CN105950731A - Method for analyzing and researching butanol tolerance key genes of Escherichia coli on basis of gene chips - Google Patents
Method for analyzing and researching butanol tolerance key genes of Escherichia coli on basis of gene chips Download PDFInfo
- Publication number
- CN105950731A CN105950731A CN201610352528.4A CN201610352528A CN105950731A CN 105950731 A CN105950731 A CN 105950731A CN 201610352528 A CN201610352528 A CN 201610352528A CN 105950731 A CN105950731 A CN 105950731A
- Authority
- CN
- China
- Prior art keywords
- gene
- butanol
- coli
- butanol tolerance
- yghw
- 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.)
- Withdrawn
Links
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6876—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes
- C12Q1/6888—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms
- C12Q1/689—Nucleic acid products used in the analysis of nucleic acids, e.g. primers or probes for detection or identification of organisms for bacteria
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12Q—MEASURING OR TESTING PROCESSES INVOLVING ENZYMES, NUCLEIC ACIDS OR MICROORGANISMS; COMPOSITIONS OR TEST PAPERS THEREFOR; PROCESSES OF PREPARING SUCH COMPOSITIONS; CONDITION-RESPONSIVE CONTROL IN MICROBIOLOGICAL OR ENZYMOLOGICAL PROCESSES
- C12Q1/00—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions
- C12Q1/68—Measuring or testing processes involving enzymes, nucleic acids or microorganisms; Compositions therefor; Processes of preparing such compositions involving nucleic acids
- C12Q1/6844—Nucleic acid amplification reactions
- C12Q1/686—Polymerase chain reaction [PCR]
Abstract
The invention discloses a method for analyzing and researching butanol tolerance key genes of Escherichia coli on the basis of gene chips, and belongs to the technical field of bioengineering. The method has the advantages that transcription level difference between genomes of the Escherichia coli E. coli JM109/pHACMB8 and genomes of control strains under butanol stress conditions is particularly analyzed by the aid of gene chip technologies, the two genes yibT and yghW which are closely related to the butanol tolerance of the Escherichia coli and major metabolic pathways related to the butanol tolerance are obtained by means of authentication, and theoretical bases can be provided for further illustrating butanol tolerance mechanisms of microbial strains.
Description
Technical field
The invention belongs to technical field of bioengineering, relate to a kind of based on gene microarray analysis research escherichia coli butanol tolerance key gene and the method for relevant metabolic pathway, be applied to probing into of escherichia coli butanol tolerance mechanism.
Background technology
Most of organic solvents have inhibitory action to growth of microbial cells.They can destroy cell membrane, changes intracellular pH, causes intracellular ion, the loss of metabolite and the false folding of albumen, causes cell damage the most dead, and this has had a strong impact on microbial strains application in industrial circle.Therefore, the Organic Solvent Tolerant mechanism of further investigation microbial cell, for microbial strains organic solvent tolerance, and have great importance in the application of industrial circle.
Gene chip is the High biotechnology developed rapidly in recent years, has the advantages such as efficient, quick, accurate, sensitive, can the function of bar genes thousands of to organism simultaneously study, the complex network relation of special gene's expression and control.2003, Hiroyuki et al. uses biochip technology to find, and the process LAN of marA gene can significantly improve colibacillary organic solvent tolerance, its reason is probably the transcription factor that the MarA albumen of marA coding is acrAB-tolC efflux pump, its process LAN contributes to intracellular organic solvent is excreted (Hayashi S, et al.Journal of Bioscience and Bioengineering.2003,95 (4): 379-383).2007, Hirasawa etc. use this technology for detection two Accharomyces cerevisiae gene expression profile under 5% (v/v) ethanol is coerced, by to Cluster Analysis of Gene Expression Data, find that the expression of tryptophan biosynthesis related gene is favorably improved alcohol resistance (the Hirasawa T of yeast cells, et al.Journal of Biotechnology, 2007,131 (1): 34-44.).But, the butanol tolerance mechanism about microbial strains still needs to be studied further.
Summary of the invention
It is an object of the invention to the key gene relevant based on gene microarray analysis research escherichia coli butanol tolerance and metabolic pathway, by cell membrane fat acid constituents is analyzed, explore the mechanism of action of key gene.The main metabolic pathway relevant to escherichia coli butanol tolerance includes, glyoxalic acid dicarboxylic acids, propanoic acid, acetone acid, oxidative phosphorylation, sugar and amino acid metabolism approach, and abc transport system.Knocking out of gene yibT, yghW is favorably improved colibacillary butanol tolerance, and its tolerance mechanism is relevant with the increase of cell membrane unsaturated fatty acid content.The present invention provides experimental basis for building Organic Solvent Tolerant bacterial strain, and the butanol tolerance mechanism for explaination microorganism further is provided fundamental basis.
Technical scheme:
After the butanolatolerant bacterial strain E.coli JM109/pHACMB8 obtained in early-stage Study and control strain are cultivated 1.5h under 0.8% (v/v) butanol concentration, extracting experimental strain and the RNA of control strain respectively, each sample comprises three repetitions biologys respectively.Use gene chip (Agilent custom chip design ID:020097) technology respectively the gene transcription level of mutant and control strain to be detected, process through data normalization and finally draw butanolatolerant mutant differential expression situation of gene compared with control strain.According to the screening parameter of gene chip differential gene, screen fold differences>2, the difference expression gene of p-value<0.05.Result obtains 329 difference expression genes (wherein 197 gene transcription levels raise, and 132 gene transcription levels are lowered) altogether.Gene chip related data has been submitted in GEO public database, numbered GSE79305.
Utilize KEGG data base that the characteristic distributions of differential gene and the function of gene are analyzed.
Use engineered method (gene overexpression and gene knockout) that the function of 15 up-regulated genes and 14 down-regulated genes is studied.
By measuring cell membrane fat acid content, the tolerance mechanism of butanol tolerance key gene yibT, yghW is studied.
Beneficial effects of the present invention:
(1) present invention uses biochip technology to be analyzed the transcript profile of butanolatolerant bacterial strain E.coli JM109/pHACMB8 and control strain, (fold differences is all more than 2 times to obtain 329 difference expression genes, p-value < 0.05), wherein 197 gene transcription levels raise, and 132 gene transcription levels are lowered.
(2) use KEGG data base that difference expression gene has carried out function and metabolic pathway enrichment is analyzed.Find that the metabolic pathway relevant to butanol tolerance has glyoxalic acid dicarboxylic acids metabolism, propanoic acid metabolism, metabolism of pyruvate, oxidative phosphorylation, carbohydrate metabolism, amino acid metabolism and abc transport system.
(3) use engineered method that the function of 15 up-regulated genes and 14 down-regulated genes is verified.Result shows: the process LAN of up-regulated gene glcF, gcl, yhaR, ybbQ and tdcE is favorably improved colibacillary butanol tolerance；And after being knocked out by down-regulated gene yibT and yghW, the butanol tolerance of bacterial strain also increases.The tolerance mechanism of yibT, yghW is carried out research and finds that the cell membrane unsaturated fatty acid content of knock-out bacterial strain Δ yibT and Δ yghW dramatically increases.
The present invention provides experimental basis for building Organic Solvent Tolerant bacterial strain, and the butanol tolerance mechanism for explaination microorganism further is provided fundamental basis.
Accompanying drawing explanation
The SDS-PAGE collection of illustrative plates of Fig. 1 up-regulated gene process LAN bacterial strain.
Fig. 2 down-regulated gene knock-out bacterial strain and control strain bacterium colony formation efficiency comparison under organic solvent pressure.
Detailed description of the invention
Embodiment 1 butanolatolerant bacterial strain and the extraction of control strain total serum IgE and microarray data process
After butanolatolerant strain Escherichia coli E.coli JM109/pHACMB8 and control strain are cultivated 1.5h under 0.8% (v/v) butanol concentration, 4 DEG C, 8000r min-1 collects thalline, quickly grinds thalline with liquid nitrogen, with Trizol reagent eluting and collect intracellular organic matter.By using QIAGEN RNeasy mini kit and RNase-Free DNase Set test kit that total serum IgE is purified after electrophoresis quality inspection.Each sample comprises three repetitions biologys respectively.Sample storage is delivered in dry ice Shanghai Biotechnology Corporation and carries out gene chip hybridization experiment.
The chip completing hybridization uses Agilent Microarray Scanner to be scanned, and reads data with Feature Extraction software 10.7, finally uses Gene Spring Software 11.0 to be normalized data.Use molecular network annotation system (Bioknow-Molnet) by with KEGG PATHWAY data base (http://www.genome.ad.jp/kegg/pathway.html) phase coupling, the differential gene in gene chip can be carried out functional annotation and metabolic pathway enrichment analyze.Gene sets relevant or close for one group of function is sorted out, it is thus achieved that metabolic pathway closely-related with butanolatolerant.KEGG data base the results are shown in Table 1 to the enrichment of differential gene.
The enrichment of table 1 gene chip differential gene KEGG metabolic pathway is analyzed
The structure of the process LAN bacterial strain of embodiment 2 up-regulated gene
E.coli JM109/pQE80L is inoculated in LB/Kan fluid medium, after 37 DEG C are cultivated 10h, extracts pQE80L plasmid, and with restricted enzyme Bam HI and Hind III after 37 DEG C of double digestion 3h, post reclaims to obtain the linearization plasmid of purification.With E.coli JM109 genome as template, carrying out PCR with the up-regulated gene primer with same restriction enzyme site, PCR system is as follows:
PCR program: 95 DEG C of 5min, 98 DEG C of 10s, 55 DEG C of 15s, 72 DEG C of 2min 30s react 30 circulations, 72 DEG C of 10min, 16 DEG C of 30s.
The PCR primer fragment PCR primer obtaining associated upregulated gene is reclaimed test kit be purified, by Bam HI and the recovery fragment of Hind III double digestion up-regulated gene, according to linked system description, in system, add carrier and the fragment of debita spissitudo.16 DEG C overnight connect, and linked system is as follows:
The reactant liquor overnight connected is gone to E.coli JM109 competent cell by chemical heat robin, cultivates 1.5h afterwards, coat LB/Kan flat board, cultivate 12h, the single bacterium colony on picking flat board, carry out bacterium colony PCR checking with pQE universal primer for 37 DEG C.Being inoculated in by positive clone strain in 30mL LB/Kan fluid medium, 37 DEG C, 120rpm cultivates the IPTG adding final concentration of 0.2mmol L-1 when being about 0.6 to OD660, and 30 DEG C, 120rpm induces 4h.8800 × g is centrifuged 10min, abandons supernatant and collects wet thallus, with 10mL PBS suspension cell (pH 7.0,20mmol L-1), and carries out ultrasonication 15min (ultrasonic 1s, intermittently 3s) in Ultrasonic Cell Disruptor.Cell suspending liquid after broken is carried out SDS-PAGE analysis, and result is as shown in Figure 1.Wherein, the albumen of these 11 gene expressions of glcF, gcl, glcD, glcB, glcG, yeiN, yhaR, ybbQ, tdcE, tdcD and yeiC has obvious band in relevant position.But, the albumen that glcA, ECs0572, hyi and adiY express does not has obvious process LAN band.Gene glcA and ECs0572 expresses glycolic transport protein and allantoin permease respectively, belongs to inner membrane protein.After the two protein overexpression, cellular metabolism is unbalance, and strain growth is all suppressed, and the process LAN situation of albumen therefore cannot be detected with albumin glue.Additionally, hyi and adiY is separately encoded the activating transcription factor of hydroxypyruvate isomerase and arginine decarboxylase.Owing to the two enzyme has substrate dependency, the most just can be expressed.
The structure of the knock-out bacterial strain of embodiment 3 down-regulated gene
Extract the plasmid of E.coli BW25113/pKD46 bacterial strain, turn in E.coli JM109 according to common thermal transitions method, owing to pKD46 belongs to Thermo-sensitive plasmid, therefore single bacterium colony is had to occur 30 DEG C of cultivations to Amp flat board, single bacterium colony on picking flat board, in LB/Amp fluid medium, adds the arabinose that 300 μ L concentration are 3mol L-1 in 30mL culture medium simultaneously.When OD660 is about 0.8～1.0, preparation E.coli JM109/pKD46 electricity turns competence.Utilize gene knockout primer (deriving from upstream region of gene-50 to be knocked out to downstream+50 district), with pKD13 plasmid as template, carry out PCR with PrimerSTAR high-fidelity enzyme, knock out genetic fragment to obtain with homology arm.PCR primer is separated by electrophoresis after purification through nucleic acid gel, according to conventional electricity carryover sequence, is turned by the genetic fragment electricity with homology arm and turns in competent cell in above-mentioned E.coli JM109/pKD46 electricity.Coating LB/Kan flat board, cultivates to single bacterium colony generation for 37 DEG C.Single bacterium colony that the above-mentioned electricity of picking grows after turning, carries out bacterium colony PCR checking with checking primer (deriving from upstream region of gene-75 to be knocked out to-50th district and downstream+50 to+75th district) and Kan universal primer.To verify that the successful positive strain of homologous recombination is chosen in LB/Kan fluid medium, 37 DEG C of incubated overnight, transfer afterwards in fresh LB/Kan culture medium, when OD660 is about 0.8～1.0, the electricity preparing corresponding homologous recombination bacterial strain turns competence.The pCP20 plasmid electricity of extraction is turned in corresponding competent cell, is coated with LB/Cm flat board.30 DEG C of overnight incubation, for the elimination of Kan resistance fragments.Single bacterium colony on rear picking flat board, in LB fluid medium, cultivates 12h for 42 DEG C.Picking one ring culture fluid carries out plate streaking, continues 42 DEG C of cultivations, for the elimination of pCP20 plasmid.Choosing 9 single bacterium colonies on flat board, dibbling is in LB, LB/Kan, LB/Amp solid plate respectively.If bacterium colony only grows on LB flat board, and the longest on LB/Kan, LB/Amp flat board, show thirst quenching success.Simultaneously with checking primer further to only on LB flat board growth single bacterium colony carry out bacterium colony PCR checking, it is thus achieved that PCR stripe size at about 250bp, show thirst quenching success.
The butanol tolerance checking of embodiment 4 bacterial strain
The butanol tolerance checking of up-regulated gene process LAN bacterial strain: the process LAN bacterial strain successfully constructed and control strain E.coli JM109/pQE-80L are connected to incubated overnight in LB/Kan fluid medium, (30 DEG C, 120rpm) it are forwarded in the fresh LB/Kan culture medium containing final concentration of 0.2mmol L-1IPTG by the switching amount of 1% (v/v).Add 0.8% when OD660 about 0.8 butanol of (v/v), measures the concentration (OD660) of each bacterial strain after cultivating 8h.Result is as shown in table 2.Wherein, under 0.8% (v/v) butanol pressure, the 6 strain process LAN strain growth situations carrying recombiant plasmid pQE80L-glcF, pQE80L-gcl, pQE80L-yhaR, pQE80L-glcB, pQE80L-ybbQ and pQE80L-tdcE are all better than control strain.Cell concentration improves 46.63% than control strain respectively, and 52.90%, 39.56%, 18.69%, 13.52%, 46.36%.
The butanol tolerance checking of table 2 up-regulated gene process LAN bacterial strain
The butanol tolerance checking of down-regulated gene knock-out bacterial strain: the bacterial strain of successful knockout is connected to incubated overnight in LB fluid medium, and next day is forwarded in fresh LB by the switching amount of 1% (v/v).Add 0.8% when OD660 about 0.8 butanol of (v/v), cultivates 1.5h afterwards, takes after appropriate culture fluid dilutes 50 times, then is diluted with 10 times of gradients, draws the bacterium solution dibbling of the 10 each extension rates of μ L respectively in the cup of Oxford.37 DEG C of incubated overnight, the growing state of bacterium colony is as shown in Figure 2.Finding out from the result of knock-out bacterial strain, after down-regulated gene yibT and yghW gene knockout, the butanol tolerance of bacterial strain increases.After knocking out glgS and crcA, the toleration of bacterial strain presents decline phenomenon.Bacterial strain toleration is had little to no effect by knocking out of other genes.
The mensuration of embodiment 5 cell membrane fat acid composition
Learn from else's experience the knock-out bacterial strain Δ yibT of E.coli JM109 and gene yibT and yghW of activated overnight, Δ yghW cell culture fluid, transfer by the switching amount of 1% (v/v) and cultivate to stable phase in 30mL LB culture medium, centrifugal (8800 × g, 4 DEG C, 10min), abandon supernatant and obtain wet thallus.With 10mmol L-1 sodium phosphate buffer (pH 7.4) suspension cell and by above-mentioned pelleted by centrifugation, this step is repeated twice.Taking 300mg wet cell, suspend with 1.6mL sodium phosphate buffer, and add 4mL methanol, 2mL chloroform, three's volume ratio is 0.8:2:1, and room temperature stands 2h, mixes once every 0.5h.In mixed liquor, add 2mL methanol, 2mL chloroform, dilute extracting solution, be centrifuged (8800 × g, 4 DEG C, 10min), take off a layer solution (containing lipid), add the methanol solution containing 5% (v/v) H2SO4,70 DEG C of esterification 2h.Taking 1mL fatty acid methyl ester, add isopyknic pentane and extract three times, add anhydrous sodium sulfate, 8800 × g is centrifuged 2min, is transferred to by supernatant in new centrifuge tube, and pentane liquid nitrogen instrument dries up evaporation, is redissolved by the fatty acid normal hexane of extraction.Use Trace1310GC coupling TSQ8000MSD (Thermo Fisher Scientific, Massachusetts, USA) system, and the sample of derivatization is analyzed by the nonpolar capillary column of HP-5MS (30m × 0.25mm × 0.25 μm).Analysis condition is: 60 DEG C of 2min, after be warming up to 150 DEG C (8 DEG C of min-1), after be warming up to 250 DEG C (3 DEG C of min-1), be then warming up to 280 DEG C (10 DEG C of min-1) keep 5min.Split ratio 5:1.Result is as shown in table 3.
The cell membrane fat acid composition analysis of table 3 gene yibT and yghW knock-out bacterial strain and control strain
As shown in Table 3, the composition that mainly comprises of cell membrane fat acid is C16:1, C16:0, C18:0 and C18:1, and their content accounts for more than the 70% of cell membrane total fatty acid content.Document report C16 and C18 is mainly responsible for mobility and the integrity of cell membrane.We have found that compared with control strain, the content of unsaturated fatty acid in knock-out bacterial strain Δ yibT and Δ yghW, the content of particularly C16:1, C18:1 increased.Wherein C18:1 is as most important unsaturated fatty acid, mainly resists the extraneous noxious substance infringement to cell by the mobility of regulation cell membrane.The change of C16:1 content can affect rigidity and the integrity of cell membrane lipid layer.This result shows, the butanolatolerant mechanism of yibT with yghW is relevant with the constituent change of cell membrane fat acid.
Claims (1)
1. use gene microarray analysis E. coli JM109/pHACMB8 and base of control strain under butanol stress conditions
Because of group transcriptional level difference, identify gene yibT and yghW relevant to butanol tolerance, and relevant metabolic pathway, its
It is characterised by:
(1) gene yibT and yghW encodes putative protein, participates in the regulation and control of cell membrane unsaturated fatty acid content, particularly
C16:1 and C18:1.
(2) comparing with control strain, the butanol tolerance of the knock-out bacterial strain of gene yibT and yghW significantly improves.
(3) butanol tolerance of E. coli JM109/pHACMB8 relates generally to glyoxalic acid dicarboxylic acids, propanoic acid, and third
Keto acid, oxidative phosphorylation, sugar and amino acid metabolism approach, and abc transport system.
(4) E. coli JM109/pHACMB8 carries σ70Mutant gene rpoDM2, the aminoacid sequence of its coding
For SEQ ID NO:1.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610352528.4A CN105950731A (en) | 2016-05-25 | 2016-05-25 | Method for analyzing and researching butanol tolerance key genes of Escherichia coli on basis of gene chips |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201610352528.4A CN105950731A (en) | 2016-05-25 | 2016-05-25 | Method for analyzing and researching butanol tolerance key genes of Escherichia coli on basis of gene chips |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN105950731A true CN105950731A (en) | 2016-09-21 |
Family
ID=56910811
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201610352528.4A Withdrawn CN105950731A (en) | 2016-05-25 | 2016-05-25 | Method for analyzing and researching butanol tolerance key genes of Escherichia coli on basis of gene chips |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN105950731A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107577919A (en) * | 2017-08-21 | 2018-01-12 | 上海派森诺生物科技股份有限公司 | A kind of grand genomic data analysis method based on high throughput sequencing technologies |
| CN113621628A (en) * | 2021-09-27 | 2021-11-09 | 广西科学院 | Application of oxyR gene fragment in host bacterium to resistance to alcohol stress and acid stress |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1580281A (en) * | 2003-08-06 | 2005-02-16 | 宋克 | Gene chip enzyme-method microsphere amplifying label |
| CN104328167A (en) * | 2014-09-17 | 2015-02-04 | 宁夏大学 | Gene chip capable of parallel detection of ten main pathogenic bacteria of cow mastitis and detection method thereof |
-
2016
- 2016-05-25 CN CN201610352528.4A patent/CN105950731A/en not_active Withdrawn
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN1580281A (en) * | 2003-08-06 | 2005-02-16 | 宋克 | Gene chip enzyme-method microsphere amplifying label |
| CN104328167A (en) * | 2014-09-17 | 2015-02-04 | 宁夏大学 | Gene chip capable of parallel detection of ten main pathogenic bacteria of cow mastitis and detection method thereof |
Non-Patent Citations (1)
| Title |
|---|
| HAYASHI S.ET AL: "analysis of organic solvent tolerance in Escherichia coli using gene expression profiles from DNA microarrays", 《J BIOSCI BIOENG》 * |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN107577919A (en) * | 2017-08-21 | 2018-01-12 | 上海派森诺生物科技股份有限公司 | A kind of grand genomic data analysis method based on high throughput sequencing technologies |
| CN113621628A (en) * | 2021-09-27 | 2021-11-09 | 广西科学院 | Application of oxyR gene fragment in host bacterium to resistance to alcohol stress and acid stress |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Shui et al. | Adaptive laboratory evolution of ethanologenic Zymomonas mobilis strain tolerant to furfural and acetic acid inhibitors | |
| Li et al. | Engineering Bacillus subtilis for isobutanol production by heterologous Ehrlich pathway construction and the biosynthetic 2-ketoisovalerate precursor pathway overexpression | |
| CN103667371B (en) | A kind of biological production of Salvianic acidA | |
| US11634720B2 (en) | Yeast producing tyrosol or hydroxytyrosol, and construction methods thereof | |
| He et al. | Transcriptome profiling of Zymomonas mobilis under ethanol stress | |
| Wang et al. | Global metabolomic and network analysis of Escherichia coli responses to exogenous biofuels | |
| Mohagheghi et al. | Improving xylose utilization by recombinant Zymomonas mobilis strain 8b through adaptation using 2-deoxyglucose | |
| Wang et al. | Improvement of stress tolerance and riboflavin production of Bacillus subtilis by introduction of heat shock proteins from thermophilic bacillus strains | |
| Yang et al. | Development and characterization of acidic-pH-tolerant mutants of Zymomonas mobilis through adaptation and next-generation sequencing-based genome resequencing and RNA-Seq | |
| CN103571762B (en) | A kind of Mortierella alpina recombinant gene expression system and construction process thereof and application | |
| WO2020199571A1 (en) | Recombinant yeast, construction method, and application thereof in preparing tyrosol and derivatives | |
| Taweecheep et al. | In vitro thermal and ethanol adaptations to improve vinegar fermentation at high temperature of Komagataeibacter oboediens MSKU 3 | |
| CN104946575A (en) | Escherichia coli expression strain for high production of tyrosol and/or salidroside and icarisid D2 and application of escherichia coli expression strain | |
| Ting et al. | Screened butanol-tolerant Enterococcus faecium capable of butanol production | |
| CN105950731A (en) | Method for analyzing and researching butanol tolerance key genes of Escherichia coli on basis of gene chips | |
| Huang et al. | Furfural-tolerant Zymomonas mobilis derived from error-prone PCR-based whole genome shuffling and their tolerant mechanism | |
| Lou et al. | Development and characterization of efficient xylose utilization strains of Zymomonas mobilis | |
| Wang et al. | An ethanol-tolerant recombinant Escherichia coli expressing Zymomonas mobilis pdc and adhB genes for enhanced ethanol production from xylose | |
| Kalnenieks et al. | Zymomonas mobilis metabolism: Novel tools and targets for its rational engineering | |
| Park et al. | Revealing oxidative pentose metabolism in new Pseudomonas putida isolates | |
| WO2024017318A1 (en) | Ralstonia eutropha having low endotoxin content, and use thereof | |
| CN113493758B (en) | Tyrosol-producing recombinant escherichia coli capable of shortening fermentation period and application thereof | |
| WO2015154209A1 (en) | Bacterial strain capable of producing a large amount of l-alanine and being tolerant to tap water and construction method thereof | |
| US20180057807A1 (en) | Gene modification in clostridium for increased alcohol production | |
| Sun et al. | Comparative genomics and transcriptome analysis of Lactobacillus rhamnosus ATCC 11443 and the mutant strain SCT-10-10-60 with enhanced L-lactic acid production capacity |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| WW01 | Invention patent application withdrawn after publication |
Application publication date: 20160921 |
|
| WW01 | Invention patent application withdrawn after publication |