CN102888420B - Gene for coding xylose isomerase and application thereof - Google Patents
Gene for coding xylose isomerase and application thereof Download PDFInfo
- Publication number
- CN102888420B CN102888420B CN201210387377.8A CN201210387377A CN102888420B CN 102888420 B CN102888420 B CN 102888420B CN 201210387377 A CN201210387377 A CN 201210387377A CN 102888420 B CN102888420 B CN 102888420B
- Authority
- CN
- China
- Prior art keywords
- gene
- xylose isomerase
- pgi
- application
- isomerase
- 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.)
- Expired - Fee Related
Links
Images
Classifications
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02P—CLIMATE CHANGE MITIGATION TECHNOLOGIES IN THE PRODUCTION OR PROCESSING OF GOODS
- Y02P20/00—Technologies relating to chemical industry
- Y02P20/50—Improvements relating to the production of bulk chemicals
- Y02P20/52—Improvements relating to the production of bulk chemicals using catalysts, e.g. selective catalysts
Landscapes
- Preparation Of Compounds By Using Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Enzymes And Modification Thereof (AREA)
Abstract
The invention relates to a gene Pgi for coding xylose isomerase, belonging to the technical field of gene engineering. The gene contains a nucleotide sequence shown as SEQ ID NO.1, a protein coded by the gene contains an amino acid sequence shown as SEQ ID NO.2, and the protein is used for producing xyloketose from xylose used as the raw material or producing levulose from glucose used as the raw material. The invention can widen the source and application of the xylose isomerase gene and better serve the industrial production of xyloketose or levulose.
Description
Technical field
The present invention relates to gene engineering technology field, specifically a kind of gene of the xylose isomerase of encoding and take application or the application of glucose as raw material direct production fructose of take that wood sugar is raw material direct production xylulose by the protein of the genes encoding of this coding xylose isomerase.
Background technology
Xylose isomerase (xylose isomerase, XI) (EC5.3.1.5) is called again glucose isomerase (glucose isomerase, GI), is a kind of industrial enzymes with significant application value.It can turn to xylulose by xylose isomerase, in the process of utilizing lignocellulose production of ethanol from microbial fermentation, plays keying action; Also can form fructose by transforming glucose, industrial for the production of high fructose syrup (high fructose corn syrup, HFCS) or the important zymin (Jensen of high fructose syrup or isomery syrup, V.J.and Rugh, S.1987.Industrial scale application of immobilized glucose isomerase.Methods Enzymol.136,356-370.).
Xylose isomerase is the key enzyme of pentose-phosphate pathway.Hemicellulose accounts for 50% of total amount in lignum, and xylose isomerase endonuclease capable is converted into xylulose by wood sugar, is xylan is converted into indispensable enzyme in ethanol approach.According to relevant data, utilize Production of Alcohol from Lignocellulose will reduce by 20% (the Dawid Brat nearly that prior art is produced cost that ethanol is spent, Eckhard Boles, and Beate Wiedemann, 2009, Functional Expression of a Bacterial Xylose Isomerase in Saccharomyces cerevisiae, Appl.Environ.Microbiol.75 (8): 2304-2311.).Therefore, research and development xylose isomerase is the another research approach of exploitation bioenergy for industrial production alcohol
kaisa Karhumaa, Marie Jeppsson, et al.2007.Metabolic Engineering for Pentose Utilization in Saccharomyces cerevisiae, Advances in Biochemical Engineering/Biotechnology, 108,147-177.).
Xylose isomerase also can be applicable to glucose isomerase and turns in the food industries of fructose.Supply falls short of demand for sugar (comprising sucrose, beet sugar, high fructose syrup) in the world, and consumption is greater than turnout, and on world market, sugared valency goes up.In addition, high fructose syrup is widely used as the sweeting agent of beverage, jelly, candy, ice cream and seasoning food, also can replace honey to be used for pharmaceutical industry, or for producing N.F,USP MANNITOL etc. on chemical industry, compare with sucrose, price is slightly low, sugariness is suitable, therefore the market requirement of high fructose syrup is in continuous increase, and naturally, research glucose isomerase is significant.
As far back as nineteen fifty-seven, have a liking in water pseudomonas and find that GI is active, find that there is afterwards nearly hundred kinds of bacteriums and actinomycetes and produced GI, and be mostly to produce intracellular enzyme, only a few bacterial strain produces extracellular enzyme (Zhu Guoping, Cheng Yang, Gong Chunhong etc., the research progress of bioengineering of 2000. glucose isomerases, Progress in Biochemistry and Biophysics, 27 (2): 127-130.).The seventies China's glucose isomerase that begins one's study, occurred that colibacillary colt Glucose Isomerase Gene clones and express in streptomycete, also there is Yeast engineering bacteria (Yi Guohui, Yin Huixiang, Zhang Ailian, 2011, with the pAOX1 expression system of P.pastoris, express xylose isomerase, biotechnology, 21(1): 17-19.) and colibacillus engineering (fourth jasmine, Xu Wei, strict and impartial etc., 2006, Thermus thermophilus HB8 glucose isomerase is at expression in escherichia coli, biological processing, 4(2): 42-45.), but studying at present more is that the colt Glucose Isomerase Gene containing in intestinal bacteria is expressed in the yeast of allos or streptomycete, a few studies is also devoted to extract xylose isomerase gene producing and ethanol (Yoshiaki Umemoto from marine algae, Toshiyuki Shibata, Toshiyoshi Araki.2012, D-Xylose Isomerase from a Marine Bacterium, Vibrio sp.Strain XY-214, and D-Xylulose Production from β-1, 3-Xylan.Mar Biotechnol.14:10 – 20), the research also having is in intestinal bacteria, to build recombinant bacterium and express to have organized enzyme, and study its zymologic property, and carry out suitable transgenation, to build, be applicable to the mutant that high fructose syrup is produced.
The biological property of research glucose isomerase or xylose isomerase finds, this enzyme heat stability is high, generally all at 60 ℃, even reaches above 90 ℃, and space structure is relevant with special active conserved sequence closely with it for this characteristic; Its substrate, except glucose and xylose, also has D-ribose, L-arabinose, L-rhamnosyl, D-allose and deoxyglucose; Various divalent-metal ions also have and promote or restraining effect its enzyme work.A lot of research is all devoted to transgenation and is reached its biological characteristics of improvement, to obtaining the result that has academic and commercial value.
Take xylose isomerase as keyword lookup State Intellectual Property Office patent retrieval database, there are altogether 33 patents of invention, have 10 that relate to xylose isomerase gene and application thereof, wherein there are 3 to be respectively about guanosine diphosphate(GDP) xylose isomerase, the gene of the xylose isomerase of japonica rice and the xylose isomerase of barley and application thereof, 2 is about xylose isomerase enzyme mutant and application thereof, have 5 to be xylose isomerase gene and application thereof about different sources--respectively from Arabidopis thaliana, the grand genomic library of bovine rumen liquid, the total RNA of people's tire brain, sorangium cellulosum (Sorangium cellulosum) 157-2, clostridium (Clostridium) and fusobacterium (Fusobacterium) belong to the tunicate (tunicate) of bacterium or Ciona (Ciona) genus, other patent is that various bacterial strains about containing xylose isomerase and xylose isomerase are in the application without in field.
Take glucose isomerase as keyword lookup State Intellectual Property Office patent retrieval database, occur altogether 42 patents of invention, only have 1 to be about warm colt Glucose Isomerase Gene and application thereof; 12 patents of invention are to describe to utilize engineered method to build glucose isomerase mutant, and have described their characteristic and application; Other patent of invention is to describe the application of immobilized glucose isomerase and the application in producing the aspects such as alcohol, high fructose syrup, N.F,USP MANNITOL of glucose isomerase.
Summary of the invention
The gene that the object of this invention is to provide a kind of xylose isomerase of encoding, and the protein of the coded by said gene by this coding xylose isomerase is used for take wood sugar as raw material production xylulose or take glucose as raw material production fructose, thereby realize source and the application thereof of widening xylose isomerase gene.
The present invention realizes the technical scheme that above-mentioned purpose takes: a kind of gene Pgi of the xylose isomerase of encoding, its nucleotide sequence is as shown in sequence table SEQ ID NO.1.
The gene Pgi of coding xylose isomerase of the present invention is that separation obtains from the Ke Shi series bacillus of laboratory screening.The conserved regions of xylose isomerase gene sequence, the foreign DNA of base sequence are by 768 based compositions, the open reading frame that contains complete xylose isomerase gene Pgi (Open Reading Frame, ORF), the initiation codon of Pgi gene is ATG, and terminator codon is TAA.
By the coded protein of gene Pgi of described coding xylose isomerase, its aminoacid sequence is as shown in SEQ ID NO.2.
Of the present invention by the coded protein of gene Pgi, the xylose isomerase product P gi of gene Pgi coding, by 255 amino acid, formed, with Pgi catalysis territory homology the highest be the catalysis territory (e-value is 9e-111) of the sugar phosphate isomerase/epimerase of Paenibacillus polymyxa Paenibacillus polymyxa E681, both similarity=159/255 (62%), homogeny=203/255 (80%).
The invention still further relates to the expression vector of the gene Pgi that contains coding xylose isomerase of the present invention, and utilize this expression vector to transform the host cell obtaining.
Of the present invention by the coded protein of gene Pgi for take wood sugar as raw material production xylulose or take glucose as raw material production fructose.
The gene Pgi of the present invention's separated xylose isomerase that obtains encoding from Ke Shi series bacillus, source and the application thereof that can widen xylose isomerase gene, serve the industrial production of xylulose or fructose better.
Accompanying drawing explanation
Fig. 1 is the SDS-PAGE figure of xylose isomerase Pgi purified.
From Fig. 1, recognize, the molecular weight of xylose isomerase Pgi purified is 29kDa.
Embodiment
Below in conjunction with embodiment, technical scheme of the present invention is described further:
Material used comprises in an embodiment of the present invention: intestinal bacteria (Escherichia coli) strain XL1-Blue, carrier pMD19-T(are purchased from Dalian TaKaRa company); Expression vector pQE30, intestinal bacteria strain M15(are purchased from Novagen company); The reagent such as restriction enzyme, modifying enzyme.
(1) structure of Paenibacillus cookii GX-4 genomic library
The genomic dna of P.cookii GX-4 is the bacterial genomes DNA extraction test kit Biospin Bacteria Genomic DNA Extraction Kit(catalog number (Cat.No.) BSC12S1 according to BioFlux company) operation instruction extract.
The genomic library construction of P.cookii GX-4 is to prepare test kit CopyControl according to the library of Epicentre company
tMfosmid Library Production Kit(catalog number (Cat.No.) CCFOS110) working instructions carry out.The genomic library of the P.cookii GX-4 building is applied to containing the LA of paraxin (12.5 μ g/mL), dull and stereotyped ([every 100ml contains LA substratum: peptone: 1g, yeast powder: 0.5g, NaCl:0.5g, agar powder: 1.5g).Result obtains approximately 4231 transformants altogether.
(2) mensuration of xylose isomerase gene Pgi sequence
At random from containing transformant of picking the LA flat board of paraxin (12.5 μ g/mL), be inoculated into LB liquid nutrient medium (substratum [and every 100ml contains: peptone: 1g, yeast powder: 0.5g, NaCl:0.5g) in, 37 ℃ of concussions are cultivated 12 hours.Extract the plasmid DNA of this transformant, after cutting with restriction enzyme BamHI enzyme, the DNA fragmentation of acquisition is connected with the dephosphorylized pUC19 plasmid of cutting through BamHI enzyme.Again connection product is transformed into in E.colistrain XL1 blue, (method for transformation is CaCl
2method), containing on the LA flat board of penbritin (100 μ g/mL), screening subclone transformant.Result obtains approximately 312 subclone transformants altogether.Further extract the wherein plasmid DNA of 6 subclone transformants, with after restriction enzyme BamHI complete degestion, carry out 0.9% agarose gel electrophoresis analysis, these 6 subclone transformants of result can be cut outside the carrier segments of next 2.7kb by enzyme, and one of them subclone can also provide the DNA fragmentation that a size is 9.4kb.So, this subclone is checked order, and called after D3-9.D3-9 delivers Hua Da genome company and measures DNA nucleotide sequence.
(3) nucleotide sequence analysis of xylose isomerase gene Pgi
With NCBI(National Center for Biotechnology Information) on software DNA sequence dna is analyzed as ORF finder, Blast.The open reading frame of gene Pgi (Open Reading Frame, ORF) is comprised of 768 Nucleotide, and sequence is as SEQ ID NO:1.Wherein, the initiator codon of Pgi gene is ATG, and terminator codon is TAA.
(4) amino acid sequence analysis of the product P gi of xylose isomerase gene Pgi coding
One of xylose isomerase gene Pgi coding contains 255 amino acid whose protein, by the theoretical molecular size of this protein of DNAStar software prediction, is 29028.74 dalton.
With simple assemblies structural research instrument (Simple Modular Architecture Research Tool, SMART, http://smart.embl-heidelberg.de) analyze the unit construction of xylose isomerase Pgi, result is to be xylose isomerase (AP_endonuc_2) functional domain from the 23-204 position of N end.
(5) clone of xylose isomerase gene Pgi and expression
Use upstream primer 5 '-CGTGGATCCTTGAAGCTGGGCTTGCAACTTTATAC-3 ' and downstream primer 5 '-GTGAAGCTTGACATGAACAGGTCCTCCATTTTTTG-3 ', by polymerase chain reaction (PCR) amplification xylose isomerase gene Pgi, with restriction enzyme BamH I and Hind III enzyme, cut after xylose isomerase gene Pgi, be connected with the expression vector pQE30 cutting with Hind III enzyme through BamH I.Again connection product is transformed into in E.colistrain XL1 blue, (method for transformation is CaCl
2method), be applied on the LA flat board containing penbritin (100 μ g/mL), flat board be placed in to 37 ℃ of thermostat containers and cultivate 14 hours.Then further extract the plasmid DNA of the clone on flat board, and by its called after pQE-Pgi, with restriction enzyme BamH I and Hind III enzyme, cut after pQE-Pgi, carry out 0.9% agarose gel electrophoresis analysis, outside the carrier DNA fragment of result pQE-Pgi apart from a 3.4kb, also have a size to be about the exogenous dna fragment of 768bp.
To contain plasmid pQE-Pgi recombination bacillus coli M15 inoculation and contain in the LB substratum of penbritin (100 μ g/mL) to 600mL, 37 ℃ of shaking culture, are 0.4 o'clock until OD600, and adding IPTG to make its final concentration is 0.5mmol/L, induce 20 hours for 20 ℃.The centrifugal 3min of 11000rpm, collects thalline, with the resuspended thalline of phosphoric acid buffer of 4mL pH7.0100mmol/L, and the broken born of the same parents of ultrasonic wave 9 minutes.The centrifugal 20min of 12000rpm, gets supernatant and carries out protein purification below.The nickel affinity chromatography colloid that adds 1mL50% by every 4mL supernatant liquor, turns and shakes 60 minutes with 200 at 4 ℃, and mixture is filled into pillar, collects effluent.Add 1mL dcq buffer liquid (50mmol/LNaH
2pO4,300mmol/LNaCl, 20mmol/L imidazoles, pH8.0) in pillar, slowly stirs, and collects effluent.Repeat rinse step 4 times.Add elution buffer (50mmol/LNaH
2pO4,300mmol/LNaCl, 250mmol/L imidazoles, pH8.0) elute protein.Collect the protein soln of wash-out, with polyacrylamide gel electrophoresis (SDS-PAGE) checking of sex change, find that there is the protein band of object size.
(6) determination of activity of xylose isomerase Pgi
Pgi be take wood sugar and is adopted halfcystine-carbazole method as substrate conversion becomes the enzyme activity determination of xylulose: the xylose isomerase Pgi enzyme liquid, the 50 μ l10mM MgCl that get 3 μ l purifying
2solution, 50 μ l0.1mol/L D-xylose solutions (being dissolved in the phosphoric acid buffer of pH7.0,100mM) mix, 80 ℃ of effects after 15 minutes, and boiling water bath termination reaction.Get the H that 100 μ l enzyme reaction solutions add 6mL13mol/L immediately
2sO
4, 200 μ l1.5% cysteine hydrochlorides and 200 μ l0.12% carbazole spirituous solutions (use anhydrous alcohol solution carbazole, place in brown bottle and use after 24 hours) mix, 25 ℃ of reactions 30 minutes.
With spectrophotometric instrumentation absorbancy OD
540.The xylulose absorbancy canonical plotting of absorbance measurement value and different content is made comparisons and is calculated enzyme work.
In standard reaction mixture, the enzyme activity unit of xylose isomerase (U) is defined as per minute catalysis and produces the required enzyme amount of 1 μ moL xylulose.The enzyme activity of Pgi conversion wood sugar generation xylulose reaches and is up to 21.789U/mg.
Pgi be take glucose and is generated the enzyme activity determination of fructose as substrate: get the xylose isomerase Pgi enzyme liquid, 50 μ l3mol/L glucose solutions, 400 μ l100mM pH7.0 Potassium Hydrogen Phthalate-imidazole buffers of 3 μ l purifying (containing 5mm/L Mg
2+, 1mm/L Co
2+) mix, 80 ℃ of effects after 15 minutes, by 500 μ l50% trichoroacetic acid(TCA) stopped reactions.Get the H that 100 μ l reaction solutions add 6mL13mol/L immediately
2sO
4, 200 μ l1.5% cysteine hydrochlorides and 200 μ l0.12% carbazole spirituous solutions (use anhydrous alcohol solution carbazole, place in brown bottle and use after 24 hours) mix, 25 ℃ of reactions 30 minutes.
With spectrophotometric instrumentation absorbancy OD
540.The fructose absorbancy canonical plotting of absorbance measurement value and different content is made comparisons and is calculated enzyme work.
In above-mentioned reaction mixture, the enzyme activity unit of xylose isomerase (U) is defined as per minute catalysis and produces the required enzyme amount of 1 μ moL fructose.The enzyme activity of Pgi transforming glucose generation fructose reaches and is up to 1356U/mg.
Claims (5)
1. the encode gene Pgi of xylose isomerase, is characterized in that, its nucleotide sequence is as shown in sequence table SEQ ID NO.1.
2. by the coded protein of gene Pgi claimed in claim 1, it is characterized in that, its aminoacid sequence is as shown in SEQ ID NO.2.
3. an expression vector, is characterized in that, contains gene Pgi claimed in claim 1.
4. a host cell, is characterized in that, contains prokaryotic cell prokaryocyte or eukaryotic cell that expression vector claimed in claim 3 transforms.
5. protein claimed in claim 2 take application or the application of glucose as raw material production fructose of take that wood sugar is raw material production xylulose.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210387377.8A CN102888420B (en) | 2012-10-12 | 2012-10-12 | Gene for coding xylose isomerase and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201210387377.8A CN102888420B (en) | 2012-10-12 | 2012-10-12 | Gene for coding xylose isomerase and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN102888420A CN102888420A (en) | 2013-01-23 |
| CN102888420B true CN102888420B (en) | 2014-04-09 |
Family
ID=47532102
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201210387377.8A Expired - Fee Related CN102888420B (en) | 2012-10-12 | 2012-10-12 | Gene for coding xylose isomerase and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN102888420B (en) |
Families Citing this family (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| JP6977231B2 (en) | 2015-07-13 | 2021-12-08 | マラ リニューアブルズ コーポレーション | Enhancement of metabolism of C5 organic carbon by microorganisms |
Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101283096A (en) * | 2005-09-06 | 2008-10-08 | 卡吉尔公司 | Thermostable xylose isomerase enzymes |
| CN101323858A (en) * | 2008-07-24 | 2008-12-17 | 河南天冠企业集团有限公司 | Xylose isomerase, and encoding gene and use thereof |
| CN102643844A (en) * | 2011-02-21 | 2012-08-22 | 中国农业科学院作物科学研究所 | Function and application of xylose isomerase gene of non-glutinous rice |
-
2012
- 2012-10-12 CN CN201210387377.8A patent/CN102888420B/en not_active Expired - Fee Related
Patent Citations (3)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN101283096A (en) * | 2005-09-06 | 2008-10-08 | 卡吉尔公司 | Thermostable xylose isomerase enzymes |
| CN101323858A (en) * | 2008-07-24 | 2008-12-17 | 河南天冠企业集团有限公司 | Xylose isomerase, and encoding gene and use thereof |
| CN102643844A (en) * | 2011-02-21 | 2012-08-22 | 中国农业科学院作物科学研究所 | Function and application of xylose isomerase gene of non-glutinous rice |
Non-Patent Citations (1)
| Title |
|---|
| Kim, J.F.，等.sugar phosphate isomerase/epimerase [Paenibacillus polymyxa E681].《Genbank登录号YP_003871247》.2012,全文. * |
Also Published As
| Publication number | Publication date |
|---|---|
| CN102888420A (en) | 2013-01-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Inoue et al. | Discovery of a novel alginate lyase from Nitratiruptor sp. SB155-2 thriving at deep-sea hydrothermal vents and identification of the residues responsible for its heat stability | |
| Ji et al. | The molecular basis of endolytic activity of a multidomain alginate lyase from Defluviitalea phaphyphila, a representative of a new lyase family, PL39 | |
| Xu et al. | Structural and molecular basis for the substrate positioning mechanism of a new PL7 subfamily alginate lyase from the arctic | |
| JP5468680B2 (en) | Novel alpha-neo agarobiose hydrolase and method for obtaining monosaccharides using the same | |
| Peng et al. | A novel bifunctional endolytic alginate lyase with variable alginate-degrading modes and versatile monosaccharide-producing properties | |
| JP2020506704A (en) | Engineered glycosyltransferase and steviol glycoside glycosylation methods | |
| Groisillier et al. | The mannitol utilization system of the marine bacterium Zobellia galactanivorans | |
| CN106414728A (en) | Agarooligosaccharide hydrolase and method for producing 3,6-anhydro-l-galactose and galactose from agarose by using same | |
| Mine et al. | Expression from engineered Escherichia coli chromosome and crystallographic study of archaeal N, N′‐diacetylchitobiose deacetylase | |
| Bhattacharya et al. | Review of the enzymatic machinery of Halothermothrix orenii with special reference to industrial applications | |
| Rozeboom et al. | Crystal structure of α-1, 4-glucan lyase, a unique glycoside hydrolase family member with a novel catalytic mechanism | |
| Li et al. | Biochemical characterization of a thermophilic hyaluronate lyase TcHly8C from Thermasporomyces composti DSM22891 | |
| CN103114100B (en) | Beta-glucosaccharase gene S-bgl3 and application thereof | |
| CN102888420B (en) | Gene for coding xylose isomerase and application thereof | |
| CN112899257B (en) | Thermophilic chitinase Chi304 mutant and application thereof in degradation of chitin | |
| CN102888416A (en) | Beta-glycosidase gene for coding glycosyl hydrolase family and application thereof | |
| CN106701844A (en) | Method using klebsiella pneumoniae to produce xylonic acid | |
| KR20100040438A (en) | A novel agarase and an enzymatic production method of agarooligosaccharide from agarose using the same | |
| CN106566824B (en) | A kind of glucose isomerase, gene, carrier, engineering bacteria and its application | |
| Chen et al. | Enhanced the catalytic efficiency and thermostability of maltooligosyltrehalose synthase from Arthrobacter ramosus by directed evolution | |
| CN104087604A (en) | Genetic expression sequence of inulin fructotransferase | |
| Ren et al. | Structure and function of aldopentose catabolism enzymes involved in oxidative non-phosphorylative pathways | |
| Fatima et al. | Xylose isomerases from thermotogales. | |
| Nirwantono et al. | Exploring an l-arabinose isomerase from cryophile bacteria Arthrobacter psychrolactophilus B7 for d-tagatose production | |
| Kluskens et al. | Molecular characterization of the glucose isomerase from the thermophilic bacterium Fervidobacterium gondwanense |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| C06 | Publication | ||
| PB01 | Publication | ||
| C10 | Entry into substantive examination | ||
| SE01 | Entry into force of request for substantive examination | ||
| C14 | Grant of patent or utility model | ||
| GR01 | Patent grant | ||
| CF01 | Termination of patent right due to non-payment of annual fee |
Granted publication date: 20140409 Termination date: 20161012 |
|
| CF01 | Termination of patent right due to non-payment of annual fee |