CN104004700A - High-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof - Google Patents
High-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof Download PDFInfo
- Publication number
- CN104004700A CN104004700A CN201410271329.1A CN201410271329A CN104004700A CN 104004700 A CN104004700 A CN 104004700A CN 201410271329 A CN201410271329 A CN 201410271329A CN 104004700 A CN104004700 A CN 104004700A
- Authority
- CN
- China
- Prior art keywords
- propionic acid
- dehydrogenase
- engineering bacteria
- propionibacterium jensenii
- overexpression
- 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.)
- Pending
Links
Abstract
The invention discloses a high-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof, and belongs to the field of genetic engineering. A molecular method is adopted. The propionibacterium jensenii engineering bacterium serves as a host, and overexpression is carried out on malate dehydrogenase (mdh) from Klebsiella pneumoniae, or overexpression is simultaneously carried out on glycerol dehydrogenase (gldA) from the Klebsiella pneumoniae. Compared with an original strain, the propionic acid yield of recombination and mixing coexpression glycerol dehydrogenase of overexpression malate dehydrogenase and the propionic acid yield of a recombinant strain of the malate dehydrogenase are 36.09 g/L and 39.43 g/L respectively, and are improved by 33.91 % and 46.3 % respectively than that of the original strain. The method provides new ideas for modifying the propionibacterium jensenii and improving productivity of propionic acid fermentation in the industrial biotechnology.
Description
Technical field
The present invention relates to a kind of high yield propionic acid propionibacterium jensenii engineering bacteria and application thereof, belong to field of genetic engineering.
Background technology
Propionic acid and derivative thereof are very extensive in industrial Application Areas, are mainly used in the aspects such as food antiseptic, feeder reservoir, medicine intermediate are synthetic, agriculture weedkiller is synthetic, organic synthesis intermediate.World's propionic acid overall throughput in 2006,350,000 tons/year of left and right, reaches 500,000 tons/year for 2008.The U.S. is maximum in the world propionic acid Production and consumption state.The actual annual production of China's propionic acid at present only has 200 tons of left and right, can not meet the actual market requirement far away, still needs a large amount of imports to make up domestic vacancy every year.
Propionibacterium jensenii (Propionibacterium jensenii) belongs to propiono-bacterium, is anerobe, is a class G
+, do not produce gemma, do not move, the anerobe of the catalase positive.Bacterium colony presents white, yellow or maroon.Generally, optimal pH is 6.5～7.5, and the temperature of the most suitable growth is 28 ℃～37 ℃.
The production method of propionic acid has chemical synthesis and microbe fermentation method, and chemical synthesis is to take the Chemicals such as oil as raw material, and through heating, pressurizeing, utilizing the propionic acid synthesized method of catalyzer, the method is the main production method of propionic acid in industrial scale.Microbe fermentation method is that bacterium acidi propionici utilizes general nutrition source by self metabolism generation propionic acid under normal temperature, normal pressure.Because microorganism fermentation productions of ethylformic acid can reduce the dependency to Nonrenewable energy resources such as oil, alleviate the pressure that environment is caused, therefore, in the current whole world, face in environmental pollution, the severe situation of energy starved, Progresses of Propionic Acid Production by Microbial Fermentation provides new thinking for propionic acid is synthetic, has obtained more and more investigators' concern simultaneously.But microbial method is produced propionic acid and is still existed cost high, the propionic acid shortcoming such as yield poorly.The main carbon source of propionibacterium is glycerine, yet in its pathways metabolism, propionibacterium jensenii is not high to the utilization ratio of glycerine, thereby causes carbon source to change into the inefficiency of product, finally causes propionic acid to yield poorly.Therefore, need relevant enzyme in the first step in overexpression glycerol metabolism approach be glycerol dehydrogenase gene (gldA) to improve the utilization ratio of propionibacterium to glycerine, finally reach the object of the output that improves propionic acid.
Summary of the invention
Technical problem to be solved by this invention is to provide a kind of high yield propionic acid propionibacterium jensenii engineering bacteria, that to take propionibacterium jensenii (Propionibacterium jensenii) be host, overexpression is from the malate dehydrogenase gene mdh of Klebsiella pneumonia (Klebsiella pneumonia subsp.pneumoniae ATCC 12657), or goes back overexpression from the glycerol dehydrogenase gene gldA of Klebsiella pneumonia simultaneously.
The preferred propionibacterium jensenii JN926 of described propionibacterium jensenii CCTCC No:M 2013071.
The aminoacid sequence of described glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase) is respectively as shown in SEQ ID NO.1, SEQ ID NO.2.
Described glycerol dehydrogenase gene and malate dehydrogenase gene are expressed by expression vector pZGX04.
The construction process of described expression vector pZGX04 is referring to document Zhuge, X., Liu L., Shin, H.D., Chen, R.R., Li, J., Du, G., Chen, J., 2013.Development of a Propionibacterium-Escherichia coli shuttle vector as a useful tool for metabolic engineering of Propionibacterium jensenii, an efficient producer of propionic acid.Appl.Environ.Microbiol.79,4595-4602.
The present invention also provides the method that builds described engineering bacteria, be by the independent overexpression of malate dehydrogenase gene mdh that derives from Klebsiella pneumonia in propionibacterium jensenii, or by the gldA gene co-expressing of mdh and encoding glycerol desaturase in propionibacterium jensenii, to improve the utilization ratio of glycerine, thereby improve glycerol metabolism approach flux, and finally cause the raising of production intensity of propionic acid and output.
The preferred following steps of described method: Klebsiella pneumonia Klebsiella pneumonia subsp.pneumoniae ATCC 12657 whole genome sequences of take are template, pcr amplification obtains or adopts the complete synthesis method of chemistry to obtain mdh object fragment.By flat end, connect and be cloned into plasmid pZGX04 (Zhuge et al., 2013), obtain mdh gene expression plasmid pZGX04-mdh.The recombinant plasmid pZGX04-mdh building proves through DNA sequencing, shows that construction of recombinant plasmid is correct.The method that recombinant plasmid is transformed by electric shock, proceeds to recipient bacterium Propionibacterium jensenii, on SLB substratum, selects immediately, utilizes the method for bacterium colony PCR to screen, and obtains the recombinant bacterium of malate dehydrogenase (malic acid dehydrogenase) overexpression.
Described method is preferred following steps also: Klebsiella pneumonia Klebsiella pneumonia subsp.pneumoniae ATCC 12657 whole genome sequences of take are template, and pcr amplification obtains or adopts the complete synthesis method of chemistry to obtain gldA and mdh object fragment.By flat end, connect and be cloned into plasmid pZGX04 (Zhuge et al., 2013), obtain gldA and mdh gene expression plasmid pZGX04-gldA-mdh.The recombinant plasmid pZGX04-gldA-mdh building proves through DNA sequencing, shows that construction of recombinant plasmid is correct.The method that recombinant plasmid is transformed by electric shock, proceed to recipient bacterium Propionibacterium jensenii, on SLB substratum, select immediately, utilize the method for bacterium colony PCR to screen, obtain the recombinant bacterium of glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase) overexpression.
The present invention also provides a kind of method that propionibacterium jensenii produces propionic acid that improves, adopting above-mentioned propionibacterium jensenii engineering bacteria is starting strain, the seed of standing cultivation 48h in 32 ℃ of anaerobism bottles is proceeded in the fermentor tank that fermention medium is housed with 10% inoculum size, under 32 ℃, 200rpm condition, cultivate, the 3.33mL/h constant speed stream of take between 60-132h adds the glycerine solution that concentration is 300g/L, intermittently logical nitrogen.
Seed culture medium (g/L): Sodium.alpha.-hydroxypropionate 10g, yeast powder 10g, pancreas peptone soybean broth (Trypticase soy broth) 10g, pH7.0.
Fermention medium (g/L): yeast powder 10g, pancreas peptone soybean broth 5g, K
2hPO
42.5g, KH
2pO
41.5g, glycerine 30g, CoCl
28mg, ZnSO
48mg, MgSO
48mg, pH7.0.
Beneficial effect of the present invention: feature of the present invention is to take propionibacterium jensenii as starting strain, utilize Molecular tools to build the recombinant bacterium of a strain overexpression malate dehydrogenase (malic acid dehydrogenase) and the recombinant bacterium of a strain overexpression glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase), by increasing glycerol metabolism flux, cause the raising of propionic acid output.The production intensity of the genetic engineering bacterium of overexpression malate dehydrogenase (malic acid dehydrogenase) and output have improved 33.9% and 33.9% than starting strain respectively, and fed-batch fermentation propionic acid output is brought up to 36.09g/L by original 26.95g/L.The production intensity of the genetic engineering bacterium of overexpression glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase) and output have improved 46.6% and 46.3% than starting strain respectively, and fed-batch fermentation propionic acid output is brought up to 39.43g/L by original 26.95g/L.This transforms propionibacterium jensenii and improve propionic fermentation throughput for industrial biotechnology provides new thinking.
Biomaterial preservation
Propionibacterium jensenii (Propionibacterium jensenii) JN926, is preserved in Chinese Typical Representative culture collection center (CCTCC) on March 6th, 2013, Wuhan, China Wuhan University, and deposit number is CCTCC NO:M 2013071.
Embodiment
The mensuration of dry cell weight (DCW): get a certain amount of bacteria suspension, suitably use UV 7500 type visible spectrophotometers after dilution with dilute hydrochloric acid solution, survey OD value in 600nm place colorimetric, calculate to obtain dry cell weight with dry cell weight typical curve.
The measuring method of propionic acid: high performance liquid chromatography (HPLC)
Instrument: Agilent1200 high performance liquid chromatograph (joining UV-vis detector, differential refraction detector and workstation)
Chromatographic condition: chromatographic column: Agilent ZORBAX SB-Aq post, 5 μ m, 4.6mm * 250mm; Moving phase: 0.138mol/L NaHPO4,1% (v/v) acetonitrile, is adjusted to pH2.0 with phosphoric acid; Flow velocity: 0.8mL/min; Column temperature: 35 ℃; Sample size: 10 μ L; UV-detector wavelength: 210nm.
Sample preparation: 1.5mL fermented liquid is centrifugal 5min under 8000rpm, gets supernatant liquor and moves in 10mL volumetric flask, with 3.68mm/L sulphuric acid soln, is settled to 10mL, and after 0.22 μ m membrane filtration, filtrate is for liquid phase analysis.
The measuring method that glycerol dehydrogenase enzyme is lived:
3mL fermented liquid is centrifugal 5min under 1000rpm, uses 0.5mL K
2hPO
4-KH
2pO
4damping fluid (50mM, pH7.0) is washed once, then uses the damping fluid of same volume resuspended.1000rpm after ultrasonic disruption cell, 4 ℃ of centrifugal 15min, get 0.2mL supernatant, 30mM ammonium sulfate, 0.2M glycerine, 1.2mM NAD is mixed in after 3mL reaction tank, at 340nm, measures the changing value of per minute light absorption value at 30 ℃.Utilization formula below calculates fermentation broth enzyme and lives:
The measuring method that malate dehydrogenase (malic acid dehydrogenase) enzyme is lived:
3mL fermented liquid is centrifugal 5min under 1000rpm, uses 0.5mL K
2hPO
4-KH
2pO
4damping fluid (50mM, pH7.0) is washed once, then uses the damping fluid of same volume resuspended.1000rpm after ultrasonic disruption cell, 4 ℃ of centrifugal 15min, get 0.2mL supernatant, 60mM phosphoric acid salt (pH7.5), 0.2mM NADH, 0.3mM oxaloacetic acid is mixed in after 3mL reaction tank, at 340nm, measures the changing value of per minute light absorption value at 30 ℃.Utilization formula below calculates fermentation broth enzyme and lives:
Embodiment 1 Klebsiella pneumonia gldA gene and the acquisition of mdh gene and the structure of expression plasmid
GldA gene and mdh gene can obtain by chemical total synthesis method, also can obtain by following PCR mode.
(1) take Klebsiella pneumonia Klebsiella pneumonia subsp.pneumoniae ATCC 12657 whole genome sequences is template, and pcr amplification obtains and gldA gene and the mdh gene object fragment of expecting that size conforms to.
(2) during coexpression two gene: it is upper that goal gene is connected to expression vector pZGX04 by flat end, obtains recombinant expression plasmid pZGX04-gldA-mdh.Through sequence verification, prove and connect correctly.
(3) while only crossing expression malate dehydrogenase gene: it is upper that goal gene is connected to expression vector pZGX04 by flat end, obtains recombinant expression plasmid pZGX04-mdh.Through sequence verification, prove and connect correctly.
(4) while only crossing expression glycerol dehydrogenase: it is upper that goal gene is connected to expression vector pZGX04 by flat end, obtains recombinant expression plasmid pZGX04-gldA.Through sequence verification, prove and connect correctly.
PZGX04 is the shuttle vectors between intestinal bacteria-propionibacterium, and in intestinal bacteria, selective marker is penbritin, and in propionibacterium, selective marker is paraxin, and its construction process is shown in document Zhuge et al., 2013.
The Construction and identification of embodiment 2 recombinant bacteriums
Owing to there being restricted modification in propionibacterium jensenii, recombinant plasmid is after e. coli jm109 amplification, first transform intestinal bacteria JM110, the recombinant plasmid electricity extracting from intestinal bacteria JM110 transforms propionibacterium jensenii P.jensenii CCTCC No:M 2013071, is applied in the seed culture medium that contains chlorampenicol resistant.Select immediately transformant and carry out bacterium colony PCR checking, obtain positive transformant, by further separation quality grain, carry out the definite propionibacterium jensenii recombinant bacterium that obtained of sequence verification.
Embodiment 3 recombinant bacteriums with contrast bacterium fermenting experiment
By original bacterium P.jensenii CCTCC NO:M 2013071 and recombinant bacterium P.jensenii CCTCC NO:M2013071-gldA-mdh, P.jensenii CCTCC NO:M 2013071-gldA, P.jensenii CCTCC NO:M2013071-mdh carries out upper tank fed-batch fermentation, fermentation parameter is as shown in table 1, four bacterium are compared: (1) crosses the recombinant bacterium of expressing glycerol dehydrogenase, cross recombinant bacterium and the coexpression glycerol dehydrogenase of expressing malate dehydrogenase (malic acid dehydrogenase), it is 34.62 that the propionic acid output of the recombinant bacterium of malate dehydrogenase (malic acid dehydrogenase) is respectively, 36.09 and 39.43g/L, than starting strain (26.95g/L), improved 28.46, 33.91 and 46.3%, (2) glycerol dehydrogenase of the recombinant bacterium of coexpression glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase) is more alive and malate dehydrogenase (malic acid dehydrogenase) is more obvious than enzyme raising alive than enzyme, the overexpression successful that shows glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase), enzyme is lived and has been improved respectively 6.93 and 2.89 times, (3) cross express glycerol dehydrogenase recombinant bacterium, to cross the production intensity of propionic acid of expressing the recombinant bacterium of malate dehydrogenase (malic acid dehydrogenase) and the recombinant bacterium of coexpression glycerol dehydrogenase and malate dehydrogenase (malic acid dehydrogenase) be 0.152,0.158 and 0.173g/ (Lh).
Table 1 bacterium and the recombinant bacterium fermentation parameter contrast of setting out
Although the present invention with preferred embodiment openly as above; but it is not in order to limit the present invention, any person skilled in the art, without departing from the spirit and scope of the present invention; all can do various changes and modification, so protection scope of the present invention should be with being as the criterion that claims were defined.
Claims (10)
1. a propionibacterium jensenii engineering bacteria for high yield propionic acid, is characterized in that, overexpression is from the malate dehydrogenase gene mdh of Klebsiella pneumonia.
2. engineering bacteria according to claim 1, is characterized in that, has gone back coexpression from the glycerol dehydrogenase gene gldA of Klebsiella pneumonia simultaneously.
3. engineering bacteria according to claim 1 and 2, is characterized in that, the aminoacid sequence of malate dehydrogenase (malic acid dehydrogenase) is as shown in SEQ ID NO.2.
4. engineering bacteria according to claim 2, is characterized in that, the aminoacid sequence of glycerol dehydrogenase is as shown in SEQ ID NO.1.
5. engineering bacteria according to claim 1 and 2, is characterized in that, propionibacterium jensenii (Propionibacterium jensenii) the JN926 CCTCC NO:M 2013071 of take is host.
6. engineering bacteria according to claim 1 and 2, is characterized in that, take plasmid pZGX04 as expression vector.
7. a method that builds engineering bacteria described in claim 1 or 2, it is characterized in that, goal gene fragment is connected and is cloned into plasmid pZGX04 by flat end, and the recombinant plasmid building proceeds to propionibacterium jensenii, obtains the recombinant bacterium of corresponding gene overexpression through screening verification.
8. an application rights requires the method that described in 1 or 2, engineering bacteria is produced propionic acid, it is characterized in that, the seed of standing cultivation 48h in 32 ℃ of anaerobism bottles is proceeded in the fermentor tank that fermention medium is housed, under 32 ℃, 200rpm condition, cultivate, the 3.33mL/h constant speed stream of take between 60-132h adds the glycerine solution that concentration is 300g/L, intermittently logical nitrogen.
9. method according to claim 8, is characterized in that, the cultivation seed culture medium used of described seed consists of: Sodium.alpha.-hydroxypropionate 10g, and yeast powder 10g, pancreas peptone soybean broth 10g, pH7.0, is dissolved in 1L water.
10. method according to claim 8, is characterized in that, described fermention medium consists of: yeast powder 10g, Tryptones is meat soup 5g mostly, K
2hPO
42.5g, KH
2pO
41.5g, glycerine 30g, CoCl
28mg, ZnSO
48mg, MgSO
48mg, pH7.0, is dissolved in 1L water.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410271329.1A CN104004700A (en) | 2014-06-17 | 2014-06-17 | High-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201410271329.1A CN104004700A (en) | 2014-06-17 | 2014-06-17 | High-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN104004700A true CN104004700A (en) | 2014-08-27 |
Family
ID=51365610
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201410271329.1A Pending CN104004700A (en) | 2014-06-17 | 2014-06-17 | High-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN104004700A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104357497A (en) * | 2014-10-22 | 2015-02-18 | 江南大学 | Method for improving yield of propionic acid of acid production propionibacterium |
Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101415830A (en) * | 2006-03-31 | 2009-04-22 | 莱斯大学 | Anaerobic fermentation of glycerol |
| CN102031227A (en) * | 2010-11-25 | 2011-04-27 | 江南大学 | Saccharomyces cerevisiae genetic engineering strain for producing fumaric acid as well as construction method and application thereof |
| CN102286388A (en) * | 2011-06-21 | 2011-12-21 | 江南大学 | Construction method and use of malic acid producing candida glabrata engineering strain |
| CN102286387A (en) * | 2011-06-21 | 2011-12-21 | 江南大学 | Construction method and use of fumaric acid producing candida glabrata engineering strain |
| CN102933715A (en) * | 2009-10-29 | 2013-02-13 | 韩国生命工学研究院 | Novel method for producing 3-hydroxypropionic acid from glycerol |
| CN103361285A (en) * | 2013-05-21 | 2013-10-23 | 江南大学 | Propionibacterium jensenii and application thereof |
| CN103703122A (en) * | 2011-07-29 | 2014-04-02 | 三井化学株式会社 | Microorganism having carbon dioxide fixation pathway introduced thereinto |
-
2014
- 2014-06-17 CN CN201410271329.1A patent/CN104004700A/en active Pending
Patent Citations (7)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101415830A (en) * | 2006-03-31 | 2009-04-22 | 莱斯大学 | Anaerobic fermentation of glycerol |
| CN102933715A (en) * | 2009-10-29 | 2013-02-13 | 韩国生命工学研究院 | Novel method for producing 3-hydroxypropionic acid from glycerol |
| CN102031227A (en) * | 2010-11-25 | 2011-04-27 | 江南大学 | Saccharomyces cerevisiae genetic engineering strain for producing fumaric acid as well as construction method and application thereof |
| CN102286388A (en) * | 2011-06-21 | 2011-12-21 | 江南大学 | Construction method and use of malic acid producing candida glabrata engineering strain |
| CN102286387A (en) * | 2011-06-21 | 2011-12-21 | 江南大学 | Construction method and use of fumaric acid producing candida glabrata engineering strain |
| CN103703122A (en) * | 2011-07-29 | 2014-04-02 | 三井化学株式会社 | Microorganism having carbon dioxide fixation pathway introduced thereinto |
| CN103361285A (en) * | 2013-05-21 | 2013-10-23 | 江南大学 | Propionibacterium jensenii and application thereof |
Non-Patent Citations (2)
| Title |
|---|
| XIN ZHUGE等: ""Development of a Propionibacterium-Escherichia coli shuttle vector as a useful tool for metabolic engineering of Propionibacterium jensenii,an efficient producer of propionic acid"", 《APPL ENVIRON MICBIOL》 * |
| 祝琳琪: ""厌氧微生物高效生产丙酸和丙烯酸的研究"", 《中国博士学位论文全文数据库-基础科学辑》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN104357497A (en) * | 2014-10-22 | 2015-02-18 | 江南大学 | Method for improving yield of propionic acid of acid production propionibacterium |
| CN104357497B (en) * | 2014-10-22 | 2018-04-27 | 江南大学 | A kind of method for putting forward high acid Propionibacterium propionic acid yield |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN108060114B (en) | A kind of Escherichia coli of fermenting and producing l-Alanine and its application | |
| CN105567622B (en) | A kind of application in recombination bacillus coli and synthesis 3- hydracrylic acid | |
| CN103898035B (en) | Produce the recombinant escherichia coli strain of Beta-alanine and construction process thereof and application | |
| CN105400796A (en) | Gene for adjusting and controlling production of long-chain diacid and application of gene | |
| CN102994539A (en) | Method for enhancing expression of corynebacterium crenatum NAD kinase to improve production capacity of strain L-arginine under high-low oxygen supply conditions | |
| CN107129959A (en) | Produce the construction method and its application of (R) 3-hydroxy-2-butanone engineering strain | |
| CN104878024A (en) | Recombinant Escherichia coli construction and (S)-2-hydroxy-3-phenylpropionic acid synthesis method | |
| CN104845893B (en) | One plant of rhodotorula mucilaginosa and its application in the red astaxanthin in fermenting and producing sea | |
| CN104046586B (en) | One strain gene engineering bacterium and the application in producing (2R, 3R)-2,3-butanediol thereof | |
| CN107201374A (en) | The construction method of the butanediol high-yielding engineering bacterial strains of optical voidness meso 2,3 and application | |
| CN104673736A (en) | Method for improving gluconobacter oxydans for producing 2-keto-L-gulconic acid | |
| CN100558884C (en) | A kind of acid-producing Klebsiella bacterium and application thereof | |
| CN104789516B (en) | A kind of genetic engineering bacterium of cytoalgae 6803 for producing trihydroxy propionic acid and construction method and application | |
| CN108641992A (en) | The riboflavin-produced engineering bacteria of high temperature and its construction method and application | |
| CN105483171B (en) | A kind of raising ubiquinone10The production method of industrial output | |
| CN104745545B (en) | A kind of method of efficiently production L dglutamic oxidases | |
| CN104004700A (en) | High-yield propionic acid propionibacterium jensenii engineering bacterium and application thereof | |
| CN104357497B (en) | A kind of method for putting forward high acid Propionibacterium propionic acid yield | |
| CN106868033A (en) | The Corynebacterium glutamicum strain of high yield chiral D () 3-hydroxy-2-butanone and structure and application | |
| CN105368884A (en) | Recombinant bacterium with function of pyruvate production intensity improvement and application of recombinant bacterium with function of pyruvate production intensity improvement | |
| CN105219672A (en) | A kind of extensive method preparing methane-oxidizing bacteria | |
| CN109929854A (en) | One plant is overexpressed the engineering bacteria and its construction method that catabolite controls albumin A encoding gene | |
| CN107201375A (en) | Produce the construction method and its application of (R, R) 2,3 butanediol engineering strain | |
| CN108588108B (en) | Preparation method and application of bacillus for efficiently metabolizing glycerol | |
| CN109563471A (en) | Generate the bacterium bacterial strain of 2,3- butanediol and other metabolins |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20140827 |