CN103484388A - Industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways - Google Patents
Industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways Download PDFInfo
- Publication number
- CN103484388A CN103484388A CN201310164960.7A CN201310164960A CN103484388A CN 103484388 A CN103484388 A CN 103484388A CN 201310164960 A CN201310164960 A CN 201310164960A CN 103484388 A CN103484388 A CN 103484388A
- Authority
- CN
- China
- Prior art keywords
- bacterial strain
- xylose
- strain
- saccharomyces cerevisiae
- gene
- 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
Images
Abstract
An industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways is disclosed, and is industrial saccharomyces cerevisiae bacterial strain ZQ5 without any screening marker gene, and belongs to the biological technical field of microbes. The bacterial strain has a classification name of saccharomyces cerevisiae and a registration serial number of CGMCC 7463, and is preserved in China General Microbiological Culture Collection Center, and the preservation unit address is No. 3, No. 1 yard, Beichen West Road, Chaoyang District, Beijing City, and the preservation date is April 11, 2013. The invention also discloses a genetic engineering construction method for the recombination bacterial strain ZQ5, and the method comprises: acquiring of three exogenous xylose initial-utilization genes, construction of a chromosome integrated carrier, and xylose growth experiment of the bacterial strain. Due to the facts that the recombination bacterial strain ZQ5 is capable of utilizing xylose as a carbon source for growth, and the self of the bacterial strain ZQ5 does not carry any antibiotic screening marker genes, the recombination bacterial strain ZQ5 is applicable to industrial production.
Description
Technical field
The invention belongs to field of microbial biotechnology, particularly a strain has the xylose utilization ability, and does not contain the industrial saccharomyces cerevisiae ZQ5 of any selection markers gene.
Background technology
Along with economic fast development, China's energy shortage problem is constantly outstanding.S-generation bio-ethanol, with cellulosic raw material, comprises agricultural crop straw, the fermentative production of ethanol such as forestry waste.With first-generation bio-ethanol difference, be, cellulosic ethanol has been avoided striving grain with the people, with the drawback of grain expropriation of land.China has a large amount of idle cellulosic raw materials, if can fully effectively utilize this natural resource, turns waste into wealth, both can protection of the environment, keep agriculture Sustainable development, and also can reduce dependence and the consumption of China for the Imported oil energy simultaneously.
Lignocellulose starting material basic structure contains three parts: Mierocrystalline cellulose, hemicellulose and xylogen, be interwoven by complicated polysaccharide structures.The cellulosic hydrolysate composition is glucose, and hemicellulose hydrolysate mainly contains wood sugar.Yeast saccharomyces cerevisiae, as the first-selected starting strain of industrial alcohol fermentation, has stronger glucose fermentation ability, but but can not utilize wood sugar growth and fermentation.Therefore, need to comprise bacterium from natural xylose utilization bacterium, the genes involved of clone's xylose metabolism approach in candiyeast and filamentous fungus, the recycling genetic engineering means proceeds to yeast saccharomyces cerevisiae by gene, constructs and can utilize the wood sugar recombinant bacterium of wood sugar as carbon source.Metabolic engineering method commonly used is to express Xylose reductase and the xylose dehydrogenase gene of pichia spp in yeast saccharomyces cerevisiae, is reduced to Xylitol under the recombination microzyme obtained can rely on wood sugar Xylose reductase effect at NADPH, then passes through NAD
+transfer xylulose under the xylitol dehydrogenase oxidation relied on, through the xylulokinase phosphorylation, form X 5P, enter the yeast saccharomyces cerevisiae metabolism.
Lot of domestic and international investigator is absorbed in the research that the wood sugar recombinant bacterium builds in decades, but the enzyme that adopts multi-copy vector to improve the xylose metabolism approach is lived and is expressed more, this class carrier is mostly the free plasmid carrier, need antibiotic-screening, less stable, and easily lose copy, can not be in industrial stable application.Also have and adopt single copy integrative vector foreign gene-carrying fragment, but also carry the antibiotic-screening marker gene simultaneously, or utilize the auxotroph carrier, as tryptophane, leucine, the defective typies such as Histidine are as selection markers, but this class selection markers carrier can't be applicable to large-scale industrial production application, and the saccharomycetic growth of auxotroph is affected.The pAUR135 integrating expression vector that the present invention adopts, not only avoided utilizing microbiotic to select, but genetic stability can also automatically be deleted the resistance fragment after completing integration, only retains the external source fragment on genome, is more suitable for for large-scale commercial production.
Summary of the invention
The object of the invention is to build a strain and can utilize the restructuring an industrial strain of S.cerevisiae strain of wood sugar as carbon source, thereby more can utilize wood sugar in the cellulosic material hydrolyzed solution as carbon source.
The present invention relates to a strain chromosomal integration and express an industrial strain of S.cerevisiae strain of xylose metabolism approach, and an industrial strain of S.cerevisiae strain ZQ5 that does not contain any selection markers gene, this strain classification called after Saccharomyces cerevisiae(yeast saccharomyces cerevisiae), registering on the books of bacterial strain is numbered CGMCC No.7463, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, the depositary institution address is No. 3, Yard 1, BeiChen xi Road, Chaoyang District, Beijing City, and preservation date is on April 11st, 2013.Described bacterial strain contains the xylose metabolism approach, utilizes the wood sugar growth.Described bacterial strain contains three xylose utilization gene Xylose reductase gene PsXR, wood sugar desaturase PsXDH, xylulokinase gene ScXK, the first two gene is from pichia stipitis Pichia stipitis JCM10742, and last gene is from laboratory pattern Saccharomyces Cerevisiae in S 288c.The GenBank accession number of gene order is respectively X59465, AF127801.1, NM_001181323.PGK1 promoter sequence GenBank accession number is FJ415226.1, and CYC1 terminator sequence GenBank accession number is EF210198.1.Adopt the 16 karyomit(e) YPRCdelta15 of yeast saccharomyces cerevisiae, LTR (Long Tandem Repeat) site is integrated, and selects its upstream 700bp and downstream 650bp sequence to carry out Homologous integration as homology arm.
The present invention adopts precious biological (Dalian) pAUR135 of company carrier (article No. D3604) to carry out the integrative gene expression of xylose metabolism gene, the DNA sequence dna that pAUR135DNA can utilize carrier itself comprises that by the carrier part sequence in the yeast saccharomyces cerevisiae transformant microbiotic selective marker AUR1-C gene (gives yeast aureobasidin A ureobasidin A, the AbA resistance) remove, thereby realize the recycle of antibiotic marker.On this carrier with GIN11M86 gene overexpression under Gal10 promotor and semi-lactosi effect, can use the host lethal, therefore can be used for the screening that carrier is removed the type recombinant chou, obtain clone's (only containing goal gene, fully without any other fragments of external source) of AbA responsive type.
Useful achievement of the present invention is: the yeast saccharomyces cerevisiae wood sugar recombinant bacterium ZQ5 in the present invention can utilize wood sugar for carbon source for growth.
The accompanying drawing explanation
Each fragment PCR amplification of Fig. 1 pZQ5.
Fig. 2 pZQ5 vector construction of connecting.
Fig. 3 positive transformant PCR identifies.
Fig. 4 wood sugar recombinant bacterium ZQ5 is at the 50g/l wood sugar, fermentation results in 100g/l glucose.
Embodiment
Embodiment 1: contain structure and the conversion of the industrial saccharomyces cerevisiae of the initial pathways metabolism gene of wood sugar
Three wood sugars that the present invention relates to initially utilize gene order to come from the NCBI public database, and wherein the accession number of PsXR gene (EC:1.1.1.2) is X59465; The accession number of PsXDH gene (EC1.1.1.9) is AF127801.1; The accession number of ScXK gene (EC2.7.1.17) is NM_001181323.The promotor of PsXR and PsXDH gene is all used the PGK1 promotor, ScXK is used the ADH1 promotor, three genes are all as terminator with the CYC1 terminator, and three gene series connection are connected in chromosomal integration vector pAUR-PGK1p-PsXR-CYC1t-PGK1p-PsXDH-CYC1t-ADH1p-ScXK-CYC1t, the upstream and downstream of this box gene adds homology arm sequence 20up and 20down simultaneously, connect together with different restriction endonuclease sites respectively between each fragment, jointly insert yeast the 16 karyomit(e) YPRCdelta15 site.The microbiotic selected marker of this carrier is penbritin (intestinal bacteria) and golden load element (yeast saccharomyces cerevisiae), after the linearized vector electricity is proceeded to Saccharomyces Cerevisiae in S .cerevisiae4126, gold load element selection markers gene can be automatically deleted, therefore exist without any antibiotic-screening marker gene on the karyomit(e) of the ZQ5 recombinated.
1.1 genes of brewing yeast group DNA extraction
(1) yeast liquid of incubated overnight is centrifugal, 12000rpm, 2min, remove supernatant;
(2) add 480 μ L TE solution (pH8.0) to precipitation, 20 μ L lysozyme(2mg/mL), after mixing, concussion puts into 37 ℃ of shaking tables 1.5 hours;
(3) add appropriate RNase A, again put into 37 ℃ of shaking tables 0.5 hour;
(4) take out from shaking table, add 50 μ L20%SDS solution, 5 μ L Proteinase Ks (PK concentration is 20 μ g/mL) mix concussion, more than placing 1h in 55 ℃ of water-baths;
(5) centrifugal pipe is covered to the liquid collecting that adheres to the pipe end; Add 500 μ L phenol: chloroform: second amylalcohol (25:24:1), after concussion mixes, centrifugal 10 minutes of 12000rpm, get supernatant liquor, turns new pipe;
(6) add the equal-volume Virahol, put into-20 ℃ of refrigerators more than 1 hour, precipitation DNA;
(7) 12000rpm is centrifugal 10 minutes, removes supernatant liquor, adds 1mL70% ethanol, washes precipitation 1-2 times, and centrifugal 8 minutes of 12000rpm, abandon supernatant;
Add TE(pH8.0 after (8) 37 ℃ of dryings) 50 μ L dissolving DNAs ,-20 ℃ save backup.
1.2PCR amplification target gene band
Using Pichia stipitis JCM10742DNA as template amplification gene XYL1 and XYL2, using S.cerevisiae S288c DNA as template amplification gene XYKS, using S.cerevisiae6525DNA as the upstream and downstream homology arm sequence of template amplification gene, 20up, 20down.Primer sequence is as follows:
Table 1.2-1 gene amplification primer information
PCR reaction system following (50 μ L):
The PCR response procedures arranges:
After PCR finishes, whether product detects the segment size with agarose gel electrophoresis and meets.
1.3 target segment purifying
Phusion high-fidelity enzymatic amplification segment is flat end, need to add that polyA could be connected with the T carrier, thus add again the polyA tail after the segment purifying, in order to avoid the polyA tail added is cut by the high-fidelity enzyme.Purification reaction carries out with Omega test kit and TIANGEN test kit.
1.4 the target segment adds A tail (20 μ L system)
Be placed in 72 ℃ of water-baths and take out after reaction 20-30min, be connected with carrier after the test kit purifying.
1.5 be connected (10 μ L) with carrier
Reaction times 2-8 hour in connecting instrument, temperature of reaction is 22 ℃.
1.6 connect product transforms in bacillus coli DH 5 alpha
1.6.1 competent escherichia coli cell preparation
(1) inoculate bacillus coli DH 5 alpha in 10mL LB liquid nutrient medium, 37 ℃, the 220rpm overnight incubation;
(2) in the bacterium liquid of the ratio of 1:100 switching incubated overnight to the fresh LB liquid nutrient medium of 50mL, 37 ℃, 220rpm cultivates 3-4h, approximates 0.6 to OD600;
(3) bacterium liquid is proceeded in the 50mL of precooling on ice centrifuge tube, place 30min on ice, 4 ℃, the centrifugal 5min of 4000rpm;
(4) abandon supernatant, with the 0.1mol/L CaCl2 solution 15mL suspension cell of precooling, place 30min on ice, 4 ℃, the centrifugal 5min of 4000rpm;
(5) repeat above-mentioned steps;
(6) with the 0.1mol/L CaCl2 solution 2mL suspension cell of precooling, add 30% glycerine of 2mL precooling, mix gently, be distributed into the aliquot of 200 μ L ,-76 ℃ are frozen.
1.6.2 connect the conversion of product
(1) get 200 μ L competent cells from-76 ℃ of refrigerators, melt on ice;
(2) will connect product and all add light finger bomb tube bottom in competent cell, place 30min on ice;
(3) cell is placed in to 42 ℃ of water-bath thermal shocks and is placed in rapidly afterwards cooled on ice 1.5min in 90 seconds, and then thermal shock 90 seconds, cooled on ice 10min;
(4) Xiang Guanzhong adds 500 μ L LB liquid nutrient mediums (not containing microbiotic), mixes rear 37 ℃ of shaking culture 45 minutes to 1 hour;
(5) the centrifugal 5min of 4000rpm, remove supernatant, surplus solution is coated containing corresponding antibiotic screening dull and stereotyped upper (penbritin final concentration 100mg/mL), face up and place half an hour, be inverted culture dish after bacterium liquid dries fully, cultivate 16-18 hour for 37 ℃;
(6) choosing bacterium colony is identified.
1.6.3 transformant plasmid extraction (solution preparation method used derives from Takara(Dalian) website)
(1) picking transforms being cloned in the fresh LB substratum that is added with corresponding microbiotic (penbritin final concentration 100mg/mL) on flat board, 37 ℃, 220rpm overnight incubation;
(2) get 3ml bacterium liquid in Eppendof pipe l.5mL, the centrifugal l min of 12000rpm, abandon supernatant, the collecting precipitation thalline;
(3) precipitation is resuspended in the solution S olution I of 100 μ L precoolings on ice to the vortex concussion;
(4) add 200 μ l Solution II, put upside down gently centrifuge tube 5 times, solution is mixed;
(5) add the solution S olution III of 150 μ L precoolings on ice, turn upside down for several times, centrifuge tube is placed on ice, place 5min;
(6) 4 ℃, the centrifugal 10min of 12000rpm, stay supernatant;
(7) add equal-volume (approximately 500 μ l) phenol: chloroform: primary isoamyl alcohol (25:24:1), fully concussion, the centrifugal 10min of 12000rpm, transfer to supernatant in new pipe;
(8) add the Virahol of equal-volume precooling, turn upside down and fully mix, in-20 ℃ of placement 30min, the centrifugal 10min of 12000rpm, abandon supernatant;
(9) by 1mL70% washing with alcohol precipitation, the centrifugal 2min of 12000rpm, remove supernatant liquor, can repeat this washing process, then places the residual alcohol that volatilizees at room temperature;
(10) add the distilled water that contains in right amount RNaseA or TE solution to dissolve plasmid DNA, 37 ℃ of digestion ,-20 ℃ are frozen or be directly used in subsequent experimental.
1.6.4 enzyme is cut the evaluation positive transformant
The restriction enzyme at amplification segment two ends carries out the double digestion of transformant to be identified.Double digestion system source can be with reference to TARAKA and NEB website.
1.7 segment splicing
Because gene segment all, on cloning vector pMD-19T, utilizes enzyme to cut the target segment is successively coupled together, finally with at the pAUR135 integrative vector be connected.The results are shown in Figure 1,2.
Embodiment 2: contain the conversion of the industrial saccharomyces cerevisiae of the initial pathways metabolism gene of wood sugar
The brewing yeast cell method for transformation is as follows:
2.1 the preparation of yeast saccharomyces cerevisiae Electroporation-competent cells
(1) yeast-inoculated YPD substratum, 30 ℃, 150rpm cultivates 12-14 hour, the YPD substratum of then transferring new (1% inoculation) incubated overnight;
(2) next day, be placed on ice at least 15min by culturing bottle, allows thalline stop growing.By the 50mL centrifuge tube, ultrapure water, the Sorbitol Solution USP of 1M all is placed on precooling on ice, in low-temperature condition;
(3) centrifugal collection thalline, mix thalline (turn upside down and rock, do not blow and beat with liquid-transfering gun) gently with isopyknic ultrapure water, 3000g, and 5min, 4 ℃ of centrifugal collection thalline, abandon supernatant, repeats this step twice;
(4) clean bacterial sediment 4 times with the 1M Sorbitol Solution USP of 20mL precooling, use for the last time 0.5mL, the resuspended yeast cell of 1M Sorbitol Solution USP, be the yeast competent cell,
2.2 turning method, electricity obtains transformant
(1) competent cell is divided and be filled in 1.5ml EP centrifuge tube, add the 3-5ul linearization plasmid, mix with light finger bomb tube bottom, place on ice;
(2) 40ul competent cell and linearization plasmid mixed solution are added in 0.2 electric revolving cup to ice bath 5-10 minute;
(3) yeast parameter " fungi " is set, " Sc02 ", click Pulse;
(4) take out electric revolving cup, add rapidly the sorbyl alcohol of 1ml ice bath, blow and beat gently with rifle, transfer in 5ml sterilizing centrifuge tube, 30 ℃ standing, hatches 3h;
(5) 3000g is centrifugal 5 minutes;
(6) concentrated, the YPD-Aba that coating contains 2.5ug/ml
+dull and stereotyped.
(7) flat board is cultivated in 30 ℃ of incubators, to yeast transformant, the clone grows.
(8) second take turns screening YPGal flat board, with toothpick, the bacterium colony on first run flat board are chosen to YPGal upper, then cultivate 16 hours for 30 ℃, and the clone that can grow is for being incorporated into the positive colony on the yeast chromosomal genome.
(9) random 12 bacterium colonies of picking are cultivated in the YPD liquid nutrient medium (not containing any microbiotic) again, then carry genomic dna, the integration of three genes of PCR checking.The results are shown in Figure 3.
Embodiment 3: wood sugar recombinant bacterium ZQ5 is in different xylose and glucose concentration bottom fermentation results
Because wild-type S.cerevisiae4126 does not consume any wood sugar, therefore only listed the result of ZQ5 under 50g/l wood sugar and 100g/l table sugar concentration.As shown in Figure 4, ZQ5 can run out of all sugar within 2 weeks.
<110 > Dalian University of Technology
<120 > a strain chromosomal integration is expressed an industrial strain of S.cerevisiae strain of xylose metabolism approach
<141>?2013－07－03
<160>?22
<210>?1
<211>?33
<212>?DNA
<213 > artificial sequence
<400>?1
TTGGCGCGCC?TGCCTTCTAT?TAAGTTGAAC?TCT?33
<210>?2
<211>?35
<212>?DNA
<213 > artificial sequence
<400>?1
TGCTCCGGAT?TATAAGACGA?AGATAGGAAT?CTTGT?35
<210>?3
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
TGGGGCCCCA?ACTCAAG?ACGCACAGA?26
<210>?4
<211>?31
<212>?DNA
<213 > artificial sequence
<400>?1
TTGGCGCGCC?TGTAAGTTTC?ACGAGGTTCT?A?31
<210>?5
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
TGCTCCGGAT?TACATTCACG?CCCTCC?26
<210>?6
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
TTGGCGCG?CGCAAATTAAAG?CCTTCG?26
<210>?7
<211>?27
<212>?DNA
<213 > artificial sequence
<400>?1
TTGGCGCGCC?AACTCAAGAC?GCACAGA?27
<210>?8
<211>?31
<212>?DNA
<213 > artificial sequence
<400>?1
TTGCCTNAGG?TGTAAGTTTC?ACGAGGTTCT?A?31
<210>?9
<211>?31
<212>?DNA
<213 > artificial sequence
<400>?1
TTGCCTNAGG?ATGACTGCTA?ACCCTTCCTT?G?31
<210>?10
<211>?28
<212>?DNA
<213 > artificial sequence
<400>?1
TGGCGGCCGT?TACTCAGGGC?CGTCAATG?28
<210>?11
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
TGGCGGCCGT?TACATTCACG?CCCTCC?26
<210>?12
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
CTAGCTAGCG?CAAATTAAAG?CCTTCG?26
<210>?13
<211>?34
<212>?DNA
<213 > artificial sequence
<400>?1
CTAGCTAGCA?AGAAATGATG?GTAAATGAAA?TAGG?34
<210>?14
<211>?33
<212>?DNA
<213 > artificial sequence
<400>?1
ATAAGAATGC?GGCCGCGC?GCGGAGGGGTGT?TAC?33
<210>?15
<211>?41
<212>?DNA
<213 > artificial sequence
<400>?1
ATAAGAATGC?GGCCGCATGT?TGTGTTCAGT?AATTCAGAGA?C?41
<210>?16
<211>?33
<212>?DNA
<213 > artificial sequence
<400>?1
CCAATGCATT?TAGATGAGAG?TCTTTTCCAG?TTC?33
<210>?17
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
CCAATGCATT?TACATTCACG?CCCTCC?26
<210>?18
<211>?26
<212>?DNA
<213 > artificial sequence
<400>?1
TCCCCGCGCGC?AAATTAAAG?CCTTCG?26
<210>?19
<211>?28
<212>?DNA
<213 > artificial sequence
<400>?1
TCCCCCGGGC?CAGGCGCCTT?TATATCAT?28
<210>?20
<211>?37
<212>?DNA
<213 > artificial sequence
<400>?1
CGGGGTACCT?GGGGCCCTTT?GCGAAACCCT?ATGCTCT?37
<210>?21
<211>?36
<212>?DNA
<213 > artificial sequence
<400>?1
TTCGAGCTCT?CCCGCGCAAT?GGAAGGTCGG?GATGAG?36
<210>?22
<211>?28
<212>?DNA
<213 > artificial sequence
<400>?1
CGGAATTCAT?AAAGCAGCCG?CTACCAAA?28
Claims (3)
1. a strain chromosomal integration is expressed an industrial strain of S.cerevisiae strain (Saccharomyces cerevisiae) of xylose metabolism approach, it is characterized in that: described bacterial strain is the industrial saccharomyces cerevisiae ZQ5 without any selection markers gene, it is registered on the books and is numbered CGMCC7463, be preserved in China Committee for Culture Collection of Microorganisms's common micro-organisms center, preservation date is on April 11st, 2013.
2. chromosomal integration according to claim 1 is expressed an industrial strain of S.cerevisiae strain of xylose metabolism approach, it is characterized in that: described bacterial strain contains three xylose utilization gene Xylose reductase gene PsXR, wood sugar desaturase PsXDH, xylulokinase gene ScXK, the first two gene is from pichia stipitis Pichia stipitis JCM10742, and last gene is from laboratory pattern Saccharomyces Cerevisiae in S 288c.
3. chromosomal integration according to claim 1 is expressed an industrial strain of S.cerevisiae strain of xylose metabolism approach, and it is characterized in that: described bacterial strain contains the xylose metabolism approach, utilizes the wood sugar growth.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310164960.7A CN103484388A (en) | 2013-05-07 | 2013-05-07 | Industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201310164960.7A CN103484388A (en) | 2013-05-07 | 2013-05-07 | Industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN103484388A true CN103484388A (en) | 2014-01-01 |
Family
ID=49824984
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201310164960.7A Pending CN103484388A (en) | 2013-05-07 | 2013-05-07 | Industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN103484388A (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103320333A (en) * | 2013-05-16 | 2013-09-25 | 大连理工大学 | Industrial saccharomycescerevisiae recombination bacterial strain carrying chromosome dispersal integration xylose gene |
| CN104974945A (en) * | 2015-06-26 | 2015-10-14 | 中国石油天然气股份有限公司 | Saccharomyces cerevisiae of overexpression MIG1 genes and preparing method and application of saccharomyces cerevisiae |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101463328B (en) * | 2007-12-19 | 2011-06-29 | 中国科学院微生物研究所 | Metabolic Engineering yeast using xylose fermentation for producing ethanol |
| CN102220254A (en) * | 2010-04-14 | 2011-10-19 | 新疆农业科学院生物质能源研究所 | Recombinant saccharomyces cerevisiae engineering strain and application thereof |
| CN102732437A (en) * | 2012-06-04 | 2012-10-17 | 中国科学院微生物研究所 | Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol |
-
2013
- 2013-05-07 CN CN201310164960.7A patent/CN103484388A/en active Pending
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101463328B (en) * | 2007-12-19 | 2011-06-29 | 中国科学院微生物研究所 | Metabolic Engineering yeast using xylose fermentation for producing ethanol |
| CN102220254A (en) * | 2010-04-14 | 2011-10-19 | 新疆农业科学院生物质能源研究所 | Recombinant saccharomyces cerevisiae engineering strain and application thereof |
| CN102732437A (en) * | 2012-06-04 | 2012-10-17 | 中国科学院微生物研究所 | Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol |
Non-Patent Citations (3)
| Title |
|---|
| ELIASSON ET AL.: "The xylose reductase/xylitol dehydrogenase/xylulokinase ratio affects product formation in recombinant xylose-utilising Saccharomyces cerevisiae", 《ENZYME AND MICROBIAL TECHNOLOGY》 * |
| 王颖等: "木糖代谢工程在酿酒酵母工业菌株中的建立", 《代谢工程和工业生物技术研讨会论文集,2004年》 * |
| 郭亭等: "工业酿酒酵母木糖代谢途径的建立及酒精发酵初步研究", 《甘蔗糖业》 * |
Cited By (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN103320333A (en) * | 2013-05-16 | 2013-09-25 | 大连理工大学 | Industrial saccharomycescerevisiae recombination bacterial strain carrying chromosome dispersal integration xylose gene |
| CN104974945A (en) * | 2015-06-26 | 2015-10-14 | 中国石油天然气股份有限公司 | Saccharomyces cerevisiae of overexpression MIG1 genes and preparing method and application of saccharomyces cerevisiae |
| CN104974945B (en) * | 2015-06-26 | 2018-05-04 | 中国石油天然气股份有限公司 | A kind of saccharomyces cerevisiae of overexpression MIG1 genes and preparation method and application |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| ES2651076T3 (en) | Suitable cell for the fermentation of a mixed sugar composition | |
| ES2256742T5 (en) | A modified yeast that consumes L-arabinose | |
| CN105199976B (en) | Recombinant saccharomyces cerevisiae strain for co-fermenting glucose and xylose and application thereof | |
| CN103849576B (en) | One strain has the recombinant Saccharomyces cerevisiae bacterial strain of stress tolerance | |
| CN104718290A (en) | Xylose isomerases and their uses | |
| WO2015100856A1 (en) | Genetic recombinant saccharomyces cerevisiae capable of degrading and utilizing kitchen wastes | |
| ES2755680T3 (en) | Yeast cell consuming acetate | |
| CN104164375A (en) | Construction and application of high-temperature high-yield engineering strain for producing ethanol | |
| CN103820347B (en) | One strain has an industrial strain of S.cerevisiae strain of Acetate tolerance | |
| CN101463328B (en) | Metabolic Engineering yeast using xylose fermentation for producing ethanol | |
| CN105368731B (en) | Saccharomyces cerevisiae strain for expressing xylose isomerase and construction method thereof | |
| CN105647822A (en) | Restructured mortierella alpina with overexpression from omega-3 desaturase of phytophthora parasitica, and establishment method and application thereof | |
| CN103374542A (en) | Method for increasing consumption rate of clostridium beijerinckii xylose | |
| CN105567579A (en) | Recombinant strain for expressing heterogenous Omega-3 desaturase in mortierella alpina and construction method thereof | |
| CN111088177B (en) | Construction and application of heat-resistant yeast engineering bacteria for producing glycerol under high-temperature aerobic condition | |
| CN101878308B (en) | A process for the preparation of ethanol from starch | |
| CN103484388A (en) | Industrial saccharomyces cerevisiae bacterial strain realizing chromosome integrative expression of xylose metabolic pathways | |
| CN107937296A (en) | One kind has acetic acid furfural vanillic aldehyde tolerance recombinant Saccharomyces cerevisiae and preparation method, application | |
| CN103131720A (en) | Fungi xylose isomerase gene and application thereof | |
| CN104974945B (en) | A kind of saccharomyces cerevisiae of overexpression MIG1 genes and preparation method and application | |
| CN103320333B (en) | Industrial saccharomycescerevisiae recombination bacterial strain carrying chromosome dispersal integration xylose gene | |
| CN102604850B (en) | Brewing yeast strain capable of metabolizing xylose | |
| CN102732437B (en) | Saccharomyces cerevisiae engineering bacterium and its application in production of ethanol | |
| CN106929530A (en) | A kind of method for improving yeast cells to composite inhibitor tolerance | |
| WO2017037745A1 (en) | An integrated process for production of carbohydratases, ethanol, and xylitol using an isolated candida strain |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C05 | Deemed withdrawal (patent law before 1993) | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20140101 |