CN115820783A - Preparation method for obtaining multiple triterpenoids by fermenting rosa roxburghii tratt - Google Patents
Preparation method for obtaining multiple triterpenoids by fermenting rosa roxburghii tratt Download PDFInfo
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- CN115820783A CN115820783A CN202211443227.4A CN202211443227A CN115820783A CN 115820783 A CN115820783 A CN 115820783A CN 202211443227 A CN202211443227 A CN 202211443227A CN 115820783 A CN115820783 A CN 115820783A
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- 150000003648 triterpenes Chemical class 0.000 title claims abstract description 41
- 238000002360 preparation method Methods 0.000 title claims abstract description 9
- 240000002547 Rosa roxburghii Species 0.000 title description 7
- 235000000640 Rosa roxburghii Nutrition 0.000 title description 7
- 241000220317 Rosa Species 0.000 claims abstract description 34
- 238000000855 fermentation Methods 0.000 claims abstract description 29
- 230000004151 fermentation Effects 0.000 claims abstract description 29
- 238000004821 distillation Methods 0.000 claims abstract description 6
- 238000000746 purification Methods 0.000 claims abstract description 6
- 239000002893 slag Substances 0.000 claims abstract description 4
- 239000007788 liquid Substances 0.000 claims description 21
- 230000001580 bacterial effect Effects 0.000 claims description 18
- 108090000790 Enzymes Proteins 0.000 claims description 16
- 102000004190 Enzymes Human genes 0.000 claims description 16
- CSNNHWWHGAXBCP-UHFFFAOYSA-L Magnesium sulfate Chemical compound [Mg+2].[O-][S+2]([O-])([O-])[O-] CSNNHWWHGAXBCP-UHFFFAOYSA-L 0.000 claims description 16
- 229940088598 enzyme Drugs 0.000 claims description 16
- 239000012153 distilled water Substances 0.000 claims description 14
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 9
- 229920001817 Agar Polymers 0.000 claims description 8
- 241000894006 Bacteria Species 0.000 claims description 8
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 8
- 244000017020 Ipomoea batatas Species 0.000 claims description 8
- 235000002678 Ipomoea batatas Nutrition 0.000 claims description 8
- 235000019764 Soybean Meal Nutrition 0.000 claims description 8
- 229930003270 Vitamin B Natural products 0.000 claims description 8
- 239000008272 agar Substances 0.000 claims description 8
- 229940041514 candida albicans extract Drugs 0.000 claims description 8
- 235000013399 edible fruits Nutrition 0.000 claims description 8
- 239000008103 glucose Substances 0.000 claims description 8
- 229910052943 magnesium sulfate Inorganic materials 0.000 claims description 8
- 235000019341 magnesium sulphate Nutrition 0.000 claims description 8
- 229920001592 potato starch Polymers 0.000 claims description 8
- 239000004455 soybean meal Substances 0.000 claims description 8
- 235000019156 vitamin B Nutrition 0.000 claims description 8
- 239000011720 vitamin B Substances 0.000 claims description 8
- 239000012138 yeast extract Substances 0.000 claims description 8
- 239000000706 filtrate Substances 0.000 claims description 6
- 239000001963 growth medium Substances 0.000 claims description 6
- 238000002156 mixing Methods 0.000 claims description 6
- 239000004310 lactic acid Substances 0.000 claims description 5
- 235000014655 lactic acid Nutrition 0.000 claims description 5
- 101710130006 Beta-glucanase Proteins 0.000 claims description 4
- 108010059892 Cellulase Proteins 0.000 claims description 4
- 108091005804 Peptidases Proteins 0.000 claims description 4
- 108010059820 Polygalacturonase Proteins 0.000 claims description 4
- 239000004365 Protease Substances 0.000 claims description 4
- 102100037486 Reverse transcriptase/ribonuclease H Human genes 0.000 claims description 4
- 229940106157 cellulase Drugs 0.000 claims description 4
- 108010093305 exopolygalacturonase Proteins 0.000 claims description 4
- 230000003321 amplification Effects 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 3
- 239000012528 membrane Substances 0.000 claims description 3
- 238000001471 micro-filtration Methods 0.000 claims description 3
- 238000010899 nucleation Methods 0.000 claims description 3
- 238000003199 nucleic acid amplification method Methods 0.000 claims description 3
- 239000000843 powder Substances 0.000 claims description 3
- 239000002244 precipitate Substances 0.000 claims description 3
- 238000010992 reflux Methods 0.000 claims description 3
- 230000001954 sterilising effect Effects 0.000 claims description 3
- 238000003756 stirring Methods 0.000 claims description 3
- 241000186660 Lactobacillus Species 0.000 claims description 2
- 229940039696 lactobacillus Drugs 0.000 claims description 2
- 239000011148 porous material Substances 0.000 claims description 2
- 238000000605 extraction Methods 0.000 abstract description 5
- 241000196324 Embryophyta Species 0.000 abstract description 3
- 230000000694 effects Effects 0.000 description 5
- 235000015097 nutrients Nutrition 0.000 description 5
- 150000001875 compounds Chemical class 0.000 description 4
- 244000005700 microbiome Species 0.000 description 3
- XBZYWSMVVKYHQN-MYPRUECHSA-N (4as,6as,6br,8ar,9r,10s,12ar,12br,14bs)-10-hydroxy-2,2,6a,6b,9,12a-hexamethyl-9-[(sulfooxy)methyl]-1,2,3,4,4a,5,6,6a,6b,7,8,8a,9,10,11,12,12a,12b,13,14b-icosahydropicene-4a-carboxylic acid Chemical compound C1C[C@H](O)[C@@](C)(COS(O)(=O)=O)[C@@H]2CC[C@@]3(C)[C@]4(C)CC[C@@]5(C(O)=O)CCC(C)(C)C[C@H]5C4=CC[C@@H]3[C@]21C XBZYWSMVVKYHQN-MYPRUECHSA-N 0.000 description 2
- 206010028980 Neoplasm Diseases 0.000 description 2
- 230000009286 beneficial effect Effects 0.000 description 2
- 201000011510 cancer Diseases 0.000 description 2
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 2
- 229910000396 dipotassium phosphate Inorganic materials 0.000 description 2
- 235000019797 dipotassium phosphate Nutrition 0.000 description 2
- 239000003814 drug Substances 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 208000022309 Alcoholic Liver disease Diseases 0.000 description 1
- 229920002261 Corn starch Polymers 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 235000004347 Perilla Nutrition 0.000 description 1
- 244000124853 Perilla frutescens Species 0.000 description 1
- 230000006907 apoptotic process Effects 0.000 description 1
- 235000009508 confectionery Nutrition 0.000 description 1
- 239000008120 corn starch Substances 0.000 description 1
- 230000029087 digestion Effects 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 239000003205 fragrance Substances 0.000 description 1
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 1
- 235000013402 health food Nutrition 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 239000000203 mixture Substances 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 238000010791 quenching Methods 0.000 description 1
- 230000000171 quenching effect Effects 0.000 description 1
- 210000000952 spleen Anatomy 0.000 description 1
- 238000005728 strengthening Methods 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 230000035922 thirst Effects 0.000 description 1
- 235000013619 trace mineral Nutrition 0.000 description 1
- 239000011573 trace mineral Substances 0.000 description 1
- 238000012795 verification Methods 0.000 description 1
- 239000002699 waste material Substances 0.000 description 1
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02E—REDUCTION OF GREENHOUSE GAS [GHG] EMISSIONS, RELATED TO ENERGY GENERATION, TRANSMISSION OR DISTRIBUTION
- Y02E50/00—Technologies for the production of fuel of non-fossil origin
- Y02E50/10—Biofuels, e.g. bio-diesel
Abstract
The invention discloses a preparation method for obtaining multiple triterpenoids through roxburgh rose fermentation, which belongs to the technical field of plant extraction, and comprises the steps of slag preparation, strain propagation, fermentation, purification and distillation.
Description
Technical Field
The invention relates to the technical field of plant extraction, in particular to a preparation method for obtaining multiple triterpenoids by fermenting roxburgh rose.
Background
The roxburgh rose is a fruit of a rosaceous plant, namely reeling silk, the pulp of the roxburgh rose is crisp and sweet, has thick fragrance and rich nutrition, is a good fruit for nourishing and building bodies, is mature in autumn, is cleaned after thorn removal, is removed from seeds, can be eaten fresh or dried in the sun for medicine use and the like, and has the effects of promoting digestion, strengthening the spleen, astringing to arrest diarrhea, quenching thirst, relieving summer heat, dispelling the effects of alcohol and the like. The rosa roxburghii contains various trace elements, wherein the rosa roxburghii contains triterpenes, and the triterpenes in the rosa roxburghii have the effects of inducing cancer cell apoptosis and converting cancer cells into normal cells through clinical mechanism verification.
Through retrieval, in the prior art, the Chinese patent application number: CN201710929795.8 discloses the application of multiple triterpene extracts in the preparation of drugs, health products and health foods for preventing and treating alcoholic liver diseases, but the following problems still exist in the invention:
after the fresh roxburgh rose fruit is crushed and juiced, the roxburgh rose juice has medicinal value and is used for extracting various triterpenoids too much, the content of the triterpenoids extracted from roxburgh rose pomace is low, and then the waste of various triterpenoid components in the roxburgh rose pomace is easily caused by the technical problem of extraction.
Disclosure of Invention
The invention aims to overcome the difficulties in the background technology and provide a method for producing and preparing various triterpenes, which can improve the content of various triterpenes in roxburgh rose pomace.
In order to achieve the purpose, the technical scheme is as follows: a method for preparing multiple triterpenes by fermenting fructus Rosae Normalis comprises the following steps: (1) slag preparation: preparing roxburgh rose pomace from fresh roxburgh rose fruits, and refrigerating for later use;
(2) Preparing a culture medium, namely fully mixing 200ml of distilled water and 10g of lactic acid bacteria concentrated solution which is cooled after being boiled for 5min and a plurality of complex enzymes, standing for 2-3h to obtain bacterial liquid 1, adding 5.0g of glucose, 5g of soybean meal, 5g of sweet potato starch, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO4, 0.2g of MgSO4 and 1L of distilled water into the bacterial liquid 1, fully mixing, standing for 4-6 h under the environment of pH of 5-6 and 35-37 ℃, and finally sterilizing for 30min under 0.1 Mpa to obtain bacterial liquid 2;
(3) Expanding and culturing strains: sequentially placing the bacterial liquid 2 obtained in the step (2) into a culture bottle, a seeding tank and a fermentation tank for amplification culture to finally obtain culture bacterial liquid;
(4) Fermentation: uniformly stirring the culture bacterial liquid obtained in the step (3) and 50g of roxburgh rose pomace, and naturally fermenting for 48 hours in a constant-temperature 35-37 ℃ sterile environment to obtain roxburgh rose fermentation liquid;
(5) And (3) purification: refluxing the roxburgh rose fermentation liquor obtained in the step (4) with 75% of ethanol for 2-3 times, and recovering the ethanol to obtain filtrate A;
(6) And (3) distillation: then, the filtrate A was filtered, and the precipitate was washed with distilled water 1 to 3 times and dried under reduced pressure at 40 ℃ to obtain various triterpene powders.
Further, the pomace prepared in the step (1) is 100-150 meshes.
Further, the complex enzyme in the step (2) comprises: 3g of beta-glucanase, 5g of cellulase, 5g of pectinase and 0.5g of protease.
Further, the operation environment in the step (2) is a sterile environment.
Further, in the step (3), during strain expansion culture, the multiple triterpene expansion culture medium obtained in the step (2) is cooled to room temperature.
Further, the filtration in the step (6) uses a microfiltration membrane with the pore size of 0.5-1 μm.
Further, the lactic acid bacteria solution added in the step (2) is acetic acid bacteria concentrated solution.
The beneficial effect who adopts above-mentioned scheme does: the preparation method for obtaining multiple triterpenoids through fermentation of the rosa roxburghii comprises the steps of preparing residues, expanding culture of strains, fermenting, purifying and distilling, multiple complex enzymes, lactic acid bacteria and multiple nutritional ingredients are added in the step of preparing the culture medium, the effect of expanding culture and pre-fermenting the living environment of the strains in the subsequent fermentation step is achieved, the propagation speed of microorganisms is increased, and finally multiple triterpenoids in the rosa roxburghii fruit residues are better extracted.
Detailed Description
The technical solution of the present invention will be clearly and completely described below with reference to the specific embodiments of the present invention. The described embodiments are only some, not all embodiments of the invention. 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.
Example 1
A method for preparing multiple triterpenes by fermenting fructus Rosae Normalis comprises the following steps:
(1) Preparing slag: preparing roxburgh rose pomace from fresh roxburgh rose fruits, and refrigerating for later use;
(2) Preparing a culture medium, namely fully mixing 200ml of distilled water and 10g of lactobacillus concentrated solution which is boiled for 5min and cooled and a plurality of complex enzymes, standing for 2-3h to obtain a bacterial liquid 1, adding 5.0g of glucose, 5g of soybean meal, 5g of sweet potato starch, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO, 0.2g of MgSO4 and 1L of distilled water into the bacterial liquid 1, fully mixing, standing for 4-6 h under the environment of pH 5-6 and 35-37 ℃, and finally sterilizing for 30min under 0.1 Mpa to obtain a bacterial liquid 2;
(3) Expanding and culturing strains: sequentially placing the bacterial liquid 2 obtained in the step (2) into a culture bottle, a seeding tank and a fermentation tank for amplification culture to finally obtain culture bacterial liquid;
(4) And (3) fermentation: uniformly stirring the culture bacterial liquid obtained in the step (3) and 50g of roxburgh rose pomace, and naturally fermenting for 48 hours in a constant-temperature 35-37 ℃ sterile environment to obtain roxburgh rose fermentation liquid;
(5) And (3) purification: refluxing the roxburgh rose fermentation liquor obtained in the step (4) with 75% of ethanol for 2-3 times, and recovering the ethanol to obtain filtrate A;
(6) And (3) distillation: then, the filtrate A was filtered, and the precipitate was washed with distilled water 1 to 3 times and dried under reduced pressure at 40 ℃ to obtain various triterpene powders.
In the invention, the following control group experiments are made, wherein (1) the roxburgh rose pomace is directly extracted with various triterpenoids, (2) the roxburgh rose pomace is subjected to common fermentation to extract various triterpenoids, (3) the roxburgh rose pomace is added with various complex enzymes in the step (2) to extract various triterpenoids (the complex enzymes comprise beta-glucanase 3g, cellulase 5g, pectinase 5g and protease 0.5 g.), 50g of roxburgh rose pomace is subjected to still standing fermentation for 3 hours at 37 ℃ under the same environment of natural pH, then purification and distillation are carried out, three groups of control groups are subjected to extraction of various triterpenoids and content recording, and the following experimental data are obtained:
| content/item | Not fermenting | General fermentation | Complex enzyme fermentation |
| Content of various triterpenes | 6.13% | 8.67% | 13.28% |
From the experimental results, the content of various triterpenoids extracted from the roxburgh rose pomace with the same gram weight is obviously increased, and the content of various triterpenoids in the compound enzyme fermentation is greater than that in the common fermentation and is greater than that in the non-fermentation state, so that the compound enzyme can actually improve the content of various triterpenoids when the roxburgh rose pomace is extracted.
Because fruits contain less nutrient substances and the propagation rate of microorganisms is low, after various complex enzymes are added to the pomace, the following control groups are made to perform experimental selection on the nutrient materials added in the step (2) in an expanding culture mode: (in the experimental step, 50g of roxburgh rose pomace is selected to mix roxburgh rose pomace, the same compound enzyme → different nutrient components from (1) - (5) are added → the mixture is naturally fermented for 48h under the aseptic environment with the constant temperature of 35-37 ℃ to obtain roxburgh rose fermentation liquor → the content of various triterpenoids is recorded by purification and distillation)
(1) 5.0g of glucose, 5g of soybean meal, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO4, 0.2g of MgSO4 and 1L of distilled water
(2) 5.0g of glucose, 5g of soybean meal, 5g of sweet potato starch, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO, 0.2g of MgSO4 and 1L of distilled water
(3) 5.0g of glucose, 5g of soybean meal, 5g of corn starch, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO, 0.2g of MgSO4 and 1L of distilled water
(4) 5.0g of glucose, 5g of soybean meal, 5g of perilla leaf, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO, 0.2g of MgSO4 and 1L of distilled water
(5) No addition of nutrients
The experimental data are as follows:
| content/item | Control group (1) | Control group (2) | Control group (3) | Control group (4) | Control group (5) |
| Content of various triterpenes | 14.24% | 18.27% | 15.28% | 16.42% | 12.36% |
According to the experimental result, in the component selection, the content of various triterpenoids extracted by fermentation in the subsequent fermentation after the sweet potato starch is used is the highest, and is far greater than that in the fermentation process after only the compound enzyme is used, so that the fermentation process in the subsequent step (3) is more suitable after the sweet potato starch is added, and the nutrient components in the sweet potato starch are more beneficial to the fermentation of the triterpenoid microorganisms.
Therefore, the two experiments show that the complex enzyme, 5.0g of glucose, 5g of soybean meal, 5g of sweet potato starch, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO, 0.2g of MgSO4 and 1L of distilled water added in the step (2) have the effect of improving the extraction content of various triterpenoids.
Example 2
The pomace prepared in the step (1) is 100-150 meshes.
The complex enzyme in the step (2) comprises the following steps: 3g of beta-glucanase, 5g of cellulase, 5g of pectinase and 0.5g of protease.
The operation environment in the step (2) is a sterile environment.
In the step (3), during strain propagation, the multiple triterpene propagation mediums obtained in the step (2) are cooled to room temperature.
The microfiltration membrane with the aperture of 0.5-1 mu m is used for filtering in the step (6).
The lactic acid bacteria solution added in the step (2) is acetic acid bacteria concentrated solution.
It will be evident to those skilled in the art that the invention is not limited to the details of the foregoing illustrative embodiments, and that the present invention may be embodied in other specific forms without departing from the spirit or essential attributes thereof. The present embodiments are therefore to be considered in all respects as illustrative and not restrictive, the scope of the invention being indicated by the appended claims rather than by the foregoing description, and all changes which come within the meaning and range of equivalency of the claims are therefore intended to be embraced therein.
Furthermore, it should be understood that although the present description refers to embodiments, not every embodiment may contain only a single embodiment, and such description is for clarity only, and those skilled in the art should integrate the description, and the embodiments may be combined as appropriate to form other embodiments understood by those skilled in the art.
Claims (7)
1. A preparation method for obtaining a plurality of triterpenoids by fermenting roxburgh rose is characterized by comprising the following steps: the method comprises the following steps:
(1) Preparing slag: preparing roxburgh rose pomace from fresh roxburgh rose fruits, and refrigerating for later use;
(2) Preparing a culture medium, namely fully mixing 200ml of distilled water and 10g of lactobacillus concentrated solution which is boiled for 5min and cooled and a plurality of complex enzymes, standing for 2-3h to obtain a bacterial liquid 1, adding 5.0g of glucose, 5g of soybean meal, 5g of sweet potato starch, 5g of agar, 0.01g of yeast extract, 0.01g of vitamin B, 1.5g of K2HPO, 0.2g of MgSO4 and 1L of distilled water into the bacterial liquid 1, fully mixing, standing for 4-6 h under the environment of pH 5-6 and 35-37 ℃, and finally sterilizing for 30min under 0.1 Mpa to obtain a bacterial liquid 2;
(3) Expanding and culturing strains: sequentially placing the bacterial liquid 2 obtained in the step (2) into a culture bottle, a seeding tank and a fermentation tank for amplification culture to finally obtain culture bacterial liquid;
(4) Fermentation: uniformly stirring the culture bacterial liquid obtained in the step (3) and 50g of roxburgh rose pomace, and naturally fermenting for 48 hours in a constant-temperature 35-37 ℃ sterile environment to obtain roxburgh rose fermentation liquid;
(5) And (3) purification: refluxing the roxburgh rose fermentation liquor obtained in the step (4) with 75% of ethanol for 2-3 times, and recovering the ethanol to obtain filtrate A;
(6) And (3) distillation: then, the filtrate A was filtered, and the precipitate was washed with distilled water 1 to 3 times and dried under reduced pressure at 40 ℃ to obtain various triterpene powders.
2. The method of producing a plurality of triterpenes according to claim 1, wherein: the pomace prepared in the step (1) is 100-150 meshes.
3. The method of producing a plurality of triterpenes according to claim 1, wherein: the complex enzyme in the step (2) comprises the following steps: 3g of beta-glucanase, 5g of cellulase, 5g of pectinase and 0.5g of protease.
4. The method of producing a plurality of triterpenes according to claim 1, wherein: the operation environment in the step (2) is aseptic environment.
5. The method of producing a plurality of triterpenes according to claim 1, wherein: in the step (3), the strain expansion culture needs to be carried out, and the multiple triterpene expansion culture medium obtained in the step (2) is cooled to room temperature.
6. The method of producing a plurality of triterpenes according to claim 4, wherein: the microfiltration membrane with the pore size of 0.5-1 mu m is used for filtering in the step (6).
7. The method of producing a plurality of triterpenes according to claim 1, wherein: the lactic acid bacteria solution added in the step (2) is acetic acid bacteria concentrated solution.
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Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
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| CN117683651A (en) * | 2024-01-26 | 2024-03-12 | 东北林业大学 | Genetically engineered bacterium and construction method and application thereof |
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| Title |
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| CN117683651A (en) * | 2024-01-26 | 2024-03-12 | 东北林业大学 | Genetically engineered bacterium and construction method and application thereof |
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Inventor after: Lin Youjun Inventor after: Chen Huiru Inventor before: Lin Youjun Inventor before: Chen Huiru |