CN103614308A - Saccharomyces cerevisiae for ATP synthesis and applications thereof - Google Patents
Saccharomyces cerevisiae for ATP synthesis and applications thereof Download PDFInfo
- Publication number
- CN103614308A CN103614308A CN201310357563.1A CN201310357563A CN103614308A CN 103614308 A CN103614308 A CN 103614308A CN 201310357563 A CN201310357563 A CN 201310357563A CN 103614308 A CN103614308 A CN 103614308A
- Authority
- CN
- China
- Prior art keywords
- atp
- cervisiae
- saccharomyces cerevisiae
- synthetic
- applications
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Granted
Links
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
Abstract
The invention discloses a kind of Saccharomyces cerevisiae for ATP synthesis and applications thereof. The Saccharomyces cerevisiae can synthesize ATP efficiently, solves the problem of high cost of ATP synthesis, and has wide industrial application values.
Description
Technical field
The invention belongs to microorganism field, in particular to a kind of for the synthetic S. cervisiae of ATP and application thereof.
Background technology
ATP synthetic method is mainly divided into chemical synthesis, enzyme catalysis method and microorganism fermentation system:
Chemical synthesis
Be generally compound with phosphate group as phosphate to body, under the catalysis of catalyzer, phosphoric acid is transferred on reaction substrate AMP or ADP after by selectivity activatable and is generated ATP to the phosphate group of body, reaction specificity is strong, product is easily separated, but phosphorylation agent box catalyzer is more expensive, transformation efficiency is relatively low, and its lytic activity less stable, so hinder the application of the method in ATP suitability for industrialized production.
Biological synthesis process
Enzyme catalysis method
The synthetic ATP elementary process of enzyme catalysis be phosphate donor under the catalysis of phosphorylating kinase, phosphate is transferred to AMP or ADP, generate ATP, the method is one of focus in ATP study on the synthesis, at present, expensive due to enzyme, is not used widely industrial yet.
Photophosphorylation synthesis method
The method is to provide under the condition of illumination, utilizes the chloroplast(id) of chromatophore separated in photosynthetic bacterium or separation from plant tissue, and the alga cells with light autophyting ability, by substrate A MP or ADP and inorganic phosphate reaction generation ATP.The features such as it is abundant that the method has raw material sources, with low cost are also one of focuses of research at present.
Oxidative phosphorylation synthesis method
The method is to utilize eukaryotic plastosome, under the energy providing in respiration by substrate A MP or ADP phosphoric acid and the synthetic ATP of inorganic phosphate reaction.But due to eukaryotic plastosome separation difficulty, and the difficult problem such as poor stability limits its application in industrialization.
Utilize microbial enzyme system fermentation synthesis method
The features such as the method is directly to utilize microorganism cells as enzyme source Synthesis ATP, different from single enzyme catalysis, has the cost of conversion low, and changing effect is good, in industry, application is more wide.
1, Brevibacterium ammoniagenes
The people such as Qiao Binfu utilize Brevibacterium ammoniagenes to add the synthetic ATP of VITAMIN B4 in substratum, and in fermented liquid, ATP concentration reaches 2g/L (3.94mmol/L), and the problem of existence is that concentration of substrate is low, and product is not easily separated, and total recovery is low etc.
2, bread yeast
The people such as Liao Xianyan, in the research of the influence factor of the synthetic ATP of bread yeast, be take AMP as the synthetic ATP of substrate, and production concentration is up to 9.02mmol/L.The people such as Li Cang utilize bread leaven matricyte enzyme system as enzyme source, take AMP as the synthetic ATP of substrate conversion, and reaction solution production concentration reaches 10mmol/L.
3, cereuisiae fermentum
The people such as Zhu Jiarong utilize Immobilized Saccharomyces cerevisiae for enzyme source, take VITAMIN B4 as the synthetic ATP of substrate conversion, and its object product A TP is up to 2.46g/L (4.85mmol/L)
The people such as Li Liqi utilize cerevisiae for enzyme source, take adenosine as the synthetic ATP of substrate conversion, more than product ultimate density reaches 68.1mmol/L.
How comprehensive above background technology, obtain efficient microorganism and synthesize ATP and remain those skilled in the art's technical problem urgently to be resolved hurrily.
Summary of the invention
The object of this invention is to provide the synthetic S. cervisiae of a kind of ATP of can be used in and application thereof.
S. cervisiae of the present invention is the S. cervisiae of deposit number CGMCC NO.7397, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, or there is the S. cervisiae of 95% above affinity with the S. cervisiae of CGMCC NO. 7397.
The present invention also relates to the application of above-mentioned S. cervisiae in synthetic ATP on the other hand.
S. cervisiae of the present invention can be synthesized ATP efficiently, has solved the synthetic high problem of cost of ATP, has industrial application value widely.
Microorganism information
The S. cervisiae of using in a preferred embodiment of the present invention and embodiment (Saccharomyces cerevisiae) has been preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, deposit number CGMCC NO.7397, preservation address is: No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, Institute of Microorganism, Academia Sinica, preservation date is on April 1st, 2013.
Yeast strain CGMCC NO.7397 adopts following flow process to carry out seed selection:
Dull and stereotyped the sieve again → → wheat juice agar plate separation and purification of primary election → → liquid fermenting of pedotheque → → enrichment culture → → wort agar → → product enzymic fermentation cultivation → → Conversion of Adenosine test → → pilot plant test.
Examine under a microscope, this somatic cells is spherical in shape, and acrodolichomelia is grown, and on solid medium, this bacterium bacterium colony is oyster white, surface smoothing, and neat in edge ，Jing Institute of Microorganism, Academia Sinica is accredited as yeast saccharomyces cerevisiae.Utilize this bacterial strain can take adenosine as starting raw material synthesizing adenosine triphosphoric acid.
Strains A TP synthesis capability test of the present invention
(1) condition determination detects according to high performance liquid chromatography (Chinese Pharmacopoeia 2010 editions):
1. the reverse post of chromatographic column condition: C18,250mm*4mm
2. chromatographic condition and system availability test: be weighting agent with octadecylsilane chemically bonded silica; With 0.2mol/L phosphate buffered saline buffer, (get Sodium phosphate dibasic 35.8g, potassium primary phosphate 13.6g, adds water 900mL and dissolves, with 1mol/L sodium hydroxide solution, regulate pH value to 7.0, adding Tetrabutyl amonium bromide 1.61g, add water to 1000mL, shake up)-methyl alcohol (95: 5) is for moving phase; 35 ℃ of column temperatures, flow velocity is 1mL/min, detection wavelength is 259nm, theoretical plate number is calculated and is not less than 1500 by Triphosaden peak, go out peak order and be followed successively by adenylic acid sodium, adenosine diphosphate (ADP) disodium and Sodium ATP, the resolution of each chromatographic peak should meet the requirements.
(2) measuring method
Total nucleotide: it is appropriate to get this product, accurately weighed, add 0.1mol/L phosphate buffered saline buffer and (get SODIUM PHOSPHATE, MONOBASIC 35.8g, add water to 1000mL, anhydrous potassium dihydrogenphosphate 13.6g, adds water to 1000mL, the mutual adjust pH to 7.0 of two liquid) make to dissolve and quantitatively dilute and make the solution that contains 20ug in every mL, according to ultraviolet visible spectrophotometry (appendix IVA), measure, at the wavelength place of 259nm, measure absorbancy, by the uptake factor of C10H14N5Na2O13P3
be 279 calculating;
The weight ratio of Sodium ATP: measure according to high performance liquid chromatography (appendix VD); it is appropriate to get this product; accurately weighed; add that moving phase is dissolved and quantitatively dilution make in every 1mL the solution containing 0.4mg; get 10ul injection liquid chromatography; record color atlas, be calculated as follows the weight ratio of Sodium ATP (TATP) in total nucleotide.
T
1: adenylic acid sodium peak area
T
2: adenosine diphosphate (ADP) disodium peak area
T
aTP: Sodium ATP peak area
T
n: the peak area of other materials
Be calculated as follows Sodium ATP content in sample:
Sodium ATP content (%)=total nucleotide * Sodium ATP weight ratio * 100%
3, somatic cells enzyme activity determination method
The adenosine (0.5%) of take is substrate, phosphoric acid salt (50mmol/L) is damping fluid, under 35 ℃ of water bath condition, react after 2 hours, with 3N hydrochloric acid adjust pH to 3.0 left and right, termination reaction, stop buffer is processed through ultrasonic disruption (800W, 5min), the centrifugal 30min of 10000rpm, with testing goal product content after 50 times of purified water dilutions.
The bacterial strain that utilization screens is enzyme source, take adenosine as substrate, synthesizes ATP test under the Synthesis condition of ATP, and result shows that in reaction solution, target product accumulated concentrations is up to 135.8mmol/L.
The above is the preferred embodiments of the present invention; it should be pointed out that for those skilled in the art, do not departing under the prerequisite of principle of the present invention; can also make some improvements and modifications, these improvements and modifications also should be considered as protection scope of the present invention.
Claims (2)
1. S. cervisiae, it is the S. cervisiae of deposit number CGMCC NO.7397, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, or there is the S. cervisiae of 95% above affinity with the S. cervisiae of CGMCC NO.7397.
2. the application of S. cervisiae claimed in claim 1 in synthetic ATP.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310357563.1A CN103614308B (en) | 2013-08-16 | 2013-08-16 | S. cervisiae and its application for ATP synthesis |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310357563.1A CN103614308B (en) | 2013-08-16 | 2013-08-16 | S. cervisiae and its application for ATP synthesis |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN103614308A true CN103614308A (en) | 2014-03-05 |
| CN103614308B CN103614308B (en) | 2017-07-07 |
Family
ID=50165028
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310357563.1A Active CN103614308B (en) | 2013-08-16 | 2013-08-16 | S. cervisiae and its application for ATP synthesis |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103614308B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108120833A (en) * | 2018-02-24 | 2018-06-05 | 江南大学 | A kind of method of quick detection yeast activity |
| CN108559714A (en) * | 2018-06-01 | 2018-09-21 | 江南大学 | A kind of Yeast strain of beer with high anti-oxidation ability |
Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6070095A (en) * | 1983-09-26 | 1985-04-20 | Toray Ind Inc | Production of adenosine triphosphate |
-
2013
- 2013-08-16 CN CN201310357563.1A patent/CN103614308B/en active Active
Patent Citations (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JPS6070095A (en) * | 1983-09-26 | 1985-04-20 | Toray Ind Inc | Production of adenosine triphosphate |
Non-Patent Citations (3)
| Title |
|---|
| 段学辉 等: "增强细胞渗透性对酿酒酵母ATP", 《南 昌 大 学 学报 (工 科 版 )》 * |
| 赵伟 等: "利用酿酒酵母合成三磷酸腺苷的研究", 《广东化工》 * |
| 阮莉娇: "固定化酵母合成ATP的研究", 《中国优秀硕士学位论文全文数据库》, no. 5, 15 September 2005 (2005-09-15) * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN108120833A (en) * | 2018-02-24 | 2018-06-05 | 江南大学 | A kind of method of quick detection yeast activity |
| CN108559714A (en) * | 2018-06-01 | 2018-09-21 | 江南大学 | A kind of Yeast strain of beer with high anti-oxidation ability |
| CN108559714B (en) * | 2018-06-01 | 2019-11-08 | 江南大学 | A kind of Yeast strain of beer with high anti-oxidation ability |
Also Published As
| Publication number | Publication date |
|---|---|
| CN103614308B (en) | 2017-07-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN105112303B (en) | A kind of Aspergillus niger strain of production wine complex enzyme | |
| CN103865855A (en) | Bacillus subtilis strain and applications thereof | |
| Leite et al. | Breakfast of champions: Fast lipid accumulation by cultures of Chlorella and Scenedesmus induced by xylose | |
| CN103409485A (en) | Method for improving adenosine fermentation output through feeding organic nitrogen source | |
| CN102994467A (en) | L-d-glutamic oxidase with substrate specificity and alpha-oxoglutarate produced by catalysis of same | |
| CN104342390A (en) | Sinorhizobium meliloti strain and composition and application of sinorhizobium meliloti strain | |
| CN101294143B (en) | Bacterial strain for preparing 3-hydroxy butanone and uses thereof | |
| CN102533612B (en) | Clostridium beijerinckii strain and screening method and use thereof | |
| CN103146786A (en) | Method for producing adenosine by sequentially controlling fermentation with gradient pH | |
| Noparat et al. | Potential for using enriched cultures and thermotolerant bacterial isolates for production of biohydrogen from oil palm sap and microbial community analysis | |
| CN102719378B (en) | Method for preparing (-) gamma-lactam by catalysis asymmetry of microorganism | |
| CN103898013B (en) | The preparation of bacterium and kappa-carrageenan enzyme is revolved in one strain sea | |
| CN103614308A (en) | Saccharomyces cerevisiae for ATP synthesis and applications thereof | |
| CN110055195A (en) | The microbial strains BF-1801 of one high-efficiency degradation cellulose | |
| CN109797121A (en) | The microbial strains LM-1801 of one plant of degraded cellulose and its application | |
| CN104911135A (en) | Trehalose synthase production strain and application thereof | |
| CN104293867A (en) | Microbial synthesis method of 2'-deoxy-5'-floxuridine | |
| CN103409486A (en) | Method for improving adenosine fermentation output through feeding hypoxanthine | |
| CN101768587A (en) | Method for screening mutant strain of butanol producing strain for removing catabolic repression of glucose | |
| CN103525727A (en) | Method for transforming 1,3-propylene glycol into 3-hydroxypropionic acid by applying resting cells | |
| CN103468606B (en) | Klebsiella oxytoca and application thereof in allitol production | |
| CN103820506A (en) | Method for producing coenzyme Q10 by fermenting genetic recombinant bacteria | |
| CN103993055A (en) | Biosynthesis method of ademetionine | |
| CN105779363B (en) | A kind of method of liquefied Serratia and its Synthesis heliotropin | |
| CN112280696B (en) | Culture method for improving oil content of saccharomyces cerevisiae by maintaining ultralow-concentration organic carbon source and application of culture method |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant | ||
| CP01 | Change in the name or title of a patent holder |
Address after: 453003 No. 23, Chemical Road, Henan, Xinxiang Patentee after: Xinxiang Tuo Xin Pharmaceutical Limited by Share Ltd Address before: 453003 No. 23, Chemical Road, Henan, Xinxiang Patentee before: Xinxiang Tuoxin Biochemical Co., Ltd. |
|
| CP01 | Change in the name or title of a patent holder |