CN115725681A - Enzymatic extraction process and equipment for bioactive peptide - Google Patents
Enzymatic extraction process and equipment for bioactive peptide Download PDFInfo
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- CN115725681A CN115725681A CN202211470763.3A CN202211470763A CN115725681A CN 115725681 A CN115725681 A CN 115725681A CN 202211470763 A CN202211470763 A CN 202211470763A CN 115725681 A CN115725681 A CN 115725681A
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- 108090000765 processed proteins & peptides Proteins 0.000 title claims abstract description 61
- 230000000975 bioactive effect Effects 0.000 title claims abstract description 40
- 238000000605 extraction Methods 0.000 title claims abstract description 32
- 230000002255 enzymatic effect Effects 0.000 title claims abstract description 25
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 50
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 50
- 239000002994 raw material Substances 0.000 claims abstract description 50
- 102000004190 Enzymes Human genes 0.000 claims abstract description 33
- 108090000790 Enzymes Proteins 0.000 claims abstract description 33
- 230000007062 hydrolysis Effects 0.000 claims abstract description 28
- 238000006460 hydrolysis reaction Methods 0.000 claims abstract description 28
- 238000000034 method Methods 0.000 claims abstract description 21
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 21
- 238000002474 experimental method Methods 0.000 claims abstract description 19
- 238000012360 testing method Methods 0.000 claims abstract description 14
- 239000000758 substrate Substances 0.000 claims abstract description 5
- 230000000415 inactivating effect Effects 0.000 claims abstract description 4
- 229940088598 enzyme Drugs 0.000 claims description 28
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 22
- 230000009849 deactivation Effects 0.000 claims description 20
- 239000000287 crude extract Substances 0.000 claims description 16
- 238000009835 boiling Methods 0.000 claims description 15
- 238000010438 heat treatment Methods 0.000 claims description 15
- 239000003463 adsorbent Substances 0.000 claims description 12
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical group [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims description 10
- 108091005804 Peptidases Proteins 0.000 claims description 10
- 239000004365 Protease Substances 0.000 claims description 10
- 238000006243 chemical reaction Methods 0.000 claims description 9
- 102000035195 Peptidases Human genes 0.000 claims description 8
- 239000007921 spray Substances 0.000 claims description 6
- 239000006228 supernatant Substances 0.000 claims description 6
- 238000001179 sorption measurement Methods 0.000 claims description 5
- 230000001954 sterilising effect Effects 0.000 claims description 5
- 238000004040 coloring Methods 0.000 claims description 4
- 238000001035 drying Methods 0.000 claims description 4
- 230000003301 hydrolyzing effect Effects 0.000 claims description 4
- 238000005360 mashing Methods 0.000 claims description 4
- 239000007769 metal material Substances 0.000 claims description 4
- 239000000126 substance Substances 0.000 claims description 4
- 108010019160 Pancreatin Proteins 0.000 claims description 3
- 108090000526 Papain Proteins 0.000 claims description 3
- 108090000284 Pepsin A Proteins 0.000 claims description 3
- 102000057297 Pepsin A Human genes 0.000 claims description 3
- 102000004142 Trypsin Human genes 0.000 claims description 3
- 108090000631 Trypsin Proteins 0.000 claims description 3
- 238000005119 centrifugation Methods 0.000 claims description 3
- 238000013461 design Methods 0.000 claims description 3
- 239000007788 liquid Substances 0.000 claims description 3
- 229940055695 pancreatin Drugs 0.000 claims description 3
- 229940055729 papain Drugs 0.000 claims description 3
- 235000019834 papain Nutrition 0.000 claims description 3
- 229940111202 pepsin Drugs 0.000 claims description 3
- 238000012545 processing Methods 0.000 claims description 3
- 239000000047 product Substances 0.000 claims description 3
- 235000019833 protease Nutrition 0.000 claims description 3
- 238000000746 purification Methods 0.000 claims description 3
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- 239000012588 trypsin Substances 0.000 claims description 3
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- 108090000145 Bacillolysin Proteins 0.000 claims 1
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- 235000019658 bitter taste Nutrition 0.000 abstract 1
- 230000000694 effects Effects 0.000 abstract 1
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- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 description 2
- 108090000787 Subtilisin Proteins 0.000 description 2
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- 150000001413 amino acids Chemical class 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 239000012467 final product Substances 0.000 description 2
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Abstract
The invention discloses an enzymatic extraction process and equipment of bioactive peptide, which comprises the following steps: s1, preprocessing; s2, carrying out comparison test; s3, performing enzymolysis; s4, inactivating enzyme; s5, separating and purifying; s6, removing bitter taste and taking color; s7, subsequent treatment, and relates to the technical field of enzymatic extraction processes of bioactive peptides. The enzymatic extraction process and equipment for the bioactive peptide are characterized in that comparison experiment steps are set, after raw material extraction of protein is completed, small parts of the protein are taken out and respectively put into a plurality of experiment test tubes in a quantitative mode, hydrolysis effects of a plurality of samples are compared through the variable substrate concentration, the enzyme concentration, the PH value and the temperature, and therefore the optimal hydrolysis condition is selected, the process can be adapted to various crops or plants, the process can be carried out under the optimal hydrolysis condition when the bioactive peptide is extracted from the various crops or plants, the efficiency of the whole hydrolysis process of the process is guaranteed, and the efficiency of the whole extraction process is improved.
Description
Technical Field
The invention relates to the technical field of an enzymatic extraction process of bioactive peptides, in particular to an enzymatic extraction process of bioactive peptides and equipment thereof.
Background
A peptide is an organic compound consisting of amino acids linked together by peptide bonds, typically peptides consisting of more than 10 amino acids are called polypeptides. The polypeptide has a wide variety and wide distribution range in human bodies, is not only an important component and nutrient substance of the human bodies, but also shows bioactivity on protein synthesis, cell function regulation, immunoregulation, metabolic growth and development and the like of the human bodies. On the other hand, most proteins are not soluble in acidic beverages, however, active protein peptides are sufficiently soluble in acidic beverages and are low in concentration and not greasy to the mouth. How to develop novel polypeptide beverage is a research hotspot of health care beverage, sports beverage, functional beverage and the like.
The existing preparation method of polypeptide beverage mainly can only extract biological peptide from one crop, for example, chinese patent (CN 104256828B) discloses a preparation method of wheat germ polypeptide beverage, and for example, chinese patent (CN 201010534301.4) discloses a preparation method of walnut polypeptide beverage, and the existing preparation method of polypeptide beverage lacks a process or method capable of extracting biological peptide from various crops or plants, and can not meet social and market requirements.
Disclosure of Invention
Aiming at the defects of the prior art, the invention provides an enzymatic extraction process and equipment of bioactive peptides, and solves the problem that the prior process or method capable of extracting bioactive peptides of various crops or plants is lacked and cannot meet the requirements of society and market.
In order to realize the purpose, the invention is realized by the following technical scheme: an enzymatic extraction process of bioactive peptide comprises the following steps:
s1, pretreatment: placing the raw materials to be extracted into a container, adding boiling water, boiling for 10-60min, mashing the rest sample, centrifuging, and collecting the precipitated protein raw material;
s2, comparison experiment: selecting a part of protein raw materials, and determining the optimal substrate concentration, enzyme concentration, pH value, temperature and the selection type of protein enzyme during protein enzymolysis by using the part of protein raw materials in an orthogonal design experiment mode;
s3, enzymolysis: putting the rest protein raw materials into a container, pouring the determined enzyme for reaction in proportion, adjusting the pH value and the temperature to appropriate states, and hydrolyzing the protein raw materials; s4, enzyme deactivation: pouring the protein raw material after the enzymolysis into boiling water at 100 ℃ for water bath, inactivating various proteases for reaction, and stopping the reaction to continue to avoid further hydrolysis of the peptide; s5, separation and purification: centrifuging the enzymolysis liquid at high speed, and removing unconverted protein and other insoluble substances to obtain crude extract of bioactive peptide; s6, debitterizing and coloring: adding an adsorbent which accounts for 10 to 20 percent of the weight of the crude extract of the bioactive peptide into the crude extract of the bioactive peptide, uniformly stirring, adsorbing for 1 to 3 hours, and taking supernatant; s7, subsequent processing: and sterilizing, concentrating and drying the supernatant to obtain a finished product.
Further, the plurality of said proteinases include pepsin, trypsin, pancreatin, papain and subtilisin, and optionally one or more thereof.
Further, the water bath time of the protein raw material is 5-10min.
Further, the adsorbent is an activated carbon adsorbent.
An enzymatic extraction device for bioactive peptides comprises a heating tank, a first centrifuge, a plurality of experimental test tubes, a hydrolysis device, a second centrifuge, an adsorption tank and a spray dryer; the hydrolysis device comprises a base, a plurality of support rods of the vertical fixedly connected with of upper surface of the base, the top fixedly connected with heating device of base, it is a plurality of just be located between the bracing piece the board is placed to the horizontal fixedly connected with in heating device's top, the top detachably of placing the board installs enzyme deactivation jar, and is a plurality of fixedly connected with go-between the top of bracing piece, the inside fixedly connected with enzymolysis tank of go-between, the bottom intercommunication of enzymolysis tank has communicating pipe, the bottom of communicating pipe extends to the inside of enzyme deactivation jar.
Further, a thermometer is arranged inside the enzyme deactivation tank.
Further, an electromagnetic valve is arranged on the communicating pipe.
Further, the placing plate is made of a metal material having good thermal conductivity.
Compared with the prior art, the invention has the beneficial effects that:
the enzymatic extraction process and equipment for the bioactive peptide are characterized in that a comparison experiment step is added between the pretreatment process of protein and the protein enzymolysis process, after the extraction of the raw material of the protein is completed, small parts of the raw material of the protein are taken out and respectively put into a plurality of experiment test tubes in a quantitative mode, different proteases are selected to be respectively sent into different experiment test tubes in different concentrations, then the hydrolysis states of the protein in the experiment test tubes are observed under different PH values and different temperature states, the most suitable conditions of the current protein raw material can be determined after the comparison of a plurality of samples, the process can be adapted to various crops or plants, and the extraction of the bioactive peptide from various crops or plants can be carried out under the best hydrolysis conditions, so that the efficiency of the whole hydrolysis process of the process is ensured, and the efficiency of the whole extraction process is improved.
Drawings
FIG. 1 is a flow chart of the process for enzymatic extraction of bioactive peptides according to the present invention;
FIG. 2 is a schematic diagram of an enzymatic extraction apparatus for bioactive peptides according to the present invention;
FIG. 3 is a schematic view of the hydrolysis apparatus of the present invention.
In the figure: 1. a heating tank; 2. a first centrifuge; 3. an experimental test tube; 4. a hydrolysis device; 41. a base; 42. a support bar; 43. a heating device; 44. placing the plate; 45. an enzyme deactivation tank; 46. a thermometer; 47. a connecting ring; 48. an enzymolysis tank; 49. a communicating pipe; 410. an electromagnetic valve; 5. a second centrifuge; 6. an adsorption tank; 7. an activated carbon adsorbent; 8. a spray dryer.
Detailed Description
The technical solutions in the embodiments of the present invention will be clearly and completely described below with reference to the drawings in the embodiments of the present invention, and it is obvious that the described embodiments are only a part of the embodiments of the present invention, and not all of the embodiments. All other embodiments, which can be derived by a person skilled in the art from the embodiments given herein without making any creative effort, shall fall within the protection scope of the present invention.
Referring to fig. 1-3, the present invention provides a technical solution: an enzymatic extraction process of bioactive peptide comprises the following steps:
s1, pretreatment: placing the raw materials to be extracted into a container, adding boiling water, boiling for 10-60min, mashing the rest sample, centrifuging, and collecting the precipitated protein raw material;
s2, comparison experiment: selecting a part of protein raw materials, and determining the optimal substrate concentration, enzyme concentration, pH value, temperature and the selection type of protein enzyme during protein enzymolysis by using the part of protein raw materials in an orthogonal design experiment mode;
s3, enzymolysis: putting the rest protein raw materials into a container, pouring the determined enzyme for reaction in proportion, adjusting the pH value and the temperature to appropriate states, and hydrolyzing the protein raw materials; s4, enzyme deactivation: pouring the protein raw material after the enzymolysis into boiling water at 100 ℃ for water bath, inactivating various proteases for reaction, and stopping the reaction to continue to avoid further hydrolysis of the peptide; s5, separation and purification: centrifuging the enzymolysis liquid at high speed, and removing unconverted protein and other insoluble substances to obtain crude extract of bioactive peptide; s6, debitterizing and coloring: adding an adsorbent which accounts for 10-20% of the weight of the crude extract of the bioactive peptide into the crude extract of the bioactive peptide, uniformly stirring, adsorbing for 1-3 hours, and taking supernatant; s7, subsequent processing: and sterilizing, concentrating and drying the supernatant to obtain a finished product.
Part of the raw materials can be directly used as protein raw materials after being boiled and mashed in boiling water, and a centrifugal link is not needed.
The protease comprises pepsin, trypsin, pancreatin, papain and subtilisin, and one or more of them can be selected.
The water bath time of the protein raw material is 5-10min.
The adsorbent is an activated carbon adsorbent 7.
An enzymatic extraction device of bioactive peptide comprises a heating tank 1, a first centrifuge 2, a plurality of experiment test tubes 3, a hydrolysis device 4, a second centrifuge 5, an adsorption tank 6 and a spray dryer 8.
When the raw materials are pretreated, water is poured into the heating tank 1 and heated to boil the raw materials, the boiling time is different within 10-60min according to the difference of the raw materials, after the raw materials are completely boiled, the raw materials are taken out, smashed and poured into water 4-10 times of the volume of the raw materials for mixing, after the mixing is finished, the raw materials are centrifuged by the first centrifuge 2, and after the centrifugation is finished, the precipitate is collected to obtain the protein raw material.
In the comparison test, a small part of protein raw materials are extracted and are respectively placed into different experiment test tubes 3, different proteases are added into each experiment test tube 3, then the pH value and the temperature are controlled to observe the hydrolysis state of the proteins in each experiment test tube 3, and the comparison test is carried out for a plurality of times by changing the substrate concentration, the protease concentration, the pH value and the temperature, so that the most hydrolysis condition of the current proteins is obtained, the process can be adapted to various crops or plants, and the process can be carried out under the most hydrolysis condition when active peptide extraction is carried out on various crops or plants.
The hydrolysis unit includes base 41, a plurality of bracing pieces 42 of the vertical fixedly connected with of upper surface of base 41, base 41's top fixedly connected with heating device 43, the board 44 is placed to the horizontal fixedly connected with in top that just is located heating device 43 between a plurality of bracing pieces 42, the enzyme jar 45 that goes out is installed to the top detachably of placing board 44, fixedly connected with go-between 47 between a plurality of bracing pieces 42's the top, go-between 47's inside fixedly connected with enzymolysis tank 48, the bottom intercommunication of enzymolysis tank 48 has communicating pipe 49, the bottom of communicating pipe 49 extends to the inside of enzyme jar 45 that goes out.
The communication pipe 49 is provided with an electromagnetic valve 410.
After the optimal hydrolysis conditions are determined through a comparison experiment, the hydrolysis device 4 can be used for carrying out hydrolysis operation on the residual protein raw materials, the protein raw materials enter the enzyme deactivation tank 45 through the communicating pipe 49 after hydrolysis in the enzymolysis tank 48 is completed, water is filled in the enzyme deactivation tank 45 in advance, the electromagnetic valve 410 can be opened after the water in the enzyme deactivation tank 45 is heated to boiling through the heating device 43, and the hydrolyzed protein raw materials enter the enzyme deactivation tank 45 for carrying out enzyme deactivation operation.
The enzyme deactivation pot 45 is provided with a thermometer 46 inside.
The placement plate 44 is made of a metal material having good thermal conductivity.
The thermometer 46 is convenient for people to determine the temperature of the water in the enzyme deactivation tank 45, and ensures that the enzyme deactivation operation can be successfully completed.
The placing plate 44 is made of a metal material with good thermal conductivity, so that heat emitted by the heating device 43 can quickly act on water in the enzyme deactivation tank 45, and the working efficiency is improved.
After enzyme deactivation, a second centrifuge 5 is used for centrifugation to obtain crude extract of active peptide, and then the crude extract is sent into an adsorption tank 6, and an active carbon adsorbent 7 with the mass of 10% -20% of the crude extract is poured into the crude extract to carry out debittering and color extraction on the crude extract to obtain refined active peptide solution.
The refined active peptide solution is subjected to ultrahigh temperature instant sterilization at 135 ℃ for 5min, then concentrated and dried by a spray dryer 8 to obtain a final product.
The principle of the enzymatic extraction process and the equipment of the bioactive peptide provided by the invention is as follows: firstly, boiling raw materials by using a heating tank 1, after the boiling is finished, mashing the raw materials, pouring the mashed raw materials into water with the volume 4-10 times of that of the raw materials, mixing the mixture with the water, centrifuging the mixture by using a first centrifuge 2 to obtain a protein raw material, then obtaining the optimal hydrolysis condition of the current protein by using a comparison experiment, hydrolyzing the protein raw material by using a hydrolysis device 4 under the condition, centrifuging the protein mixed solution after the hydrolysis is finished by using a second centrifuge 5 to obtain a bioactive peptide crude extract, then debittering and coloring the crude extract by using an active carbon adsorbent 7 to obtain a refined bioactive peptide solution, finally, instantly sterilizing the refined bioactive peptide solution at the high temperature of 135 ℃ for 5min, concentrating the sterilized active peptide solution, and drying the concentrated active peptide solution by using a spray dryer 8 to obtain a final product.
It is noted that, herein, relational terms such as first and second, and the like may be used solely to distinguish one entity or action from another entity or action without necessarily requiring or implying any actual such relationship or order between such entities or actions. Also, the terms "comprises," "comprising," or any other variation thereof, are intended to cover a non-exclusive inclusion, such that a process, method, article, or apparatus that comprises a list of elements does not include only those elements but may include other elements not expressly listed or inherent to such process, method, article, or apparatus.
The above description is only for the purpose of illustrating the preferred embodiments of the present invention and is not to be construed as limiting the invention, and any modifications, equivalents, improvements and the like that fall within the spirit and principle of the present invention are intended to be included therein.
Claims (9)
1. An enzymatic extraction process of bioactive peptide is characterized in that: the method comprises the following steps:
s1, pretreatment: placing the raw materials to be extracted into a container, adding boiling water, boiling for 10-60min, mashing the rest sample, centrifuging, and collecting the precipitated protein raw material;
s2, comparison experiment: selecting a part of protein raw materials, and determining the optimal substrate concentration, enzyme concentration, pH value, temperature and the selection type of protein enzyme during protein enzymolysis by using the part of protein raw materials in an orthogonal design experiment mode;
s3, enzymolysis: putting the rest protein raw materials into a container, pouring the determined enzyme for reaction in proportion, adjusting the pH value and the temperature to appropriate states, and hydrolyzing the protein raw materials; s4, enzyme deactivation: pouring the protein raw material after the enzymolysis into boiling water at 100 ℃ for water bath, inactivating various proteases for reaction, and stopping the reaction to continue to avoid further hydrolysis of the peptide; s5, separation and purification: centrifuging the enzymolysis liquid at high speed, and removing unconverted protein and other insoluble substances to obtain crude extract of bioactive peptide; s6, debitterizing and coloring: adding an adsorbent which accounts for 10 to 20 percent of the weight of the crude extract of the bioactive peptide into the crude extract of the bioactive peptide, uniformly stirring, adsorbing for 1 to 3 hours, and taking supernatant; s7, subsequent processing: and sterilizing, concentrating and drying the supernatant to obtain a finished product.
2. The enzymatic extraction process of bioactive peptide according to claim 1, wherein: in the S1, part of the raw materials can be directly used as the protein raw materials after being boiled and smashed in boiling water, and a centrifugation link is not needed.
3. The enzymatic extraction process and equipment of bioactive peptides as claimed in claim 1, wherein: in the S2, the plurality of proteinases comprise pepsin, trypsin, pancreatin, papain and bacillus subtilis neutral protease, and one or more proteinases can be selected.
4. The enzymatic extraction process and apparatus of bioactive peptide as claimed in claim 1, wherein: in S4, the water bath time of the protein raw material is 5-10min.
5. The enzymatic extraction process and apparatus of bioactive peptide as claimed in claim 1, wherein: in the step S6, the adsorbent is an activated carbon adsorbent (7).
6. An enzymatic extraction device of bioactive peptide is characterized in that: comprises a heating tank (1), a first centrifuge (2), a plurality of experiment test tubes (3), a hydrolysis device (4), a second centrifuge (5), an adsorption tank (6) and a spray dryer (8); the hydrolysis device comprises a base (41), a plurality of supporting rods (42) of the vertical fixedly connected with of upper surface of base (41), top fixedly connected with heating device (43) of base (41), it is a plurality of just be located between supporting rod (42) board (44) is placed to the horizontal fixedly connected with in top of heating device (43), the top detachably of placing board (44) installs enzyme deactivation jar (45), and is a plurality of fixedly connected with go-between (47) between the top of supporting rod (42), the inside fixedly connected with enzymolysis tank (48) of go-between (47), the bottom intercommunication of enzymolysis tank (48) has communicating pipe (49), the bottom of communicating pipe (49) extends to the inside of enzyme deactivation jar (45).
7. The enzymatic extraction apparatus of bioactive peptides as claimed in claim 5, wherein: a thermometer (46) is arranged in the enzyme deactivation tank (45).
8. The enzymatic extraction device of bioactive peptides as claimed in claim 5, wherein: an electromagnetic valve (410) is arranged on the communicating pipe (49).
9. The enzymatic extraction device of bioactive peptides as claimed in claim 5, wherein: the placement plate (44) is made of a metal material having good thermal conductivity.
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| CN202211470763.3A CN115725681A (en) | 2022-11-23 | 2022-11-23 | Enzymatic extraction process and equipment for bioactive peptide |
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| CN202211470763.3A CN115725681A (en) | 2022-11-23 | 2022-11-23 | Enzymatic extraction process and equipment for bioactive peptide |
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Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120302731A1 (en) * | 2010-01-14 | 2012-11-29 | South China University Of Technology | Protein hydrolysate, polypeptide solution and polypeptide, preparation method and use thereof |
| CN106350561A (en) * | 2016-11-11 | 2017-01-25 | 六盘水师范学院 | Tartary buckwheat active peptide and extraction method thereof |
| CN107190038A (en) * | 2016-03-14 | 2017-09-22 | 南通仁寿食品有限公司 | Microorganism prepares active peptide technique |
| US20180023110A1 (en) * | 2015-01-30 | 2018-01-25 | Jiangsu University | Method for preparing functional polypeptide through multimode ultrasonic enhancing enzymolysis |
| CN212199287U (en) * | 2020-04-09 | 2020-12-22 | 厦门元之道生物科技有限公司 | Collagen peptide low temperature enzymolysis equipment |
-
2022
- 2022-11-23 CN CN202211470763.3A patent/CN115725681A/en active Pending
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| US20120302731A1 (en) * | 2010-01-14 | 2012-11-29 | South China University Of Technology | Protein hydrolysate, polypeptide solution and polypeptide, preparation method and use thereof |
| US20180023110A1 (en) * | 2015-01-30 | 2018-01-25 | Jiangsu University | Method for preparing functional polypeptide through multimode ultrasonic enhancing enzymolysis |
| CN107190038A (en) * | 2016-03-14 | 2017-09-22 | 南通仁寿食品有限公司 | Microorganism prepares active peptide technique |
| CN106350561A (en) * | 2016-11-11 | 2017-01-25 | 六盘水师范学院 | Tartary buckwheat active peptide and extraction method thereof |
| CN212199287U (en) * | 2020-04-09 | 2020-12-22 | 厦门元之道生物科技有限公司 | Collagen peptide low temperature enzymolysis equipment |
Non-Patent Citations (2)
| Title |
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| SUSY PIOVESANA等: "Recent trends and analytical challenges in plant bioactive peptide separation, identification and validation", 《ANALYTICAL AND BIOANALYTICAL CHEMISTRY》, vol. 410, 20 January 2018 (2018-01-20), pages 3425, XP036509427, DOI: 10.1007/s00216-018-0852-x * |
| 刘公博等: "酶解法制备大豆肽工艺的优化及苦味测定", 《青岛农业大学学报(自然科学版)》, vol. 37, no. 2, 31 December 2020 (2020-12-31), pages 123 - 128 * |
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