CN107556364A - Subcritical water assistance enzymolysis extracts the method and product of abalone proteins peptide - Google Patents
Subcritical water assistance enzymolysis extracts the method and product of abalone proteins peptide Download PDFInfo
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- CN107556364A CN107556364A CN201710916485.2A CN201710916485A CN107556364A CN 107556364 A CN107556364 A CN 107556364A CN 201710916485 A CN201710916485 A CN 201710916485A CN 107556364 A CN107556364 A CN 107556364A
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Abstract
The invention discloses the method and product of a kind of subcritical water assistance enzymolysis extraction abalone proteins peptide, technical matters is included using abalone internal organ as raw material, extracting protein first is handled to it using subcritical water, then the albumen extracted recycles pectase and protease to be digested, concentrated by column chromatography for separation, NF membrane, vacuum concentration and spray drying prepare abalone proteins peptide powder, and utilize abalone proteins peptide powder development function health product.The technology of the present invention improves the technology that tradition enzymolysis prepares protein peptides, extracts abalone proteins peptide using subcritical water assistance enzymolysis, can not only reduce the usage amount of enzyme, the production time can also be greatly shortened, reduces production cost.Moreover, the abalone proteins peptide prepared has good anti-oxidant, anti-tumor function, the application for abalone proteins peptide provides condition.
Description
Technical field
The present invention relates to a kind of technology of subcritical water assistance enzymolysis extraction abalone proteins peptide and products thereof exploitation, specifically
The method and product for being related to a kind of subcritical water assistance enzymolysis extraction abalone proteins peptide are said, it is used at HTHP subcritical water
Reason is prepared by enzymatic isolation method again from abalone internal organ extracting protein has biologically active protein, peptide, and utilizes the abalone proteins peptide of extraction
The technology of development function health product.
Background technology
Subcritical water is also referred to as pressure hot water, and its temperature keeps water between 100~374 DEG C, by control system pressure
For liquid condition.The dielectric constant of subcritical water is low, rich in hydrogen ion and hydroxide ion, therefore has acid-base catalysis function,
Can be with fast decoupled organic matter.Compared with traditional extracting method, the method extracted using subcritical water have cost it is low,
The advantages that energy consumption is low, active component recovery rate is high, extraction rate is fast and economic and environment-friendly.On the other hand, processed in abalone
Cheng Zhong, substantial amounts of abalone internal organ will necessarily be produced, account for abalone weight 25%.Abalone internal organ are rich in albumen and polysaccharide, but main at present
It is used to produce the low value-added products such as flavoring, feed.Although reporting both at home and abroad abalone is extracted using mode of action
Internal organ active material, applicant also disclose the coproduction preparation side of abalone polysaccharide, lipid and protein peptides in early stage using mode of action
Method（The A of China Patent Publication No. CN 103255186）, but due to the reason such as enzymolysis time is long, recovery rate is low, in actual applications
The problems such as production cost is high also be present.
Therefore, the present invention is first extracted using subcritical water to abalone internal organ, and the albumen of extraction is entered by enzymatic isolation method
Row enzymolysis, then abalone proteins peptide is refined using activated carbon decolorizing and column chromatography technology, by being spray-dried the Bao prepared
Fish protein peptide has good anti-oxidant and anti-cancer function, and utilizes the abalone proteins peptide development function health product prepared.
The content of the invention
It is an object of the present invention to provide a kind of method of subcritical water assistance enzymolysis extraction abalone proteins peptide, it is
Using abalone internal organ as raw material, mainly being prepared using HTHP subcritical water aid in treatment and enzymatic isolation method extracting protein has biology
The abalone proteins peptide of activity, recycle the technology of the abalone proteins peptide development function health product of extraction.
It is another object of the present invention to all preferable subcritical water assistance enzymolysis extraction of a kind of functional activity and mouthfeel
Product made of abalone proteins peptide.
To achieve these goals, the technical scheme is that：
The present invention is a kind of method of subcritical water assistance enzymolysis extraction abalone proteins peptide, is comprised the following steps：(1) it is subcritical
Water process：Abalone internal organ add pure water by 13 ~ 15 (w/v) solid-liquid ratios, and 15 min ~ 60 are extracted under subcritical water condition
Min, it is cooled to after room temperature and obtains protein extracting liquid with stainless steel filter screen removing filter residue, filter residue is by that can be used as sea after drying
Algae fertilizer；(2) digest：Complex enzyme is added in the protein extracting liquid that step (1) obtains, complex enzyme addition is protein extracting liquid
0.1% ~ 0.5% (w/v), the h of the h of enzyme digestion reaction 1 ~ 3；(3) separate, concentrate and be spray-dried：The albumen that step (2) is obtained
After enzymolysis liquid is decolourized using activated carbon and resin, deodorant, removing heavy metal, dearsenification etc. are refined, concentrated by NF membrane, very
Sky concentration and spray drying are prepared into abalone proteins peptide powder；(4) production：Add in the abalone proteins peptide powder of extraction
Add beta-schardinger dextrin, collagen peptide, soybean lecithin, blue or green olive pulp extract and honey, be well mixed, product particle is made, then
1% ~ 5% magnesium stearate of product particle quality is added, is well mixed, is pressed into chewable tablets.
In step (1), described subcritical water refers to that temperature is 120 DEG C ~ 160 DEG C, and pressure is 1.0 MPa ~ 7.0
MPa water.
In step (2), described complex enzyme refers to papain, flavor protease, neutral proteinase and pineapple
Any of which such as protease protease is compounded with pectase, and the compound proportion of protease and pectase is 3:1~5:1；.
In step (2), the enzymatic hydrolysis condition of described complex enzyme is pH 5.0 ~ 7.0,40 DEG C ~ 50 DEG C of temperature.
In step (3), described activated carbon decolorizing deodorant condition is：The pH of protein enzymatic hydrolyzate is adjusted to 5.0 ~ 6.0, activity
The addition of charcoal is 0.5% ~ 2.0% (w/v) of protein enzymatic hydrolyzate volume, the min of 15 min ~ 60 is incubated at 40 DEG C ~ 50 DEG C, so
Afterwards activated carbon is removed using disk centrifugal separator；Described resin deodorant removes heavy metal dearsenification condition：By macroporous absorbent resin,
The mixed fillers of the chelating resin of heavy metal and the chelate fibre composition of arsenic-adsorbing are adsorbed, its mixed proportion is 7:2:1；Protease
The pH for solving liquid is 5.0 ~ 6.0, and protein enzymatic hydrolyzate residence time in mixed fillers is the min of 15 min ~ 30.
In step (4), the constituent content of described product particle by weight is：Abalone proteins peptide powder 20% ~ 30%,
Beta-schardinger dextrin 55% ~ 70%, collagen peptide 2% ~ 10%, soybean lecithin 3% ~ 5%, blue or green olive pulp extract 1.0% ~ 2.5%, honey
0.5%~2.0%。
Described abalone internal organ are one in fresh abalone internal organ, frozen abalone internal organ or heat-treated abalone internal organ
Kind.
The present invention is product made of a kind of subcritical water assistance enzymolysis extraction abalone proteins peptide, its component by weight
Content is：Abalone proteins peptide powder 20% ~ 30%, beta-schardinger dextrin 55% ~ 70%, collagen peptide 2% ~ 10%, soybean lecithin 3% ~ 5%, green grass or young crops
Olive pulp extract 1.0% ~ 2.5%, honey 0.5% ~ 2.0%.
After such scheme, the albumen in Subcritical Water Extraction abalone internal organ is first used due to the present invention, then pass through enzyme
Solution method prepares protein peptides, it is possible to reduce the dosage and enzymolysis time of enzyme, and the temperature of subcritical water that uses of the present invention and
Pressure, which will not make not digest complete marine alga in abalone internal organ, to be dissolved, and the separation of follow-up enzymolysis liquid avoids the need for passing through ceramics
Film and milipore filter are separated, and production time and the production cost of abalone proteins peptide can be greatly shortened.Moreover, abalone internal organ
In marine alga a certain degree of digestion is all passed through by abalone, the filter residue after protein extracting can be directly as alga fertilizer by drying
Use, reduce the discharge of discarded object.Simultaneously as after protein enzymatic hydrolyzate is by activated carbon decolorizing deodorant in the present invention, then adopt
The decoloration deodorization, removing heavy metal and dearsenification of profound level are carried out to it with hybrid resin, can not only improve the quality of product, also
It can guarantee that the security of product.On the other hand, with the deterioration of environment, the quickening of rhythm of life and the raising of health care consciousness,
Concern of the people to health gradually extends to sub-health state from disease, therefore the product with healthcare function is increasingly subject to consume
The concern of person.For the present invention on the basis of the abalone proteins peptide containing the bioactivity such as anti-oxidant and antitumor, addition can be by human body
The small molecule collagen oligopeptide and plant essence soybean lecithin absorbed, and there is improving effect to abalone proteins peptide flavor
Blue or green olive pulp extract and honey, and be first prepared into little particle with beta-schardinger dextrin, then tabletting is carried out with magnesium stearate, make Bao
The functional activity and mouthfeel of fish protein peptide product can be further enhanced, and be easily esthetically acceptable to the consumers, and be had good
Market prospects.Therefore the present invention has advantages below：
1st, the present invention can be used for producing alga fertilizer using abalone internal organ extraction abalone proteins peptide, residue, can reach efficiently comprehensive
Close and utilize, reduce environmental pollution.
2nd, the present invention can not only reduce the usage amount of enzyme using subcritical water assistance enzymolysis extraction abalone proteins peptide, may be used also
So that the production time is greatly shortened, and then reduce production cost.
3rd, abalone proteins peptide prepared by the present invention has good anti-oxidant, anti-tumor function.
4th, the product function activity that the present invention is prepared with abalone proteins peptide is strengthened, it is not easy to the moisture absorption, it is in good taste, and in reality
Easy popularization and application in border.
The present invention is further illustrated for table and specific embodiment below in conjunction with the accompanying drawings.
Accompanying drawing table explanation
Fig. 1 is the antioxidation activity in vitro figure of abalone proteins peptide produced by the invention.
Fig. 2 is influence figure of the abalone proteins peptide produced by the invention to breast cancer MDA-MB-231 cell survival rates.
Table 1 is the basis of abalone proteins peptide produced by the invention.
Table 2 is the molecular weight distribution of the basis of abalone proteins peptide produced by the invention.
Embodiment
First, method：
Embodiment 1
The present invention is a kind of method of subcritical water assistance enzymolysis extraction abalone proteins peptide, is comprised the following steps：
(1) subcritical water is handled：0.4 kg abalones internal organ and 1.2 L pure water, the temperature of reactor are added in pyroreaction kettle
Degree and pressure are improved to 120 DEG C and 7.0 MPa rapidly, and room temperature simultaneously pressure release is cooled to after the min of heat-insulation pressure keeping 60, uses filtered through gauze
Obtain protein extracting liquid and filter residue；
(2) digest：Step (1) is obtained 1.0g papains and 0.2g pectases, enzyme are added in about 1.2L protein extracting liquid
1 h of solution reaction；Complex enzyme zymohydrolysis condition is pH 5.0, temperature 50 C.
(3) separate, concentrate and be spray-dried：6g activated carbons are added in the protein enzymatic hydrolyzate that step (2) obtains, at 50 DEG C
Lower insulation 60min, then remove activated carbon using disk centrifugal separator；Normal temperature is cooled to, pH is confirmed and adjusts to 5.0, by big
Mixed fillers (the mixed proportion 7 of the chelate fibre composition of macroporous adsorbent resin, the chelating resin for adsorbing heavy metal and arsenic-adsorbing:
2:1) column chromatography post decolourized, deodorant, removing heavy metal, dearsenification, residence time of the liquid in chromatographic column be 15 min, sharp
It is 10% to carry out desalination and concentration to solid concentration with the NF membrane that molecular cut off is 150 Da, by being concentrated in vacuo to solid
Thing concentration is 30%, and finally spray drying is prepared into abalone proteins peptide powder；
(4) production：According to abalone proteins peptide powder 20%, beta-schardinger dextrin 70%, glue in the abalone proteins peptide powder of extraction
Former peptide 5%, soybean lecithin 3%, blue or green olive pulp extract 1.0%, the addition of honey 1.0% are well mixed, and particle is made, adds
1% magnesium stearate of granular mass, it is well mixed, is pressed into chewable tablets.
Embodiment 2
The present invention is a kind of method of subcritical water assistance enzymolysis extraction abalone proteins peptide, is comprised the following steps：
(1) subcritical water is handled：0.3 kg abalones internal organ and 1.2 L pure water, the temperature of reactor are added in pyroreaction kettle
Degree and pressure are improved to 140 DEG C and 4.0 MPa rapidly, and room temperature simultaneously pressure release is cooled to after the min of heat-insulation pressure keeping 40, uses filtered through gauze
Obtain protein extracting liquid and filter residue；
(2) digest：Step (1) is obtained 4.5g flavor proteases and 1.5g pectases, enzyme are added in about 1.2L protein extracting liquid
3 h of solution reaction；Complex enzyme zymohydrolysis condition is pH 7.0, temperature 45 C.
(3) separate, concentrate and be spray-dried：12 g activated carbons are added in the protein enzymatic hydrolyzate that step (2) obtains,
15min is incubated at 45 DEG C, then removes activated carbon using disk centrifugal separator；Normal temperature is cooled to, pH is confirmed and adjusts to 7.0, lead to
Cross the mixed fillers (mixed proportion of the chelate fibre composition of macroporous absorbent resin, the chelating resin for adsorbing heavy metal and arsenic-adsorbing
For 7:2:1) column chromatography post decolourized, deodorant, removing heavy metal, dearsenification, residence time of the liquid in chromatographic column be 35
Min, it is 10% to carry out desalination and concentration to solid concentration using the NF membrane that molecular cut off is 150 Da, passes through vacuum concentration
It is 30% to solid concentration, finally spray drying is prepared into abalone proteins peptide powder；
(4) production：According to abalone proteins peptide powder 30%, beta-schardinger dextrin 60%, glue in the abalone proteins peptide powder of extraction
Former peptide 2%, soybean lecithin 5%, blue or green olive pulp extract 2.5%, the addition of honey 0.5% are well mixed, and particle is made, adds
3% magnesium stearate of granular mass, it is well mixed, is pressed into chewable tablets.
Embodiment 3
The present invention is a kind of method of subcritical water assistance enzymolysis extraction abalone proteins peptide, is comprised the following steps：
(1) subcritical water is handled：0.3 kg abalones internal organ and 1.5 L pure water, the temperature of reactor are added in pyroreaction kettle
Degree and pressure are improved to 160 DEG C and 1.0 MPa rapidly, and room temperature simultaneously pressure release is cooled to after the min of heat-insulation pressure keeping 15, uses filtered through gauze
Obtain protein extracting liquid and filter residue；
(2) digest：Step (1) is obtained 3.2g bromelains and 0.8g pectases, enzyme are added in about 1.4L protein extracting liquid
Solution reaction 2h；Complex enzyme zymohydrolysis condition is pH 6.0,40 DEG C of temperature.
(3) separate, concentrate and be spray-dried：28 g activated carbons are added in the protein enzymatic hydrolyzate that step (2) obtains,
30min is incubated at 40 DEG C, then removes activated carbon using disk centrifugal separator；Normal temperature is cooled to, pH is confirmed and adjusts to 6.0, lead to
Cross the mixed fillers (mixed proportion of the chelate fibre composition of macroporous absorbent resin, the chelating resin for adsorbing heavy metal and arsenic-adsorbing
For 7:2:1) column chromatography post decolourized, deodorant, removing heavy metal, dearsenification, residence time of the liquid in chromatographic column be 60
Min, it is 10% to carry out desalination and concentration to solid concentration using the NF membrane that molecular cut off is 150 Da, passes through vacuum concentration
It is 30% to solid concentration, finally spray drying is prepared into abalone proteins peptide powder；
(4) production：According to abalone proteins peptide powder 27%, beta-schardinger dextrin 55%, glue in the abalone proteins peptide powder of extraction
Former peptide 10%, soybean lecithin 4%, blue or green olive pulp extract 2%, the addition of honey 2% are well mixed, and particle is made, and add
5% magnesium stearate of grain quality, is well mixed, is pressed into chewable tablets.
2nd, product
The present invention is product made of a kind of subcritical water assistance enzymolysis extraction abalone proteins peptide, its constituent content by weight
For：Abalone proteins peptide powder 20% ~ 30%, beta-schardinger dextrin 55% ~ 70%, collagen peptide 2% ~ 10%, soybean lecithin 3% ~ 5%, blue or green olive
Pulp extract 1.0% ~ 2.5%, honey 0.5% ~ 2.0%.
It should be noted that abalone internal organ by after Subcritical Water Extraction albumen caused filter residue pass through after drying directly
It can be used as alga fertilizer.
The analysis of physical and chemical property of abalone proteins peptide produced by the invention：
Subcritical water has the H of higher concentration+And OH-, have to organic matters such as the albumen in abalone internal organ and polysaccharide stronger
Capacity of decomposition.The application enables the albumen of abalone internal organ to be dissolved out by control pressure and temperature, but does not digest complete
Dissolving, which will not occur, for marine alga to be removed by strainer filtering, and the filter residue can be made by drying directly as alga fertilizer
With discarded object will not be produced.The albumen of dissolving passes through egg in protease and the protein peptides of pectase progress complex enzyme zymohydrolysis preparation
Bai Hanliang is up to 65% or so, but also contains a small amount of polysaccharide (table 1), this be mainly abalone using ingest marine alga and planktonic organism as
It is raw, therefore in internal organ in addition to the albumen of rich content, also containing polysaccharide and its metabolite.To the abalone proteins peptide of preparation
Molecular weight is detected, the results showed that molecular weight of albumen is mainly distributed on the Da of 180 Da ~ 1000 in abalone peptide, and abundance is higher
Protein peptides are distributed in the Da (table 2) of 180 Da ~ 500, and the abalone proteins peptide for showing to obtain mainly is made up of 2-4 amino acid
Small peptide.It is absorbed by the body utilization it is reported that oligopeptides especially dipeptides or tripeptides easily pass through mucous membrane of small intestine.Therefore, the application system
Standby abalone proteins peptide is easily absorbed by the body, so as to play its bioactive functions.
The Hydroxyl radical-scavenging ability and DPPH radical scavenging activities of abalone proteins peptide are detected, as a result find it
IC50It is worth all very low (Fig. 1), can be close to typical antioxidation activity small peptide glutathione (Hydroxyl radical-scavenging ability
IC50Value=2.46；DPPH radical scavenging activities IC50Value=0.72), show that abalone proteins peptide prepared by the present invention has well
Antioxidation activity.According to influence (Fig. 2) of the abalone proteins peptide to breast cancer MDA-MB-231 cell inhibitory effects, it can be seen that
With the increase of abalone proteins peptide content in nutrient solution, the survival rate of breast cancer MDA-MB-231 cells is on a declining curve.Work as training
When the concentration of abalone proteins peptide is 4 mg/mL in nutrient solution, breast cancer MDA-MB-231 cell survival rates only have 40% ~ 50%.These
As a result show, abalone proteins peptide to the inhibited proliferation of breast cancer cell significantly (P<0.05), and be in dose dependent.
Subjective appreciation is carried out to the chewable tablets made using abalone proteins peptide, does not experience bitter taste and astringent taste, it is only fresh
Taste and sweet taste, show good in taste, will easily be esthetically acceptable to the consumers.
In summary, first abalone internal organ are carried out with processing extracting protein using subcritical water, recycles pectase and albumen
Enzyme is digested to the albumen of extraction, and enzymolysis liquid is concentrated, is concentrated in vacuo and is spray-dried by column chromatography for separation, NF membrane and prepared
Abalone proteins peptide powder, and abalone proteins peptide powder development function health product is utilized, to realize abalone internal organ higher value application
Reference is provided.
The specific embodiment of the present invention is above are only, but the design concept of the present invention is not limited thereto, it is all to utilize this structure
Think the change to present invention progress unsubstantiality, the behavior for invading the scope of the present invention all should be belonged to.
The basis of the abalone proteins peptide of table 1
| Embodiment 1 | Embodiment 2 | Embodiment 3 | |
| Protein content (%) | 65.45 | 63.11 | 67.85 |
| Polyoses content (%) | 14.90 | 15.31 | 12.09 |
| Moisture (%) | 7.12 | 8.36 | 7.02 |
The molecular weight distribution (%) of the basis of the abalone proteins peptide of table 2
| Molecular weight distribution (Da) | Embodiment 1 | Embodiment 2 | Embodiment 3 |
| ≥1000 | 6.95 | 6.08 | 6.45 |
| 500-1000 | 27.27 | 26.14 | 25.87 |
| 180-500 | 60.71 | 62.92 | 63.09 |
| ≤180 | 5.07 | 4.86 | 4.59 |
Claims (8)
- A kind of 1. method of subcritical water assistance enzymolysis extraction abalone proteins peptide, it is characterised in that：Comprise the following steps：(1) it is sub- Critical water process：Abalone internal organ add pure water by 13 ~ 15 (w/v) solid-liquid ratios, and 15 min are extracted under subcritical water condition ~ 60 min, it is cooled to after room temperature and obtains protein extracting liquid with stainless steel filter screen removing filter residue, filter residue after drying by that can make For alga fertilizer；(2) digest：Complex enzyme is added in the protein extracting liquid that step (1) obtains, complex enzyme addition extracts for albumen Take 0.1% ~ 0.5% (w/v) of liquid, the h of the h of enzyme digestion reaction 1 ~ 3；(3) separate, concentrate and be spray-dried：Step (2) is obtained It is dense by NF membrane after protein enzymatic hydrolyzate is decolourized using activated carbon and resin, deodorant, removing heavy metal, dearsenification etc. are refined Contracting, vacuum concentration and spray drying are prepared into abalone proteins peptide powder；(4) production：In the abalone proteins peptide powder of extraction Middle addition beta-schardinger dextrin, collagen peptide, soybean lecithin, blue or green olive pulp extract and honey, are well mixed, product are made Grain, 1% ~ 5% magnesium stearate of product particle quality is added, be well mixed, be pressed into chewable tablets.
- 2. the method for subcritical water assistance enzymolysis extraction abalone proteins peptide as claimed in claim 1, it is characterised in that：In step (1) in, described subcritical water refers to that temperature is 120 DEG C ~ 160 DEG C, and pressure is the MPa of 1.0 MPa ~ 7.0 water.
- 3. the method for subcritical water assistance enzymolysis extraction abalone proteins peptide as claimed in claim 1, it is characterised in that：In step (2) in, described complex enzyme refers to that papain, flavor protease, neutral proteinase and bromelain etc. are wherein appointed A kind of protease is compounded with pectase, and the compound proportion of protease and pectase is 3:1~5:1；.
- 4. the method for subcritical water assistance enzymolysis extraction abalone proteins peptide as claimed in claim 1, it is characterised in that：In step (2) in, the enzymatic hydrolysis condition of described complex enzyme is pH 5.0 ~ 7.0,40 DEG C ~ 50 DEG C of temperature.
- 5. the method for subcritical water assistance enzymolysis extraction abalone proteins peptide as claimed in claim 1, it is characterised in that：In step (3) in, described activated carbon decolorizing deodorant condition is：The pH of protein enzymatic hydrolyzate is adjusted to 5.0 ~ 6.0, and the addition of activated carbon is egg 0.5% ~ 2.0% (w/v) of white enzymolysis liquid volume, the min of 15 min ~ 60 is incubated at 40 DEG C ~ 50 DEG C, then utilizes disk centrifugal Machine removes activated carbon；Described resin deodorant removes heavy metal dearsenification condition：By the chela of macroporous absorbent resin, absorption heavy metal The mixed fillers of the chelate fibre of resin and arsenic-adsorbing composition, its mixed proportion are 7:2:1；The pH of protein enzymatic hydrolyzate be 5.0 ~ 6.0, protein enzymatic hydrolyzate residence time in mixed fillers is the min of 15 min ~ 30.
- 6. the method for subcritical water assistance enzymolysis extraction abalone proteins peptide as claimed in claim 1, it is characterised in that：In step (4) in, the constituent content of described product particle by weight is：Abalone proteins peptide powder 20% ~ 30%, beta-schardinger dextrin 55% ~ 70%th, collagen peptide 2% ~ 10%, soybean lecithin 3% ~ 5%, blue or green olive pulp extract 1.0% ~ 2.5%, honey 0.5% ~ 2.0%.
- 7. the method for subcritical water assistance enzymolysis extraction abalone proteins peptide as claimed in claim 1, it is characterised in that：Described Abalone internal organ are one kind in fresh abalone internal organ, frozen abalone internal organ or heat-treated abalone internal organ.
- 8. product made of a kind of subcritical water assistance enzymolysis extraction abalone proteins peptide according to claim 1, its feature It is：Its constituent content by weight is：Abalone proteins peptide powder 20% ~ 30%, beta-schardinger dextrin 55% ~ 70%, collagen peptide 2% ~ 10%th, soybean lecithin 3% ~ 5%, blue or green olive pulp extract 1.0% ~ 2.5%, honey 0.5% ~ 2.0%.
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