CN113667708A - Preparation method of plant intercellular substance - Google Patents
Preparation method of plant intercellular substance Download PDFInfo
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- CN113667708A CN113667708A CN202110967579.9A CN202110967579A CN113667708A CN 113667708 A CN113667708 A CN 113667708A CN 202110967579 A CN202110967579 A CN 202110967579A CN 113667708 A CN113667708 A CN 113667708A
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- protein
- intercellular substance
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- 239000000126 substance Substances 0.000 title claims abstract description 36
- 238000002360 preparation method Methods 0.000 title abstract description 8
- 241000196324 Embryophyta Species 0.000 claims abstract description 24
- 238000000855 fermentation Methods 0.000 claims abstract description 23
- 108090000623 proteins and genes Proteins 0.000 claims abstract description 21
- 102000004169 proteins and genes Human genes 0.000 claims abstract description 21
- 230000004151 fermentation Effects 0.000 claims abstract description 16
- 238000001914 filtration Methods 0.000 claims abstract description 16
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 claims abstract description 14
- 238000000034 method Methods 0.000 claims abstract description 14
- 241000220317 Rosa Species 0.000 claims abstract description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims abstract description 10
- 238000001556 precipitation Methods 0.000 claims abstract description 8
- 101710132601 Capsid protein Proteins 0.000 claims abstract description 7
- 238000004090 dissolution Methods 0.000 claims abstract description 7
- 239000000049 pigment Substances 0.000 claims abstract description 7
- 229920001184 polypeptide Polymers 0.000 claims abstract description 7
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 7
- 102000004196 processed proteins & peptides Human genes 0.000 claims abstract description 7
- 238000002156 mixing Methods 0.000 claims abstract description 6
- 230000001954 sterilising effect Effects 0.000 claims description 9
- 239000008213 purified water Substances 0.000 claims description 8
- 239000002244 precipitate Substances 0.000 claims description 5
- 238000004659 sterilization and disinfection Methods 0.000 claims description 5
- 241000628997 Flos Species 0.000 claims description 4
- 239000008055 phosphate buffer solution Substances 0.000 claims description 4
- 239000006228 supernatant Substances 0.000 claims description 4
- 239000011148 porous material Substances 0.000 claims description 3
- MCSXGCZMEPXKIW-UHFFFAOYSA-N 3-hydroxy-4-[(4-methyl-2-nitrophenyl)diazenyl]-N-(3-nitrophenyl)naphthalene-2-carboxamide Chemical compound Cc1ccc(N=Nc2c(O)c(cc3ccccc23)C(=O)Nc2cccc(c2)[N+]([O-])=O)c(c1)[N+]([O-])=O MCSXGCZMEPXKIW-UHFFFAOYSA-N 0.000 claims description 2
- 241000894006 Bacteria Species 0.000 abstract description 2
- 230000004071 biological effect Effects 0.000 abstract description 2
- 238000011031 large-scale manufacturing process Methods 0.000 abstract description 2
- 230000008569 process Effects 0.000 abstract description 2
- 239000000047 product Substances 0.000 description 5
- 238000000605 extraction Methods 0.000 description 4
- 239000012530 fluid Substances 0.000 description 3
- 230000000694 effects Effects 0.000 description 2
- 230000006870 function Effects 0.000 description 2
- 108010023832 Florigen Proteins 0.000 description 1
- 229910019142 PO4 Inorganic materials 0.000 description 1
- 241000109329 Rosa xanthina Species 0.000 description 1
- 235000004789 Rosa xanthina Nutrition 0.000 description 1
- 239000006177 biological buffer Substances 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 230000007547 defect Effects 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 230000001261 florigenic effect Effects 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 238000005374 membrane filtration Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- NBIIXXVUZAFLBC-UHFFFAOYSA-K phosphate Chemical compound [O-]P([O-])([O-])=O NBIIXXVUZAFLBC-UHFFFAOYSA-K 0.000 description 1
- 239000010452 phosphate Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 238000001179 sorption measurement Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P21/00—Preparation of peptides or proteins
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/145—Extraction; Separation; Purification by extraction or solubilisation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/30—Extraction; Separation; Purification by precipitation
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/34—Extraction; Separation; Purification by filtration, ultrafiltration or reverse osmosis
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K1/00—General methods for the preparation of peptides, i.e. processes for the organic chemical preparation of peptides or proteins of any length
- C07K1/14—Extraction; Separation; Purification
- C07K1/36—Extraction; Separation; Purification by a combination of two or more processes of different types
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- Chemical & Material Sciences (AREA)
- Organic Chemistry (AREA)
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Molecular Biology (AREA)
- Biochemistry (AREA)
- General Health & Medical Sciences (AREA)
- Genetics & Genomics (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Medicinal Chemistry (AREA)
- Biophysics (AREA)
- Analytical Chemistry (AREA)
- Engineering & Computer Science (AREA)
- Zoology (AREA)
- Wood Science & Technology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Microbiology (AREA)
- Biotechnology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Engineering & Computer Science (AREA)
- Water Supply & Treatment (AREA)
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention provides a preparation method of a plant intercellular substance. Mixing rose and water, covering with white oil, performing anaerobic fermentation, sequentially filtering, centrifuging and clarifying a fermentation product, and adding activated carbon to remove pigments; and then adjusting pH to precipitate polypeptide and protein substances, centrifuging and dissolving, purifying and concentrating the core protein in a continuous precipitation and dissolution mode, finally collecting high-concentration plant functional protein, and filtering to remove bacteria to obtain a product. The invention successfully prepares the high-purity plant intercellular substance and has higher biological activity. The method has the advantages of high preparation efficiency, simple process flow, easy large-scale production and developed technical advantages.
Description
Technical Field
The invention relates to the technical field of bioengineering, in particular to a preparation method of a plant intercellular substance.
Background
The intercellular substance is a substance which is produced by cells and has no cell morphology and structure, and comprises fibers, matrix and fluid substances, and the cells in the organism tissue are infiltrated in the intercellular substance, play supporting, protecting, connecting and nourishing roles to the cells and participate in forming the microenvironment for the survival of the cells. The intercellular substance is a fluid environment in which the cells are located, and the metabolites of the cells and the unused substances are received by the intercellular substance, and the cells and the intercellular substance are continuously exchanged, and the intercellular substance contains all substances required by the cells during metabolism.
The rose is the most famous and popular one of all flowers, has definite skin care effect besides the fragrance smell, and therefore, the rose extract is widely applied to skin care products. In the prior art, the conventional extraction method takes the whole rose flowers as extraction objects, and is difficult to accurately extract specific tissue fluid; in recent years, research shows that the rose intercellular substance has more outstanding skin care effect and has milder property compared with the intracellular components; under the condition, if the rose intercellular substance can be extracted specially, the efficacy of the related skin care product is expected to be improved, and the product value is improved.
Disclosure of Invention
The invention aims to provide a preparation method of plant intercellular substance aiming at the technical defects of the prior art, so as to solve the technical problem that a special extraction method of rose intercellular substance is lacked at present.
The invention also aims to solve the technical problem of how to fully ensure the extraction efficiency of the rose intercellular substance.
In order to achieve the technical purpose, the invention adopts the following technical scheme:
a method for preparing plant intercellular substance comprises the following steps: mixing flos Rosae Rugosae and purified water, adding fermentation strain, covering with white oil, performing anaerobic fermentation, fermenting for 3d, filtering, centrifuging, and clarifying; adding activated carbon to remove pigment, adjusting pH to precipitate polypeptide and protein, centrifuging supernatant at a centrifugal force of 12000G or more, collecting protein precipitate, and dissolving in phosphate buffer solution; repeating precipitation and dissolution, concentrating the core protein, collecting the concentrated protein, filtering and sterilizing to obtain the plant intercellular substance.
Preferably, the weight ratio of the rose to the purified water is 1: 2.
Preferably, the fermentation broth is dark red after 3 days of fermentation.
Preferably, the filter sterilization is performed using a filter with a pore size of 0.22 μm.
Preferably, the method further comprises the following steps: after filtration sterilization, it is added to the sterilized fermentation broth.
The invention provides a preparation method of a plant intercellular substance. Mixing rose and water, covering with white oil, performing anaerobic fermentation, sequentially filtering, centrifuging and clarifying a fermentation product, and adding activated carbon to remove pigments; and then adjusting pH to precipitate polypeptide and protein substances, centrifuging and dissolving, purifying and concentrating the core protein in a continuous precipitation and dissolution mode, finally collecting high-concentration plant functional protein, and filtering to remove bacteria to obtain a product. The invention successfully prepares the high-purity plant intercellular substance and has higher biological activity. The method has the advantages of high preparation efficiency, simple process flow, easy large-scale production and developed technical advantages.
Detailed Description
Hereinafter, specific embodiments of the present invention will be described in detail. Well-known structures or functions may not be described in detail in the following embodiments in order to avoid unnecessarily obscuring the details. Approximating language, as used herein in the following examples, may be applied to identify quantitative representations that could permissibly vary in number without resulting in a change in the basic function. Unless defined otherwise, technical and scientific terms used in the following examples have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs.
Example 1
A method for preparing plant intercellular substance comprises the following steps: adding purified water and fermentation strain into 25 tons of roses according to the ratio of 1:2, covering a layer of white oil outside to promote anaerobic fermentation, and after 3 days, making the fermentation liquor turn into deep red, and then filtering, centrifuging and clarifying. Removing pigment by activated carbon adsorption, promoting polypeptide protein substance precipitation by adjusting pH, centrifuging at over 12000G, collecting protein precipitate, and dissolving in phosphate biological buffer solution. The core protein is concentrated by continuous precipitation, dissolution and purification, and finally the plant functional protein with high concentration is collected, and the plant functional protein can be added into the sterilized fermentation liquor after membrane filtration of 0.22 mu m, the protein which is not used in time can be preserved, and the concentrated plant core raw material is the plant florigen or the plant living cell element (namely intercellular substance).
Example 2
A method for preparing plant intercellular substance comprises the following steps: mixing flos Rosae Rugosae and purified water, adding fermentation strain, covering with white oil, performing anaerobic fermentation, fermenting for 3d, filtering, centrifuging, and clarifying; adding activated carbon to remove pigment, adjusting pH to precipitate polypeptide and protein, centrifuging supernatant at a centrifugal force of 12000G or more, collecting protein precipitate, and dissolving in phosphate buffer solution; repeating precipitation and dissolution, concentrating the core protein, collecting the concentrated protein, filtering and sterilizing to obtain the plant intercellular substance. Wherein the weight ratio of the rose to the purified water is 1: 2. After fermenting for 3d, the fermentation liquor is dark red. The filter sterilization is realized by using a filter membrane with the pore size of 0.22 mu m.
Example 3
A method for preparing plant intercellular substance comprises the following steps: mixing flos Rosae Rugosae and purified water, adding fermentation strain, covering with white oil, performing anaerobic fermentation, fermenting for 3d, filtering, centrifuging, and clarifying; adding activated carbon to remove pigment, adjusting pH to precipitate polypeptide and protein, centrifuging supernatant at a centrifugal force of 12000G or more, collecting protein precipitate, and dissolving in phosphate buffer solution; repeating precipitation and dissolution, concentrating the core protein, collecting the concentrated protein, filtering and sterilizing to obtain the plant intercellular substance.
The embodiments of the present invention have been described in detail, but the 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. Any modification, equivalent replacement, and improvement made within the scope of the application of the present invention should be included in the protection scope of the present invention.
Claims (5)
1. A method for preparing a plant intercellular substance, comprising the steps of: mixing flos Rosae Rugosae and purified water, adding fermentation strain, covering with white oil, performing anaerobic fermentation, fermenting for 3d, filtering, centrifuging, and clarifying; adding activated carbon to remove pigment, adjusting pH to precipitate polypeptide and protein, centrifuging supernatant at a centrifugal force of 12000G or more, collecting protein precipitate, and dissolving in phosphate buffer solution; repeating precipitation and dissolution, concentrating the core protein, collecting the concentrated protein, filtering and sterilizing to obtain the plant intercellular substance.
2. The method for preparing a plant intercellular substance according to claim 1, wherein the weight ratio of rose flower to purified water is 1: 2.
3. The method according to claim 1, wherein the fermentation broth is deep red after 3 days of fermentation.
4. The method according to claim 1, wherein said filtration sterilization is performed using a 0.22 μm-pore filter.
5. The method of claim 1, further comprising the steps of: after filtration sterilization, it is added to the sterilized fermentation broth.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110967579.9A CN113667708A (en) | 2021-08-23 | 2021-08-23 | Preparation method of plant intercellular substance |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202110967579.9A CN113667708A (en) | 2021-08-23 | 2021-08-23 | Preparation method of plant intercellular substance |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN113667708A true CN113667708A (en) | 2021-11-19 |
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Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202110967579.9A Pending CN113667708A (en) | 2021-08-23 | 2021-08-23 | Preparation method of plant intercellular substance |
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| Country | Link |
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| CN (1) | CN113667708A (en) |
Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103446022A (en) * | 2013-09-02 | 2013-12-18 | 广州天玺生物科技有限公司 | Plant polypeptide protein extract and use thereof |
| CN107523599A (en) * | 2017-09-01 | 2017-12-29 | 兰溪市沉默生物科技有限公司 | The method that fermentation method prepares peony pollen polypeptide |
| WO2018100866A1 (en) * | 2016-11-30 | 2018-06-07 | キリン株式会社 | Method for producing useful protein by means of plant |
| CN110157758A (en) * | 2017-12-26 | 2019-08-23 | 浦江县欧立生物技术有限公司 | The purposes of the Flos Rosae Rugosas pollen polypeptide of fermentation method preparation |
| CN111588758A (en) * | 2020-07-07 | 2020-08-28 | 西藏云朵玫瑰科技有限公司 | Extraction method of rose extract |
| CN114457136A (en) * | 2022-01-27 | 2022-05-10 | 美尔健（深圳）生物科技有限公司 | Fermentation liquid based on rose fermentation recombinant collagen and application thereof |
-
2021
- 2021-08-23 CN CN202110967579.9A patent/CN113667708A/en active Pending
Patent Citations (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103446022A (en) * | 2013-09-02 | 2013-12-18 | 广州天玺生物科技有限公司 | Plant polypeptide protein extract and use thereof |
| WO2018100866A1 (en) * | 2016-11-30 | 2018-06-07 | キリン株式会社 | Method for producing useful protein by means of plant |
| CN107523599A (en) * | 2017-09-01 | 2017-12-29 | 兰溪市沉默生物科技有限公司 | The method that fermentation method prepares peony pollen polypeptide |
| CN110157758A (en) * | 2017-12-26 | 2019-08-23 | 浦江县欧立生物技术有限公司 | The purposes of the Flos Rosae Rugosas pollen polypeptide of fermentation method preparation |
| CN111588758A (en) * | 2020-07-07 | 2020-08-28 | 西藏云朵玫瑰科技有限公司 | Extraction method of rose extract |
| CN114457136A (en) * | 2022-01-27 | 2022-05-10 | 美尔健（深圳）生物科技有限公司 | Fermentation liquid based on rose fermentation recombinant collagen and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| HU 等: "Fermentation of rose residue by Lactiplantibacillus plantarum B7 and Bacillus subtilis natto promotes polyphenol content and beneficial bioactivity", JOURNAL OF BIOSCIENCE AND BIOENGINEERING, vol. 134, no. 6, pages 501 - 507 * |
| 刘婕 等: "玫瑰希腊式酸奶生产工艺研究", 中国食品添加剂, no. 02, pages 161 - 165 * |
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Application publication date: 20211119 |