CN111893153A - Method for extracting ademetionine fermentation liquor by breaking cell wall - Google Patents
Method for extracting ademetionine fermentation liquor by breaking cell wall Download PDFInfo
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- CN111893153A CN111893153A CN202010819067.3A CN202010819067A CN111893153A CN 111893153 A CN111893153 A CN 111893153A CN 202010819067 A CN202010819067 A CN 202010819067A CN 111893153 A CN111893153 A CN 111893153A
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- fermentation liquor
- ademetionine
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- extraction
- breaking
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- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
- C12P19/00—Preparation of compounds containing saccharide radicals
- C12P19/26—Preparation of nitrogen-containing carbohydrates
- C12P19/28—N-glycosides
- C12P19/38—Nucleosides
- C12P19/40—Nucleosides having a condensed ring system containing a six-membered ring having two nitrogen atoms in the same ring, e.g. purine nucleosides
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H1/00—Processes for the preparation of sugar derivatives
- C07H1/06—Separation; Purification
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- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07H—SUGARS; DERIVATIVES THEREOF; NUCLEOSIDES; NUCLEOTIDES; NUCLEIC ACIDS
- C07H19/00—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof
- C07H19/02—Compounds containing a hetero ring sharing one ring hetero atom with a saccharide radical; Nucleosides; Mononucleotides; Anhydro-derivatives thereof sharing nitrogen
- C07H19/04—Heterocyclic radicals containing only nitrogen atoms as ring hetero atom
- C07H19/16—Purine radicals
- C07H19/167—Purine radicals with ribosyl as the saccharide radical
Abstract
The invention discloses a method for extracting adenosine methionine fermentation liquor by breaking cell walls. The method for extracting the adenosine methionine fermentation liquor by breaking the wall comprises the steps of culturing saccharomyces cerevisiae in a fermentation tank for 30-80 hours, supplementing glucose and L-methionine during the culture period, and directly using the fermentation liquor for post-extraction; introducing yeast fermentation liquor into the ceramic membrane; after concentration; the concentration of the bacteria is 30-60%; adding water into the high-bacteria-concentration feed liquid, continuously concentrating until the dialysate is colorless and transparent, adding an organic solvent into the ceramic membrane, circulating for 20-50min, and continuously concentrating to obtain a clear liquid to obtain an extract; wherein the ratio of the organic solvent to the solid content is 1: 1.3-1.8. The method has the advantages of simple steps, low requirement on equipment, great saving of production cost such as manpower, material resources and production period, high extraction yield and effective improvement of product profit.
Description
Technical Field
The invention relates to the technical field of biological pharmacy, in particular to a method for extracting adenosine methionine fermentation liquor by breaking walls.
Background
S-Adenosyl-L-methionine (SAM) was discovered in 1953 as a sulfonyl compound in the form of an organic quaternary sulfur of methionine. It is widely present in various organisms and is an important physiologically active substance in the human body. Biologically, SAM is a one-, three-and five-carbon group donor, with methyl-, propyl-, and sulfur-transfer physiological effects.
Its chemical name is "+/-5 '- [ (R) -3-amino-3-carboxypropyl ] methylsulfonyl ] -5' -deoxyadenosine 1, 4-butanedisulfonate"
English name ± -5'- [ (R) -, 3-amino-3-carboxypropyl ] methysulfonio ] -5' -deoxysylnine inner salt,1, 4-butandisultaine
Chemical structure
At present, the domestic demand for 1, 4-butanedisulfonic acid adenosine methionine is large, but the productivity is insufficient, and under the condition based on the prior art, the extraction mode with high yield and simple operation on the adenosine methionine fermentation liquor needs to be researched and invented.
Disclosure of Invention
According to one aspect of the invention, the method for extracting the ademetionine fermentation liquor by breaking the wall comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 30-80h, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing the yeast fermentation liquor into the ceramic membrane; after concentration; the concentration of the bacteria is 30-60%;
s3, adding water into the high-bacteria-concentration feed liquid, continuously concentrating until the dialysate is colorless and transparent, adding the organic solvent into the ceramic membrane, circulating for 20-50min, and continuously concentrating to obtain clear liquid to obtain an extract; wherein the ratio of the organic solvent to the solid content is 1: 1.3-1.8.
In some embodiments, the Saccharomyces cerevisiae in step S1 is Saccharomyces cerevisiae (ATCC 11909).
In some embodiments, the ceramic membrane in step S2 has a pore size of 8-100 nm.
In some embodiments, the colorless standard in step S3 is less than or equal to standard colorimetric solution Y-3.
In some embodiments, the solvent in step S3 is an ester solvent or a ketone solvent.
In some embodiments, the solid content in step S3 is the product of the total volume of the fermentation broth and the microbial concentration of the fermentation broth.
In some embodiments, in step S3, the high-bacteria-concentration feed liquid is further concentrated until the dialysate is colorless and transparent, the organic solvent is added into the ceramic membrane, the circulation is performed for 20-25min, and the clear liquid is further concentrated to obtain an extract; wherein the ratio of the organic solvent to the solid content is 1: 1.3-1.5.
In some embodiments, the ester solvent and ketone solvent in step S3 are ethyl acetate, acetone or butanone.
The method for extracting the adenosine methionine fermentation liquor by breaking the wall has the advantages that the fermentation liquor of the adenosine methionine producing bacteria is subjected to ceramic membrane filtration to obtain high-bacteria-concentration fermentation liquor; adding organic solvent solution, circulating, filtering and collecting filtrate. The method has simple steps, low requirements on equipment, high extraction yield and the like, greatly saves the production cost of manpower, material resources, production period and the like, and effectively improves the product profit.
Detailed Description
The present invention will be described in further detail with reference to examples.
Example 1 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 30 hours, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 20% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 10 nm; after concentration; the concentration of the bacteria is 30 percent;
s3, adding water to the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y2, adding ethyl acetate into the high-bacteria-concentration feed liquid according to a ratio of 1:1.3 and 30.7L, and circulating for 20 min; continuously concentrating to obtain clear liquid to obtain extract;
s4, the extraction rate is 83%.
Example 2 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 50 hours, supplementing glucose and L-methionine during the culturing, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 18% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 50 nm; after concentration; the concentration of the bacteria is 60%;
s3, adding water to the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y3, adding butyl acetate into the high-bacteria-concentration feed liquid according to a ratio of 1:1.8 and 20L, circulating for 50min, and continuously concentrating to obtain clear liquid to obtain extract;
s4, the extraction rate is 80%.
Example 3 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 40 hours, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 20% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 50 nm; after concentration; the concentration of the bacteria is 45%;
s3, adding water into the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y2, adding acetone into the high-bacteria-concentration feed liquid at a ratio of 1:1.8 and 22.2L, circulating for 40min, and continuously concentrating to obtain clear liquid to obtain extract;
(4) the extraction rate was 85%.
Example 4 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 66 hours, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 24% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 100 nm; after concentration; the concentration of the bacteria is 57%;
s3, supplementing water to the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y3, adding butanone into the high-bacteria-concentration feed liquid according to a ratio of 1:1.3 and 36.9L, circulating for 50min, and continuously concentrating to obtain clear liquid to obtain an extracting solution;
s4, the extraction rate is 87%.
Example 5 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 80 hours, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 15% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 50 nm; after concentration; the concentration of the bacteria is 61%;
s3, adding water into the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y2, adding butyl acetate into the high-bacteria-concentration feed liquid according to a ratio of 1:1.8 and 16.7L, circulating for 30min, and continuously concentrating to obtain clear liquid to obtain an extracting solution;
s4, the extraction rate is 81%.
Example 6 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 30 hours, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 22% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 10 nm; after concentration; the concentration of the bacteria is 35 percent;
s3, adding water into the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y3, adding acetone into the high-bacteria-concentration feed liquid according to a ratio of 1:1.3 and total 33.8L, circulating for 35min, and continuously concentrating to obtain clear liquid to obtain extract;
s4, the extraction rate is more than 83%.
Example 7 method for extracting ademetionine fermentation broth by breaking cell wall
The method comprises the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 30 hours, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing 200L of 22% bacteria-concentrated ademetionine fermentation liquor into a ceramic membrane with the aperture of 10 nm; after concentration; the concentration of the bacteria is 35 percent;
s3, adding water into the high-bacteria-concentration feed liquid, continuously concentrating to obtain dialysate Y3, adding acetone into the high-bacteria-concentration feed liquid according to a ratio of 1:1.3 and total 33.8L, circulating for 35min, and continuously concentrating to obtain clear liquid to obtain extract;
s4, the extraction rate is more than 83%.
According to the method for extracting the adenosine methionine fermentation liquor by breaking the wall, the method for extracting the adenosine methionine fermentation liquor by breaking the wall comprises the steps of culturing saccharomyces cerevisiae in a fermentation tank for 30-80 hours, supplementing glucose and L-methionine during the culture period, and directly using the fermentation liquor for post-extraction; introducing yeast fermentation liquor into the ceramic membrane; after concentration; the concentration of the bacteria is 30-60%; adding water into the high-bacteria-concentration feed liquid, continuously concentrating until the dialysate is colorless and transparent, adding an organic solvent into the ceramic membrane, circulating for 20-50min, and continuously concentrating to obtain a clear liquid to obtain an extract; wherein the ratio of the organic solvent to the solid content is 1: 1.3-1.8. The method has simple steps, low requirements on equipment, high extraction yield and the like, greatly saves the production cost of manpower, material resources, production period and the like, and effectively improves the product profit.
What has been described are merely some of the embodiments of the present invention.
It will be apparent to those skilled in the art that various modifications and improvements can be made without departing from the inventive concept of the present invention, and these modifications and improvements are intended to be within the scope of the invention.
Claims (8)
1. A method for extracting ademetionine fermentation liquor by breaking cell walls is characterized by comprising the following steps:
s1, culturing the saccharomyces cerevisiae in a fermentation tank for 30-80h, supplementing glucose and L-methionine during the culture, and directly using the fermentation liquor for post-extraction;
s2, introducing the yeast fermentation liquor into the ceramic membrane; after concentration; the concentration of the bacteria is 30-60%;
s3, adding water into the high-bacteria-concentration feed liquid, continuously concentrating until the dialysate is colorless and transparent, adding the organic solvent into the ceramic membrane, circulating for 20-50min, and continuously concentrating to obtain clear liquid to obtain an extract; wherein the ratio of the organic solvent to the solid content is 1: 1.3-1.8.
2. The method for extracting ademetionine fermentation broth by wall breaking according to claim 1, wherein the Saccharomyces cerevisiae (ATCC11909) is Saccharomyces cerevisiae in step S1.
3. The method for wall-breaking extraction of ademetionine fermentation broth according to claim 2, wherein the pore size of the ceramic membrane in step S2 is 8-100 nm.
4. The method for wall-broken extraction of ademetionine fermentation broth according to claim 1, wherein the standard of colorless in step S3 is below standard colorimetric solution Y-3.
5. The method for breaking the wall of the adenosyl methionine fermentation broth for extraction according to claim 1, wherein the solvent in step S3 is an ester solvent or a ketone solvent.
6. The method for wall-broken extraction of ademetionine fermentation liquor according to claim 1, wherein the solid content in step S3 is the product of the total volume of the fermentation liquor and the microbial concentration of the fermentation liquor.
7. The method for wall-breaking extraction of ademetionine fermentation broth according to claim 1, wherein in step S3, the high-concentration feed liquid is supplemented with water and continuously concentrated until the dialysate is colorless and transparent, the organic solvent is added into the ceramic membrane, the circulation is carried out for 20-25min, and the supernatant is continuously concentrated to obtain an extract; wherein the ratio of the organic solvent to the solid content is 1: 1.3-1.5.
8. The method for wall-breaking extraction of ademetionine fermentation broth according to claim 5, wherein said ester solvent and ketone solvent are ethyl acetate, acetone or butanone.
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Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102747123A (en) * | 2012-07-31 | 2012-10-24 | 无锡福祈制药有限公司 | Process for preparing ademetionine butanedisulfonate |
| CN104131052A (en) * | 2014-08-06 | 2014-11-05 | 西藏金稞集团有限责任公司 | Method for producing ademetionine by fermentation of saccharomycetes |
| CN106349311A (en) * | 2016-08-23 | 2017-01-25 | 北京金阳利康医药有限公司 | Method for extracting S-adenosylmethionine from yeast fermentation liquor |
-
2020
- 2020-08-14 CN CN202010819067.3A patent/CN111893153A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102747123A (en) * | 2012-07-31 | 2012-10-24 | 无锡福祈制药有限公司 | Process for preparing ademetionine butanedisulfonate |
| CN104131052A (en) * | 2014-08-06 | 2014-11-05 | 西藏金稞集团有限责任公司 | Method for producing ademetionine by fermentation of saccharomycetes |
| CN106349311A (en) * | 2016-08-23 | 2017-01-25 | 北京金阳利康医药有限公司 | Method for extracting S-adenosylmethionine from yeast fermentation liquor |
Non-Patent Citations (2)
| Title |
|---|
| 刘勇等: "高产 S - 腺苷甲硫氨酸酵母菌株的筛选", 《食品与发酵工业》 * |
| 林海军: "酿酒酵母发酵生产腺苷蛋氨酸及其分离纯化工艺的研究", 《中国优秀硕士学位论文全文数据库 工程科技I辑》 * |
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| CB02 | Change of applicant information |
Address after: 219 Furong Zhongsi Road, Xishan Economic and Technological Development Zone, Wuxi City, Jiangsu Province, 214000 Applicant after: Zhuohe Pharmaceutical Group Co.,Ltd. Address before: 219 Furong Zhongsi Road, Xishan Economic and Technological Development Zone, Wuxi City, Jiangsu Province, 214000 Applicant before: Zhuohe Pharmaceutical Group Co.,Ltd. |
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Application publication date: 20201106 |
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