosage of enzyme preparation is solid content weight in suspension
The 0.5wt% of amount；Hydrolysis temperature is 50 DEG C, enzymolysis time 2h；
E, the enzymolysis liquid is subjected to destroy the enzyme treatment, enzyme-removal temperature is 90 DEG C, time 1h；By enzymolysis solution after enzyme deactivation from
The heart, centrifugate filter through plate and frame filter press, contain filtration adjuvant in plate and frame filter press, filtration adjuvant by activated carbon, perlite and
Diatomite is according to weight than 7: 1.5: 2 compositions；Via hole diameter is 400nm ceramic membrane filter again, obtains active peptide permeate；Should
Active peptide permeate concentrates, and in 85 DEG C of spray drying in middle low-temperature spray drying tower, obtains refined silkworm chrysalis active peptide powder.
Embodiment 4
The preparation method of refined silkworm chrysalis active peptide, comprises the following steps：
A, dry silkworm chrysalis is subjected to ultramicro grinding, crosses 400 mesh sieves, obtain dried silkworm chrysalis meal；Dried silkworm chrysalis meal is added into 4 times of dried silkworm chrysalis meal weight
The water of amount, mixing, adjusted with sodium hydroxide solution to pH8, soak 20h；Mechanical agitation, centrifugation, takes supernatant；
B, the supernatant is adjusted to pH5, precipitating proteins, mechanical agitation with acetum, centrifugation, takes solid part
Point；
C, the solid portion is added into water, 20min is heated at 90 DEG C, homogeneous is then carried out, obtains suspension；
D, the suspension is adjusted into pH to 8, adds enzyme preparation and digested, obtain enzymolysis liquid；The enzyme preparation is by flavor
Protease, papain and neutral proteinase are according to 1.6: 1.6: 1.2 compositions；The dosage of enzyme preparation is solid content in suspension
The 0.7wt% of weight；Hydrolysis temperature is 50 DEG C, enzymolysis time 2h；
E, the enzymolysis liquid is subjected to destroy the enzyme treatment, enzyme-removal temperature is 90 DEG C, time 1h；By enzymolysis solution after enzyme deactivation from
The heart, centrifugate filter through plate and frame filter press, contain filtration adjuvant in plate and frame filter press, filtration adjuvant by activated carbon, perlite and
Diatomite is according to weight than 6.7: 1.8: 2 compositions；Via hole diameter is 400nm ceramic membrane filter again, obtains active peptide permeate；Will
The active peptide permeate concentrates, and in 85 DEG C of spray drying in middle low-temperature spray drying tower, obtains refined silkworm chrysalis active peptide powder.
Embodiment 5
Mulberries powder and spiny jujuba seed powder is made in mulberries and spina date seed ultramicro grinding respectively.By the silkworm chrysalis active peptide of embodiment 1
4 parts of the parts by weight of powder 92, the parts by weight of mulberries powder 4 and spiny jujuba seed powder are well mixed, obtain the combination of the active peptide containing silkworm chrysalis of full nutrition
Thing.
Embodiment 6
The parts by weight of mulberries 3 and the parts by weight of spina date seed 3 are mixed, water is added as solvent heating and refluxing extraction 3 times, adds every time
Water is followed successively by the 8 times of amounts, 6 times of amounts, 6 times of amounts of mulberries and spina date seed gross weight, each extraction time be followed successively by 1.5 hours, it is 1 small
When, 1 hour, extract solution concentration, dry, pulverize, obtain medicinal substances extract.By the parts by weight of silkworm chrysalis active peptide powder 94 of embodiment 1
It is well mixed with above-mentioned medicinal substances extract, obtains the composition of the active peptide containing silkworm chrysalis of full nutrition.
Embodiment 7
Mulberries powder and spiny jujuba seed powder is made in mulberries and spina date seed ultramicro grinding respectively.By the silkworm chrysalis active peptide of embodiment 3
4 parts of the parts by weight of powder 92, the parts by weight of mulberries powder 4 and spiny jujuba seed powder are well mixed, obtain the group of the active peptide containing silkworm chrysalis of the present embodiment
Compound.
Embodiment 8
A ', dry silkworm chrysalis is carried out to ultramicro grinding, excessively 400 mesh sieves, obtain the parts by weight of dried silkworm chrysalis meal 94；Dried silkworm chrysalis meal is added into silkworm chrysalis
The water of 4 times of amounts of powder weight, mixing, adjusted with sodium hydroxide solution to pH 8, soak 20h；Mechanical agitation, centrifugation, takes supernatant；
B ', the supernatant adjusted to pH5, precipitating proteins, mechanical agitation with acetum, centrifugation, take solid part
Point；
C ', by the solid portion add water, 20min is heated at 90 DEG C, homogeneous is then carried out, obtains suspension；
D ', the suspension is adjusted to pH to 8, addition enzyme preparation is digested, and obtains enzymolysis liquid；The enzyme preparation is by flavor
Protease, papain and neutral proteinase are according to 1.5: 1.5: 1 compositions；The dosage of enzyme preparation is solid content weight in suspension
The 0.5wt% of amount；Hydrolysis temperature is 50 DEG C, enzymolysis time 2h；
D1 ', the parts by weight of mulberries 3 and the parts by weight of spina date seed 3 are added in the enzymolysis liquid, heating and refluxing extraction, during extraction
Between be 2h, filtering, obtain mixed liquor；
E ', the mixed liquor being centrifuged, centrifugate is filtered through plate and frame filter press, and filtration adjuvant is contained in plate and frame filter press,
Filtration adjuvant is made up of activated carbon, perlite and diatomite according to weight ratio 7: 1.5: 2；Via hole diameter is 400nm ceramic membrane again
Filtering, filtrate concentration, in 85 DEG C of spray drying in middle low-temperature spray drying tower, obtain the composition of the active peptide containing silkworm chrysalis.
Comparative example 1
Except the enzyme preparation of step D in embodiment 3 is replaced with by flavor protease, papain and neutral proteinase
It is same as Example 3 according to 3: 3: 1 compositions, other steps.
Comparative example 2
Except the acetum of step B in embodiment 1 is replaced with into hydrochloric acid solution, other steps are identical with documents 1.
Comparative example 3
Activated carbon except the filtration adjuvant of step E in embodiment 3 to be replaced with to equivalent weight, other steps and embodiment
3 is identical.
Comparative example 4
Perlite except the filtration adjuvant of step E in embodiment 3 to be replaced with to equivalent weight, other steps and embodiment
3 is identical.
Comparative example 5
Diatomite except the filtration adjuvant of step E in embodiment 3 to be replaced with to equivalent weight, other steps and embodiment
3 is identical.
Comparative example 6
Except the spray drying temperature of step E in embodiment 3 is arranged into 135 DEG C, other steps are same as Example 3.
Comparative example 7
Method according to the Chinese patent application specification embodiment 1 of Application No. 201611197961.1 prepares silkworm chrysalis work
Property Gly-His-Lys.
Comparative example 8
Method according to Application No. 201210062890.X Chinese patent application specification embodiment 1 prepares silkworm chrysalis work
Property Gly-His-Lys.
Peptide content the results are shown in Table 1 in silkworm chrysalis active peptide in measure embodiment 1~4 and comparative example 1~2,6~8.
The content assaying method of peptide is：Carried out according to the assay method of peptide content in the soy peptide powders of GB/T 22492.
Bitter taste estimate method is：Take 1g silkworm chrysalises active peptide to add in 10ml warm water to stir, extension rate is calculated as hardship
Taste value, the average bitterness value of 20 people is calculated, the results are shown in Table 2.
Fishy smell evaluation method is：Evaluation method is：0- is without fishy smell；1- slightly has fishy smell；2- fishy smell is weaker；3- has fishy smell；4-
Fishy smell is general；5- fishy smell is laid particular stress on；6- fishy smell is heavier；The average fishy smell value of the 7- fishy smell very people of re-computation 20, the results are shown in Table 2.
Table 1
Table 2
The present invention is not limited to above-mentioned embodiment, in the case of without departing substantially from the substantive content of the present invention, this area skill
Any deformation, improvement, the replacement that art personnel are contemplated that each fall within the scope of the present invention.
Claims (10)
1. a kind of preparation method of silkworm chrysalis active peptide, it comprises the following steps：
A, dried silkworm chrysalis meal is added into water, adjusts pH to 8~9, soaked 18~24 hours, separation of solid and liquid, take supernatant；
B, supernatant regulation pH to 4~6 is precipitated, separation of solid and liquid, takes solid portion；
C, the solid portion is added into water, 15~30min is heated at 75~100 DEG C, homogeneous is then carried out, is suspended
Liquid；
D, the suspension is adjusted into pH to 5~10, adds enzyme preparation and digested；The enzyme preparation is by flavor protease, wood
Melon protease and neutral proteinase composition, and the weight ratio of the flavor protease, papain and neutral proteinase be 1~
2: 1~2: 1~2, hydrolysis temperature is 40~65 DEG C, and enzymolysis time is 1.5~8h, obtains enzymolysis liquid；
E, by the enzymolysis liquid destroy the enzyme treatment, silkworm chrysalis active peptide powder is then made.
2. preparation method according to claim 1, it is characterised in that in step B, adjusted the supernatant using weak acid
PH to 4~6；The weak acid is selected from least one of acetic acid, lactic acid, citric acid.
3. preparation method according to claim 1, it is characterised in that in step E, the temperature of the destroy the enzyme treatment for 90~
125 DEG C, the time is 0.5~3h.
4. the preparation method of silkworm chrysalis active peptide according to claim 1, it is characterised in that in step E, at the enzyme deactivation
Enzymolysis liquid concentration after reason, emulsifying, is then spray-dried, obtains silkworm chrysalis active peptide powder；The spray drying is 80~105
Carried out at DEG C.
5. the preparation method of silkworm chrysalis active peptide according to claim 1, it is characterised in that in step E, at the enzyme deactivation
Enzymolysis liquid after reason is centrifuged, and centrifugate filters through plate and frame filter press first, then through ceramic membrane filter, obtains active peptide transmission
Liquid；The active peptide filtrate is concentrated, spray drying, so as to obtain silkworm chrysalis active peptide powder；The spray drying is at 80~105 DEG C
Lower progress；Containing combination filtration adjuvant in the plate and frame filter press, the combination filtration adjuvant is by activated carbon, perlite and diatom
Soil is according to weight than 6~7: 1~2: 2~3 compositions.
6. a kind of silkworm chrysalis active peptide, it is characterised in that the silkworm chrysalis active peptide is used according to any one of Claims 1 to 5
Method be prepared.
A kind of 7. composition of active peptide containing silkworm chrysalis, it is characterised in that the composition by the parts by weight of silkworm chrysalis active peptide 90~98,
The parts by weight of mulberries 1~5 and the parts by weight of spina date seed 1~5 are made.
8. the preparation method of the composition of the active peptide according to claim 7 containing silkworm chrysalis, it is characterised in that the preparation side
Method includes：(1) method according to any one of Claims 1 to 5 prepares silkworm chrysalis active peptide powder；(2) respectively by mulberries and acid
Jujube kernel is prepared as mulberries powder and spiny jujuba seed powder；(3) silkworm chrysalis active peptide powder, mulberries powder and the spiny jujuba seed powder are well mixed, obtained
To the composition；Or
The preparation method includes：(1) method according to any one of Claims 1 to 5 prepares silkworm chrysalis active peptide powder；(2)
By the use of water as solvent extraction after mulberries and spina date seed are mixed, extract solution concentration, it dry, pulverize, obtain medicinal substances extract；(3)
The silkworm chrysalis active peptide powder and medicinal substances extract are well mixed, obtain the composition.
9. the composition of a kind of active peptide containing silkworm chrysalis, it is characterised in that the composition is by the parts by weight of dried silkworm chrysalis meal 90~98, mulberries
1~5 parts by weight and the parts by weight of spina date seed 1~5 are made.
10. the preparation method of the composition of the active peptide according to claim 9 containing silkworm chrysalis, it is characterised in that the preparation
Method comprises the following steps：
A ', dried silkworm chrysalis meal is added to water, adjust pH to 8~9, soaked 18~24 hours, separation of solid and liquid, take supernatant；
B ', by the supernatant regulation pH to 4~6 precipitated, separation of solid and liquid, take solid portion；
C ', by the solid portion add water, at 75~100 DEG C heat 15~30min, then carry out homogeneous, be suspended
Liquid；
D ', the suspension is adjusted to pH to 5~10, addition enzyme preparation is digested；The enzyme preparation is by flavor protease, wood
Melon protease and neutral proteinase composition, and the weight ratio of the flavor protease, papain and neutral proteinase be 1~
2: 1~2: 1~2；Hydrolysis temperature is 40-65 DEG C, enzymolysis time 1.5-8h, obtains enzymolysis liquid；
D1 ', the mulberries and spina date seed added in the enzymolysis liquid, heating and refluxing extraction, filtering, obtain mixed liquor；
E ', the composition that the mixed liquor is made to the active peptide containing silkworm chrysalis.
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| CN110477381A (en) * | 2019-09-03 | 2019-11-22 | 广东紫薇星健康产业有限公司 | A kind of albumin amino acid Calcium oral liquid prescription and preparation method with immunological regulation healthcare function |
| CN111387338A (en) * | 2020-04-08 | 2020-07-10 | 上海辰钿实业有限公司 | Preparation method of pepper seed active protein peptide |
| CN112335888A (en) * | 2020-11-09 | 2021-02-09 | 四川必信科技有限公司 | Sea cucumber and abalone oligopeptide powder and preparation method thereof |
| CN113897687A (en) * | 2021-10-20 | 2022-01-07 | 浙江理工大学 | A kind of high value-added raw silk processing method |
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| CN1896267A (en) * | 2006-06-23 | 2007-01-17 | 浙江大学 | Preparation of depressor peptide by silkworm chrysalis |
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| CN101748180A (en) * | 2010-01-18 | 2010-06-23 | 山东世纪春食品有限公司 | Functional peanut small molecule mixed polypeptide, preparation method and application thereof |
| CN104789629A (en) * | 2015-04-30 | 2015-07-22 | 中国食品发酵工业研究院 | Chickpea oligopeptide and preparation method thereof |
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| CN110477381A (en) * | 2019-09-03 | 2019-11-22 | 广东紫薇星健康产业有限公司 | A kind of albumin amino acid Calcium oral liquid prescription and preparation method with immunological regulation healthcare function |
| CN111387338A (en) * | 2020-04-08 | 2020-07-10 | 上海辰钿实业有限公司 | Preparation method of pepper seed active protein peptide |
| CN112335888A (en) * | 2020-11-09 | 2021-02-09 | 四川必信科技有限公司 | Sea cucumber and abalone oligopeptide powder and preparation method thereof |
| CN113897687A (en) * | 2021-10-20 | 2022-01-07 | 浙江理工大学 | A kind of high value-added raw silk processing method |
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