CN107974479A - A kind of method that bird's nest oligopeptide is prepared with microwave and membrane technology - Google Patents
A kind of method that bird's nest oligopeptide is prepared with microwave and membrane technology Download PDFInfo
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- CN107974479A CN107974479A CN201711215354.8A CN201711215354A CN107974479A CN 107974479 A CN107974479 A CN 107974479A CN 201711215354 A CN201711215354 A CN 201711215354A CN 107974479 A CN107974479 A CN 107974479A
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
- C12P21/06—Preparation of peptides or proteins produced by the hydrolysis of a peptide bond, e.g. hydrolysate products
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
Abstract
The invention belongs to bird's nest product deep process technology field, more particularly to a kind of method that bird's nest oligopeptide is prepared with microwave and membrane technology.A kind of method that bird's nest oligopeptide is prepared with microwave and membrane technology, includes the following steps：Bird's nest is first crushed to 80 mesh, soaks swollen hair with hot deionized water, is starched with colloid mill circular grinding into by the bird's nest of 120 mesh, addition deionized water makes the solid-liquid of slurries reach 1：30, with the pH value of dilute NaOH solution adjusting slurry to 10.6, flow out protein with microwave treatment, in microwave radiation technology double-enzyme hydrolysis, enzymolysis liquid is through microwave, 2 grades of ultrafiltration and nanofiltration aggregation bird's nest oligopeptide, oligopeptide liquid is concentrated, freezed again, low-temperature microwave drying, obtains the oligomeric peptide product of bird's nest.Peptide product prepared by the method for the present invention, its peptide fragment are distributed in 5 peptide, 16 peptide, 539 1745Da of relative molecular weight of peptide.
Description
Technical field
The invention belongs to bird's nest product deep process technology field, more particularly to microwave and membrane technology to prepare bird's nest low for one kind
The method of poly- peptide.
Background technology
Bird's nest is eaten as noble name, and the either salty food of method or sweet food of the edible bird's nest of tradition, it is all first that it, which is pre-processed,
Swollen hair, swollen hair is soaked, alkali is sent out and evaporation；In addition to sending out or evaporate using alkali when urgent need, usually all use and be soaked.Be soaked is first to use
Cold clear water or the immersion of warm clear water, after it is impregnated with swollen hair, selects net fine hair, impurity and rotten part, then match somebody with somebody by the requirement of dietotherapy
With various seasonings, cooked slowly with long period small fire, in nutrition and physiology also underdeveloped epoch, so culinary art has been calculated
It is upper advanced reasonable.But in terms of the scientific and technological level angle of today, there are many deficiencies, or even say that it is a little to waste preciousness that must weigh
Component, as the protein degradation in vivo in bird's nest will consume certain energy, and cannot degradable macro-molecular protein,
Bioactive substance in bird's nest fails to dissociate to be absorbed, its physiological function cannot play a role or improperly cook
In lose or reduce its activity, and biological value cannot be produced.
Modern nutriology research shows that protein is one of most important organic substance in mechanism eucaryotic cell structure, in food
Protein its molecular weight minimum have ten hundreds of dalton, generally all in hundreds of thousands even more than million.These albumen
After matter enters in vivo, enzymatically hydrolyze and (promote the collective effects such as enzyme, pepsin, pancreatin, hydrochloric acid in gastric juice, digestive ferment), protein is dropped
Solve as free amino acid, amino acid synthetic peptide again, eventually through intestinal absorption, rather than has previously been thought that intestinal absorption free amine group
Acid.
Peptide is also stranger at present for ruck.The research of peptide science, is obtained in history in the last hundred years for nearly 50 years
The scientist for obtaining Nobel Prize in Physiology of Medicine has 24.Scientist's Chinese descendant in America Nobel laureate's Steven Chu once broke
Speech, 21 century is the century of peptide.Numerous studies confirm：Generation, development, treatment and the rehabilitation of all diseases are all related with peptide.It is several
All cells are all adjusted by peptide, it is related to the every field such as hormone, nerve, cell growth and reproduction.Peptide is by 2 or 2
The material that identical or different amino acid residue is connected in a manner of peptide bond more than a, its molecular structure is between protein
A kind of compound between amino acid, is the fragment of protein, since the grade of composition amino acid sequence and structure is different,
Make this material that there is the physiological function in terms of necessarily.Only with regard to body absorption aspects, we look at its absorption characteristics：1) it is not required to disappear
Change system is digested, and is directly absorbed；2) quick, water consumption energy is absorbed；3) absorbed in the form of complete；4) it is very complete
Absorb；5) after absorbing waste is not produced without metabolism；6) actively absorbed；7) preferential absorption；8) carrier and translocation, energy are played
By various nutritional ingredients load, be adhered on body, bring into each organ, tissue in it is intracellular, absorbed together with body；
9) courier's mediator is played the role of in vivo after absorbing.Above-mentioned mechanism of absorption, declines, the people of obstacle for digestive system function
Group or patient, as nutrition are optimal Opsonizing methods using peptide matters.Therefore, if bird's nest protein can be changed into class chemical combination
Thing will increase substantially the biological value of bird's nest.
At present, the method that bird's nest peptide is prepared with bird's nest, there are no disclosed document, and the product of benevolence hall sale is only supported in Indonesia
《The prebiotic bird's nest collagen peptide of swallow peptide》Claim to be to obtain small molecule bird's nest powder with modern low-temperature extraction technology on specification, and largely
Retain the components such as epidermal growth factor.
The information for being disclosed in the background section is merely intended to understanding of the increase to the general background of the present invention, without answering
It has been the prior art well known to persons skilled in the art when being considered as recognizing or implying the information structure in any form.
The content of the invention
For overcome the deficiencies in the prior art, the present invention provides a kind of side that bird's nest oligopeptide is prepared with microwave and membrane technology
Method.
Technical solution provided by the invention is as follows：
A kind of method that bird's nest oligopeptide is prepared with microwave and membrane technology, includes the following steps：
(1) first bird's nest is crushed with pulverizer, obtains bird's nest powder；
(2) bird's nest powder is added in deionized water, soaks 3-4h, bird's nest powder is expanded to 5-6 times, into translucent glue
Plastochondria；
(3) soaked bird's nest powder is circulated into barreling 8-10min together with soaking water with colloid mill, obtains bird's nest slurry；
(4) deionized water is added in ground bird's nest slurry, solid-to-liquid ratio is reached 1：30, then sized mixing with dilute NaOH solution
Liquid pH value mixes again to 10.0 in colloid；
(5) alkaline bird's nest slurries are placed in the microwave that microwave frequency is 2450mHz to occur in jar, it is total starts microwave treatment
Time 30min, 60 DEG C of temperature control, keeps the temperature 30min at 50-60 DEG C of temperature, detects the pH value of slurries, work as pH value<When 8.4, wait to starch
Liquid cools down<45 DEG C, add alkali protease, while use dilute NaOH tune pH value to 9.0, startup microwave treatment, total microwave time 25-
30min, 60 DEG C of temperature control, keeps the temperature 20min between 50-60 DEG C；
(6) above-mentioned steps (5) are treated that the temperature of feed liquid is down to after the hydrolysis liquid insulation of alkaline enzyme microwave radiation technology<At 45 DEG C,
Neutral proteinase is added, starts microwave treatment, microwave total time 20min, 55 DEG C of temperature control, keeps the temperature 20min between 50-55 DEG C, inspection
Material liquid pH value is surveyed, works as pH value<When 7.2, terminate enzymolysis process；
(7) enzymolysis liquid is discharged, be cooled to<After 50 DEG C, with the pH value of acetic acid tune enzymolysis liquid to 6.0-6.5, with hollow fibre
Tie up film and carry out micro-filtration, dope collection is stand-by, and filtrate does lower 2 grades of ultrafiltration of step；
(8) ultrafiltration is carried out with the ultrafiltration membrane that can block molecular weight 10000Da and 2000Da；10000Da Filtration Controls are pumped
0.2-0.4MPa, 40-45 DEG C of temperature, film surface flow 1-1.5m/s, 3-4 times of cocnentration factor；2000Da Filtration Controls are pumped 0.3-
0.5MPa, 40-45 DEG C of temperature, film surface flow 1m/s, 4-5 times of cocnentration factor；
(9) permeate of 2000Da ultrafiltration, with the nanofiltration membrane of 500Da, control pump pressure 0.5-0.8MPa, circular flow
1-2.5m3/ h, 5-7 times of cocnentration factor；The dope of nanofiltration works as concentration<Decompression is sent to when Be20 is spent, is further concentrated to temperature control when Be25 is spent
58 DEG C, discharge is sent to after being cooled to room temperature in the refrigerator-freezer or freezer of energy depth, is refrigerated to -35 DEG C, and keep 1-2h；When nanofiltration
Dense concentrate concentration>When Be20 is spent, freezing is directly sent to without be concentrated under reduced pressure；
(10) the dope ice cube freezed, taking-up are broken into fritter, are placed in swinging microwave vacuum dryer and are dehydrated, and do
It is dry to moisture content be less than 6% after, terminate drying process, obtain loose shape crystalline product, be bird's nest oligopeptide.
Preferably, the fineness of bird's nest powder is 80 mesh in step (1).
Preferably, the temperature of deionized water is 50-60 DEG C in step (2).
Preferably, the solid-to-liquid ratio of bird's nest powder and deionized water is 1 in step (2):5-6.
Preferably, the fineness that bird's nest is starched in step (3) is 120 mesh.
Preferably, the additive amount of step (5) neutral and alkali protease is the 3.5% of substrate weight, the alkali protease
Enzyme activity be 105U/g。
Preferably, the additive amount of the neutral proteinase in step (6) is the 2.5% of substrate weight, the neutral protein
The enzyme activity of enzyme is 4000U/g.
Preferably, the vacuum of swinging microwave vacuum dryer is 0.05-0.08MPa in step (10), temperature is
58℃。
Compared with prior art, the present invention has the advantages that：
(1) present invention is studied by a large number of experiments, and the oligomeric Gly-His-Lys of bird's nest are obtained with enzyme process, whole low temperature.
(2) method of the invention prepares gained peptide product, its peptide fragment is distributed in -16 peptide of 5 peptide, and the relative molecular weight of peptide exists
Between 539-1745Da.
Brief description of the drawings
Fig. 1 is the process flow chart that the present invention prepares bird's nest oligopeptide with microwave and membrane technology.
Embodiment
Following embodiments do not limit the present invention to be better understood from the present invention.Experimental method in following embodiments,
Unless otherwise specified, it is conventional method.
Embodiment 1：
A kind of method that bird's nest oligopeptide is prepared with microwave and membrane technology, includes the following steps：
(1) bird's nest is crushed to 80 mesh into bird's nest powder；
(2) solid-liquid 1 is pressed with 50-60 DEG C of deionization hot water：5-6 soaks 3-4h, bird's nest powder is expanded to 5-6 times into semi-transparent
Bright shape micelle body；
(3) soaked bird's nest powder is circulated into barreling 8-10min together with soaking water with colloid mill, made into by 120 purposes
Bird's nest is starched；
(4) ground bird's nest is starched, addition deionized water makes solid-to-liquid ratio reach 1：30, with dilute NaOH adjusting slurries pH value extremely
10.0, mixed again in colloid；
(5) by alkaline bird's nest slurries, it is placed in the microwave that microwave frequency is 2450mHz and occurs in jar, start microwave treatment
Total time 30min, 60 DEG C of temperature control, 30Min is kept the temperature between 50-60 DEG C, the pH value of slurries is detected, works as pH value<When 8.4, slurries are treated
Cooling<45 DEG C, add the alkali protease (enzyme activity 10 by substrate weight 3.5%5U/g), while with dilute NaOH tune pH value extremely
9.0, start microwave treatment, total microwave time 25-30min, 60 DEG C of temperature control, keeps the temperature 20min between 50-60 DEG C；
(6) after alkaline enzyme microwave radiation technology hydrolysis liquid insulation being obtained through above-mentioned steps (5), treat that the temperature of feed liquid is down to<45
DEG C when, addition neutral enzymatic (enzyme activity 4000U/g) press amount of substrate 2.5%, start microwave treatment, microwave total time 20Min, control
55 DEG C of temperature, keeps the temperature 20min between 50-55 DEG C, examines material liquid pH value, works as pH value<When 7.2, terminate enzymolysis process；
(7) enzymolysis liquid is discharged, be cooled to<After 50 DEG C, with the pH value of acetic acid tune enzymolysis liquid to 6.0-6.5, with hollow fibre
Tie up film and carry out micro-filtration, dope collection is stand-by, and filtrate does lower 2 grades of ultrafiltration of step；
(8) ultrafiltration is carried out with the ultrafiltration membrane that can block molecular weight 10000Da and 2000Da.10000Da Filtration Controls are pumped
0.2-0.4MPa, 40-45 DEG C of temperature, film surface flow 1-1.5m/s, 3-4 times of cocnentration factor；2000Da Filtration Controls are pumped 0.3-
0.5MPa, 40-45 DEG C of temperature, film surface flow 1m/s, 4-5 times of cocnentration factor；
(9) permeate of 2000Da ultrafiltration, with the nanofiltration membrane of 500Da, control pump pressure 0.5-0.8MPa, circular flow
1-2.5m3/ h, 5-7 times of cocnentration factor；The dope of nanofiltration works as concentration<Decompression is sent to when Be20 is spent, is further concentrated to Be25 degree temperature control 58
DEG C, discharge is sent to after being cooled to room temperature to be refrigerated to -35 DEG C, and keep 1-2h in the refrigerator-freezer or freezer of depth, dense when nanofiltration
Concentrate concentration>When Be20 is spent, freezing can be directly sent to without being concentrated under reduced pressure；
(10) the dope ice cube freezed, taking-up are broken into fritter, are placed in swinging microwave vacuum dryer and are dehydrated, and do
It is dry to moisture content be less than 6% after, terminate drying process, obtain loose shape crystalline product, be bird's nest oligopeptide；
(11) micro-filtration dope, 2 grades of ultrafiltration dopes, nanofiltration permeate liquid are merged, is performed under reduced pressure concentration, temperature control 60
DEG C, concentrate is obtained, is freezed by step (9) and (10), drying means operation, obtains crystalloid bird's nest powder.
Technique as stated above is operated, and Sialic acid and bird's nest oligopeptide, bird's nest can be produced by making tune in nanofiltration process
Sour product mix.
1) when Sialic acid to be extracted, increase level-one nanofiltration in nanofiltration process, isolate 200Da to 500Da molecular weight
Region is bird's nest acid solution.
Specific practice is：2000Da ultrafiltration permeate intercepts material of the molecular weight more than 500Da with 500Da NF membranes, thoroughly
To cross liquid 200Da NF membranes and intercept the material that molecular weight is more than 200Da, the dope of 200Da nanofiltrations is Sialic acid pregnant solution, because
Relative molecular weight for Sialic acid is 309.2699Da.By the concentration of Sialic acid pregnant solution, freeze, dry, up to bird's nest acid product.
2) when bird's nest oligopeptide to be produced, Sialic acid product mix into, the NF membrane of 500Da is changed to the nanofiltration of 200Da
Film, the ultrafiltration permeate of 2000Da are retained by 200Da NF membranes, obtain the material of dope, its molecular weight in 200Da extremely
Between 2000Da, the component of the Sialic acid of bird's nest oligopeptide, concentrated, freezing, drying, up to bird's nest oligopeptide and swallow are enriched
The product mix of nest acid.
With the technique and operating condition in above-mentioned the method for the present invention, bird's nest raw material 2000g is thrown, available bird's nest peptide, its
Peptide fragment is distributed and occupancy volume is shown in Table 1.
The peptide fragment distribution for the bird's nest peptide that the method for the present invention of table 1 obtains and occupancy volume measure
| Peptide fragment | Sequence number (a) | Account for total peptide amount (%) |
| 5 peptides | 18 | 0.79 |
| 6 peptides | 79 | 17.67 |
| 7 peptides | 85 | 25.38 |
| 8 peptides | 77 | 17.67 |
| 9 peptides | 51 | 12.84 |
| 10 peptides | 30 | 6.23 |
| 11 peptides | 17 | 2.07 |
| 12 peptides | 9 | 1.01 |
| 13 peptides | 11 | 1.93 |
| 14 peptides | 9 | 15.35 |
| 15 peptides | 4 | 6.07 |
| 16 peptides | 2 | 6.31 |
As shown in Table 1, peptide fragment (is less than 5 peptides and does not have more than 16 peptides) from 5 peptides to 16 peptides, shares 392 amino acid sequences
Row, the total peptide amount 72.50% of 6 peptides to 10 peptides, 5 peptides to 12 peptides account for total peptide amount 75.58%, and bird's nest enzymolysis gained is seen from the distribution of peptide amount
Peptide, which is seen, substantially belongs to oligopeptide scope.
It is foregoing to the present invention specific exemplary embodiment description be in order to illustrate and illustration purpose.These descriptions
It is not wishing to limit the invention to disclosed precise forms, and it will be apparent that according to the above instruction, can be much changed
And change.The purpose of selecting and describing the exemplary embodiment is that explain that the certain principles of the present invention and its reality should
With so that those skilled in the art can realize and utilize the present invention a variety of exemplaries and
Various chooses and changes.The scope of the present invention is intended to be limited by claims and its equivalents.
Claims (8)
- A kind of 1. method that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that include the following steps：(1) first bird's nest is crushed with pulverizer, obtains bird's nest powder；(2) bird's nest powder is added in deionized water, soaks 3-4h, bird's nest powder is expanded to 5-6 times, into translucent micelle Body；(3) soaked bird's nest powder is circulated into barreling 8-10min together with soaking water with colloid mill, obtains bird's nest slurry；(4) deionized water is added in ground bird's nest slurry, solid-to-liquid ratio is reached 1：30, then with dilute NaOH solution adjusting slurry pH Value mixes again to 10.0 in colloid；(5) alkaline bird's nest slurries are placed in the microwave that microwave frequency is 2450mHz to occur in jar, start microwave treatment total time 30min, 60 DEG C of temperature control, keeps the temperature 30min at 50-60 DEG C of temperature, detects the pH value of slurries, work as pH value<When 8.4, treat that slurries are cold But<45 DEG C, add alkali protease, while use dilute NaOH tune pH value to 9.0, startup microwave treatment, total microwave time 25- 30min, 60 DEG C of temperature control, keeps the temperature 20min between 50-60 DEG C；(6) above-mentioned steps (5) are treated that the temperature of feed liquid is down to after the hydrolysis liquid insulation of alkaline enzyme microwave radiation technology<At 45 DEG C, addition Neutral proteinase, starts microwave treatment, microwave total time 20min, 55 DEG C of temperature control, keeps the temperature 20min, detection material between 50-55 DEG C Liquid pH value, works as pH value<When 7.2, terminate enzymolysis process；(7) enzymolysis liquid is discharged, be cooled to<After 50 DEG C, with the pH value of acetic acid tune enzymolysis liquid to 6.0-6.5, hollow-fibre membrane is used Micro-filtration is carried out, dope collection is stand-by, and filtrate does lower 2 grades of ultrafiltration of step；(8) ultrafiltration is carried out with the ultrafiltration membrane that can block molecular weight 10000Da and 2000Da；10000Da Filtration Controls are pumped 0.2- 0.4MPa, 40-45 DEG C of temperature, film surface flow 1-1.5m/s, 3-4 times of cocnentration factor；2000Da Filtration Controls are pumped 0.3-0.5MPa, 40-45 DEG C of temperature, film surface flow 1m/s, 4-5 times of cocnentration factor；(9) permeate of 2000Da ultrafiltration, with the nanofiltration membrane of 500Da, control pump pressure 0.5-0.8MPa, circular flow 1- 2.5m3/ h, 5-7 times of cocnentration factor；The dope of nanofiltration works as concentration<Decompression is sent to when Be20 is spent, is further concentrated to temperature control 58 when Be25 is spent DEG C, discharge is sent to after being cooled to room temperature in the refrigerator-freezer or freezer of energy depth, is refrigerated to -35 DEG C, and keep 1-2h；It is dense when nanofiltration Concentrate concentration>When Be20 is spent, freezing is directly sent to without be concentrated under reduced pressure；(10) the dope ice cube freezed, taking-up are broken into fritter, are placed in swinging microwave vacuum dryer and are dehydrated, and drying is extremely After moisture content is less than 6%, terminates drying process, obtain loose shape crystalline product, be bird's nest oligopeptide.
- 2. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (1) fineness of bird's nest powder is 80 mesh in.
- 3. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (2) temperature of deionized water is 50-60 DEG C in.
- 4. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (2) solid-to-liquid ratio of bird's nest powder and deionized water is 1 in:5-6.
- 5. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (3) fineness that bird's nest is starched in is 120 mesh.
- 6. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (5) additive amount of neutral and alkali protease is the 3.5% of substrate weight, and the enzyme activity of the alkali protease is 105U/g。
- 7. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (6) additive amount of the neutral proteinase in is the 2.5% of substrate weight, and the enzyme activity of the neutral proteinase is 4000U/g.
- 8. the method according to claim 1 that bird's nest oligopeptide is prepared with microwave and membrane technology, it is characterised in that step (10) vacuum of swinging microwave vacuum dryer is 0.05-0.08MPa in, and temperature is 58 DEG C.
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| CN109464359A (en) * | 2018-11-21 | 2019-03-15 | 佛山科学技术学院 | Skin care item and preparation method thereof containing bird's nest peptide |
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| CN110257459A (en) * | 2019-05-16 | 2019-09-20 | 广西慧宝源医药科技有限公司 | Bird's nest small-molecular peptides and its instant Gly-His-Lys |
| CN111662945A (en) * | 2020-07-13 | 2020-09-15 | 广西赣华生物科技有限公司 | Method for extracting noni peptide by microwave-assisted enzymolysis method and noni peptide powder |
| CN111748013A (en) * | 2020-07-13 | 2020-10-09 | 广西赣华生物科技有限公司 | Natural noni peptide extraction method and natural noni peptide powder |
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| CN110257459A (en) * | 2019-05-16 | 2019-09-20 | 广西慧宝源医药科技有限公司 | Bird's nest small-molecular peptides and its instant Gly-His-Lys |
| CN110257459B (en) * | 2019-05-16 | 2021-04-13 | 广西慧宝源医药科技有限公司 | Bird's nest small molecular peptide and instant peptide powder thereof |
| CN111662945A (en) * | 2020-07-13 | 2020-09-15 | 广西赣华生物科技有限公司 | Method for extracting noni peptide by microwave-assisted enzymolysis method and noni peptide powder |
| CN111748013A (en) * | 2020-07-13 | 2020-10-09 | 广西赣华生物科技有限公司 | Natural noni peptide extraction method and natural noni peptide powder |
| CN111662945B (en) * | 2020-07-13 | 2023-08-11 | 广西赣华生物科技有限公司 | Method for extracting noni peptide by microwave-assisted enzymolysis and noni peptide powder |
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