WO2010090359A1 - Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase - Google Patents

Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase Download PDF

Info

Publication number
WO2010090359A1
WO2010090359A1 PCT/KR2009/000544 KR2009000544W WO2010090359A1 WO 2010090359 A1 WO2010090359 A1 WO 2010090359A1 KR 2009000544 W KR2009000544 W KR 2009000544W WO 2010090359 A1 WO2010090359 A1 WO 2010090359A1
Authority
WO
WIPO (PCT)
Prior art keywords
arabinitol
dehydrogenase
arabinitol dehydrogenase
ribulose
gene
Prior art date
Application number
PCT/KR2009/000544
Other languages
English (en)
Korean (ko)
Inventor
이정걸
문희정
마니시티와리
Original Assignee
건국대학교 산학협력단
Priority date (The priority date 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 date listed.)
Filing date
Publication date
Application filed by 건국대학교 산학협력단 filed Critical 건국대학교 산학협력단
Priority to PCT/KR2009/000544 priority Critical patent/WO2010090359A1/fr
Publication of WO2010090359A1 publication Critical patent/WO2010090359A1/fr

Links

Images

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12P—FERMENTATION OR ENZYME-USING PROCESSES TO SYNTHESISE A DESIRED CHEMICAL COMPOUND OR COMPOSITION OR TO SEPARATE OPTICAL ISOMERS FROM A RACEMIC MIXTURE
    • C12P19/00—Preparation of compounds containing saccharide radicals
    • C12P19/02—Monosaccharides

Definitions

  • the present invention relates to a novel L-arabinitol dehydrogenase and a method for preparing L-ribulose using the same, and more particularly, to L-arabinitol dehydrogenase, a nucleic acid molecule encoding the same, and the nucleic acid molecule. It relates to a vector, a transformant comprising the vector and a method for producing L-ribulose using the L- arabinitol dehydrogenase.
  • the present invention is derived from a study carried out as part of the microbial genome utilization technology development project of the Ministry of Education, Science and Technology [Task No. 2007-A002-0065, Task name: Development of customized oxidoreductase by in vitro coevolution].
  • L-Ribose is the starting material for the synthesis of drugs per many L-form nucleic acids.
  • L-Ribose has been produced mainly by chemical synthesis from L-arabinose, L-xylose, D-glucose, D-galactose, D-ribose or D-mannono-1,4-lactone.
  • chemical synthesis has disadvantages such as low total yield, many chemical reaction steps, complex purification process and by-product formation.
  • biological L-ribose preparation from ribitol or L-ribulose has been studied.
  • L-ribose is an important key pentose sugar that constitutes the backbone in the synthesis of L-ribonucleosides, L-oligoribonucleosides and many other therapeutic agents.
  • L-nucleoside is a high potential candidate that can be used as a therapeutic material because it has high stability against attack of nucleases and the like in the body as compared to D-nucleoside.
  • L-ribose is known to be highly useful as a raw material for manufacturing medicines such as antiviral drugs and anticancer drugs.
  • the present invention solves the above problems, and the first object of the present invention is to provide a gene of L-arabinitol dehydrogenase.
  • a third object of the present invention is to provide a recombinant expression vector containing the gene of the L- arabinitol dehydrogenase.
  • a fifth object of the present invention is to provide a recombinant L-arabinitol dehydrogenase using transformed recombinant E. coli.
  • a sixth object of the present invention is to provide a method for preparing L-ribulose from L-arabinitol using the enzyme.
  • the present invention is characterized by using the L- arabinitol dehydrogenase of the Debaromyces hansenii strain in the production method of L-ribulose.
  • the present invention is characterized by cloning the L- arabinitol dehydrogenase gene from devaromysis Hanseni through Southern hybridization and colony hybridization.
  • the present invention provides an L-arabinitol dehydrogenase having an amino acid sequence of SEQ ID NO: 4 or a functional fragment thereof.
  • the L-arabinitol dehydrogenase is preferably derived from debaromysis hanseni, but is not limited thereto.
  • the arabinitol dehydrogenase is characterized in that it is specific to L- arabinitol.
  • the molecular weight of the enzyme of the present invention is characterized in that 38 kDa.
  • the enzyme of the present invention is characterized by an increase in activity in the presence of 1 mM Mg 2+ or 1 mM NAD + .
  • the present invention also provides a L- arabinitol dehydrogenase gene encoding the enzyme of the present invention.
  • the gene of the present invention preferably has a nucleotide sequence of SEQ ID NO: 3, at least 85%, preferably at least 90% or more, more preferably from the sequence of SEQ ID NO: 3 in consideration of degeneracy of the genetic code, etc. Is preferably a sequence having at least 95% homology, but is not limited thereto.
  • the present invention provides a method for producing L- arabinitol dehydrogenase by culturing a strain transformed with a recombinant expression vector comprising the L- arabinitol dehydrogenase gene of the present invention.
  • the present invention also provides a method for preparing L-ribulose from L-arabitol using the L-aravinitol dehydrogenase of the present invention.
  • L-arabinitol dehydrogenase of the present invention is characterized by having an amino acid sequence represented by SEQ ID NO: 4.
  • at least one amino acid sequence of SEQ ID NO: 4 may be deleted, substituted, or added to one or more amino acids within a range in which the L-arabinitol dehydrogenase activity indicated by the protein having these amino acid sequences is not impaired.
  • Mutant L-arabinitol dehydrogenase into which a mutation of is introduced is also included in the L-arabinitol dehydrogenase according to the present invention.
  • the present invention includes an L-arabinitol dehydrogenase gene encoding an L-arabinitol dehydrogenase having an amino acid sequence of SEQ ID NO: 4, and the gene sequence represented by SEQ ID NO: 3 may be mentioned. have.
  • the L-arabinitol dehydrogenase gene encoding the above-mentioned variant L-arabinitol dehydrogenase obtained by mutating the nucleotide sequences of SEQ ID NO: 3 is also L-arabinitol dehydrogenation according to the present invention. It is included in the enzyme gene.
  • the present invention includes a recombinant vector containing the L-arabinitol dehydrogenase gene and a transformant transformed by the recombinant vector.
  • the present invention also includes a method for producing L-arabinitol dehydrogenase, wherein the transformant is cultured to separate L-arabinitol dehydrogenase from the culture obtained.
  • L-arabinitol dehydrogenase gene of the present invention is isolated from the cells of devaromysis Hanseni. First, chromosomal DNA is obtained from a strain having a L-arabinitol dehydrogenase gene.
  • PCR polymerase chain reaction
  • PCR amplification fragments are labeled with appropriate reagents, and colony hybridization is performed on the chromosomal DNA library to select L-arabinitol dehydrogenase genes (Current Protocols in Molecular Biology, Vol. 1, 603). Page, 1994).
  • DNA fragments containing the L-arabinitol dehydrogenase gene were recovered by recovering the plasmid from the Escherichia coli selected by the above method using alkaline protocols (Current Protocols in Molecular Biology, Vol. 1, p. 161, 1994). You can get it. After determining the nucleotide sequence by the above method, it is possible to obtain the entire gene of the present invention by hybridizing the DNA fragment prepared by digestion by restriction enzymes of the DNA fragment having the nucleotide sequence as a probe.
  • SEQ ID NO: 3 shows the nucleotide sequence of the L- arabinitol dehydrogenase gene of the present invention
  • SEQ ID NO: 4 shows the amino acid sequence encoded by the gene.
  • the transformed microorganism of the present invention is obtained by introducing the recombinant vector of the present invention into a host suitable for the expression vector used when producing the recombinant vector.
  • a host suitable for the expression vector used when producing the recombinant vector for example, when a bacterium such as E. coli is used as a host, the recombinant vector according to the present invention is capable of autonomous replication in the host, and at the same time, a DNA containing a promoter, an L-arabinitol dehydrogenase gene, and It is preferred to have a configuration necessary for the expression of the transcription termination sequence.
  • PGEX-KG was used as the expression vector used in the present invention, but any expression vector satisfying the above requirements can be used.
  • L-arabinitol dehydrogenase In the production of L-arabinitol dehydrogenase according to the present invention, a transformant obtained by transforming a host with a recombinant vector having a gene encoding the same is cultured, and the gene is cultured (cultured cell or culture supernatant). This is done by producing and accumulating the product L-arabinitol dehydrogenase and obtaining the enzyme from the culture.
  • L-arabinitol dehydrogenase Acquisition and purification of L-arabinitol dehydrogenase is performed by centrifuging the cells or supernatant from the cultures obtained, and by cell alone, affinity chromatography, cation or anion exchange chromatography, or the like. I can do it.
  • the present inventors cloned the gene of L-arabinitol dehydrogenase from Devaromysis hanseni to develop an enzyme capable of producing L-ribulose with high yield. It was confirmed that the recombinant strain incorporating the above-described gene can not only prepare L-ribulose from L-arabitol in high yield, but also greatly reduce the production of by-products, and completed the present invention.
  • the present invention is to clone the gene encoding L- arabinitol dehydrogenase from the gene of devaromysis Hanseni in order to produce industrially useful L- arabinitol dehydrogenase, and from the base sequence of the gene Analyze the inferred amino acid sequence.
  • the L-arabinitol dehydrogenase of the present invention is an enzyme which forms L-ribulose by catalyzing the dehydrogenation reaction using L-arabitol as a substrate, and more preferably has L-arabi specificity for dehydrogenation. L-arabinitol dehydrogenase with the ability to convert toll to L-ribulose.
  • L-arabinitol dehydrogenase of the present invention has the following characteristics: (iii) a molecular weight of about 38 kDa; (Ii) enzymatic activity in the presence of NAD + and Mg 2+ ,
  • L-arabinitol dehydrogenase of the present invention exhibits enzymatic activity in the presence of 1 mM NAD + and 1 mM Mg 2+ .
  • the known L-aravinitol dehydrogenase shows low L-ribulose conversion.
  • the L-arabinitol dehydrogenase of the present invention produces L-ribulose with high yield using L-arabinitol. Therefore, the enzyme of the present invention which produces L-ribulose in high yield in L-arabitol will be very specific and will be usefully applied in the production of L-ribulose from sugar mixtures.
  • FIG. 1 is a diagram illustrating a search through Southern hybridization of a fragment having L-arabinitol dehydrogenase in the devaromysis hanseni chromosome (genomic DNA).
  • No restriction enzyme treatment was performed on the mysis hanseni chromosome (genomic DNA), lanes 2, 3, 4, 5, and 6 were respectively treated with restriction enzymes EcoRI, SalI, BamHI, HindIII, and XbaI.
  • Figure 2 is a vector map of the vector pUC-LAD to find a fragment containing the L- arabinitol dehydrogenase gene on the chromosome of devaromysis Hanseni and cloned into a vector used in E. coli.
  • FIG. 3 is a view showing a method for producing an expression vector comprising the L- arabinitol dehydrogenase gene derived from the devaromysis Hanseni strain.
  • 5 is a diagram showing the appropriate enzyme concentration of L- arabinitol dehydrogenase in the method for producing L-ribulose using the coenzyme solution obtained from the devaromysis Hanseni strain.
  • FIG. 6 is a diagram showing the results of synthesizing L-ribulose using a crude enzyme solution obtained from the devaromysis Hanseni strain.
  • Figure 7 is a diagram showing the production of L-ribulose using the crude enzyme solution obtained from the devaromysis Hanseni strain.
  • the ITS-5.8S rDNA sequence was analyzed at the Korea Microbial Conservation Center. As a result of analyzing the soft relationship with the similar species of the ITS-5.8S rDNA sequence of the S8 strain, it was identified as devaromysis hanseni.
  • the S8 strain was named Debaromyces hansenii KMJ1016, and was deposited internationally in accordance with the Budapest Treaty with Access No. KCCM-10987P on January 30, 2009, to the Korea Microorganism Conservation Center.
  • the yeast bacterium devaromysis hanseni was used to obtain the nucleotide sequence of the L-arabinitol dehydrogenase gene.
  • genes with similar functions are known to be somewhat similar in size to each nucleotide sequence. Therefore, it is estimated that the gene of L-arabinitol dehydrogenase of Devaromysis Hanseni had a size of about 1.0 kb and based on the known L-arabinitol dehydrogenase sequence of other yeasts.
  • the entire gene of L-arabinitol dehydrogenase of Mysis Hanseni was cloned.
  • E. coli XL1-Blue and pUC18 vectors were used for cloning.
  • a culture medium of E. coli LB medium having a general composition was used, and the peptone agar medium (Malt extract peptone agar) was used for culturing devaromysis hanseni.
  • a plate medium of E. coli agar plates containing LB agar, 3-5% sugar, 0.3-0.5% beef extract, 0.9-1.1% bactopeptone, and 1.3-1.7% agar composition were used. 50 ⁇ g / ml ampicillin was added as needed.
  • the culture method was inoculated in a 250 ml Erlenmeyer flask containing 50 ml of devaromysis hanseni and incubated at 37 ° C. and 200 rpm for 1 day, and for E. coli 16 at 37 ° C. and 200 rpm. Time incubation.
  • RNA extraction of devaromysis hanseni was performed using Qiagen plant total RNA kit (QIAGEN), and the reverse transcriptase for cDNA synthesis was Oligo-dT RT-mix (intron).
  • the devaromysis hanseni chromosome was isolated to clone the L-aravinitol dehydrogenase gene.
  • DhLAD F-5 a nonspecific primer based on the L-arabinitol dehydrogenase sequence already known in other yeasts to amplify a portion of the devaromysis hanseni L-arabinitol dehydrogenase gene '-5'- TCA AWB GTR CAG GTM KST GTV GTT CRG MYA TTC AC-3' (SEQ ID NO: 1) and DhLAD R-5'- GAT CAH RMA AGG GAM TTT BYT GGA VCT CKM TAC CAA -3 '(SEQ ID NO: 2) was produced. Using this, a portion of the L-arabinitol dehydrogenase gene corresponding to 730 bp was amplified in the devaromysis hanseni chromosome by a chain poly
  • the genomic DNA of devaromysis hanseni was completely cleaved using restriction enzymes BamHI, EcoRI, HindIII, SalI, and XbaI which do not have a cleavage site in the amplified partial sequences. And a radiolabeled probe was made using a DNA fragment obtained through the polymerase chain reaction. Using this, the DNA fragment containing the gene to be searched by Southern hybridization was searched (FIG. 1). When the chromosome was cut with BamHI, HindIII, and XbaI, the size of the DNA containing the L-arabinitol dehydrogenase gene as a result of Southern hybridization was about 20-23 kb and was not used because it was too large.
  • the desired gene was searched using a fragment cut with EcoRI of about 2.5 kb and SalI of about 5.5 kb.
  • the penicillium pinopylum chromosome was digested with EcoRI, separated from the 2.5 kb DNA fragment and the 5.5 kb DNA fragment isolated from SalI, cloned into pUC18 and named pUC-LAD (FIG. 2).
  • Colony hybridization was performed using the 1.0 kb probe made in the pUC-LAD library to determine clones with the genes of the desired L-arabinitol dehydrogenase.
  • the nucleotide sequence was analyzed using the determined clone, and the total gene nucleotide sequence of L-arabinitol dehydrogenase was found to be 1,056 bp (SEQ ID NO: 3). As expected, it was similar in size to the L-arabinitol dehydrogenase gene found in several other yeasts.
  • Debaromysis Hanseni L-Arabinitol dehydrogenase was found to have the nucleotide sequence common to other L-arabinitol dehydrogenase.
  • the expression vector pGEX-KG (ATCC, USA) BamHI was used to express large amounts of L-arabinitol dehydrogenase in Escherichia coli using a gene encoding L-arabinitol dehydrogenase according to Example 3.
  • the enzyme gene was inserted into the HindIII site and transformed into Escherichia coli BL21 (DE3) (NEB, UK) (FIG. 3).
  • the recombinant strain prepared in Example 4 was inoculated in LB medium and incubated at 37 ° C. for 24 hours, and then the protein expressed in the SDS-PAGE gel was confirmed (FIG. 4).
  • the recombinant strain culture solution was centrifuged (8000 ⁇ g, 10 minutes) to collect only the cells, and then subjected to sonication to the cell wall of E. coli.
  • the supernatant was heat-treated at 70 ° C. for 15 minutes, centrifuged at 20,000 ⁇ g for 20 minutes to remove precipitates, and then the supernatant was obtained.
  • column chromatography was performed using GSTrap (GE Healthcare, Sweden). The recombinant L-arabinitol dehydrogenase was purified purely.
  • Example 5 The physicochemical properties of L-arabinitol dehydrogenase isolated in Example 5 were investigated, and L-arabitol was used as a substrate.
  • the enzyme reaction experiment was carried out under the following conditions.
  • LAD-Arabitol was used as a substrate, and NAD + and NADP + were added as coenzymes.
  • the bacterial culture and enzyme purification were performed in the same manner as in Example 3, and the enzyme and the substrate were reacted at a temperature of 25 ° C. in the presence of 25 mM L-arabitol substrate solution, 1 mM NAD + , and 1 mM NADP + .
  • Example 7 L-ribulose production method using novel L-aravinitol dehydrogenase
  • Example 7-1 Optimum Temperature for L-Riboulose Production
  • Example 5 Production of L-ribulose using L-arabinitol dehydrogenase purely isolated in Example 5 was carried out under the following conditions.
  • the ratio of L-arabitol and L-ribulose was confirmed while changing the temperature.
  • LB As microbial production medium, LB was used.
  • enzyme production medium glycerol 10g L -1 , peptone 1g L -1 , yeast extract 30g L -1 , potassium diphosphate 0.14g L -1 , sodium monophosphate 1g L -1 Added medium was used.
  • absorbance at 600 nm was 0.6
  • 0.1 mM ITPG was added to induce enzyme production.
  • Stirring speed during the process was 200 rpm, the culture temperature was maintained at 37 °C.
  • Example 7-2 Optimal Enzyme Concentration for L-Riboulose Production
  • the L-arabinitol dehydrogenase titration enzyme concentration in L-ribulose production was 15 units ml ⁇ 1 .
  • the production amount of L-ribulose was confirmed while changing the concentration of the initial substrate.
  • Bacterial culture and enzyme purification were performed in the same manner as in Example 3, and the amount of L-ribulose produced was determined using the concentration of L-arabinitol as a substrate in the presence of 1 mM Mg 2+ and 1 mM NAD + as 10-100 g L -1 . Measured. The results are shown in FIG.
  • the productivity of L-ribulose was 57 g L ⁇ 1 h ⁇ 1
  • the conversion of L-ribulose from L-arabitol was 63.1%
  • the production concentration was 63 g L ⁇ 1 .
  • L-ribulose produced by the L-arabinitol dehydrogenase of the present invention is an intermediate of L-ribose synthesis, and L-ribose is L-ribonucleoside, L-oligoribonucleoside and many other treatments. It is an important core pentose that constitutes the skeleton in the synthesis of solvents. The nucleosides derived here have shown considerable potential as useful antiviral agents. Therefore, the conversion of L-arabitol to L-ribulose will play an important role as a basic step for the production of L-ribose.

Landscapes

  • Organic Chemistry (AREA)
  • Chemical & Material Sciences (AREA)
  • Engineering & Computer Science (AREA)
  • Zoology (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Wood Science & Technology (AREA)
  • Chemical Kinetics & Catalysis (AREA)
  • Microbiology (AREA)
  • General Chemical & Material Sciences (AREA)
  • Biotechnology (AREA)
  • Health & Medical Sciences (AREA)
  • Biochemistry (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Enzymes And Modification Thereof (AREA)

Abstract

La présente invention concerne, d'une part une L-arabinitol déshydrogénase exprimée par le gène de la nouvelle L-arabinitol déshydrogénase, d'autre part un procédé de production de L-arabinitol déshydrogénase à partir de souches transformées avec un vecteur d'expression recombinant contenant ledit gène, et aussi un procédé de production de L-ribulose au moyen de ladite déshydrogénase. La L-arabinitol déshydrogénase recombinante de la présente invention, qui transforme spécifiquement le seul L-arabinitol, constitue un catalyseur stable pour une réaction enzymatique. Le procédé de l'invention permet ainsi une production massive et efficace de L-ribulose à partir d'une L-arabinitol déshydrogénase, dérivée en l'occurrence de Debaryomyces hansenii.
PCT/KR2009/000544 2009-02-04 2009-02-04 Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase WO2010090359A1 (fr)

Priority Applications (1)

Application Number Priority Date Filing Date Title
PCT/KR2009/000544 WO2010090359A1 (fr) 2009-02-04 2009-02-04 Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
PCT/KR2009/000544 WO2010090359A1 (fr) 2009-02-04 2009-02-04 Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase

Publications (1)

Publication Number Publication Date
WO2010090359A1 true WO2010090359A1 (fr) 2010-08-12

Family

ID=42542240

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/KR2009/000544 WO2010090359A1 (fr) 2009-02-04 2009-02-04 Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase

Country Status (1)

Country Link
WO (1) WO2010090359A1 (fr)

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108866120A (zh) * 2017-05-10 2018-11-23 韩国科学技术院 基于l-阿拉伯糖的l-核糖的制作方法

Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5766874A (en) * 1991-07-12 1998-06-16 Syntex (Usa) Inc. Kit containing d-arabinitol dehydrogenase and NAD+ for determining d-arabinitol
WO2002066616A2 (fr) * 2001-02-16 2002-08-29 Valtion Teknillinen Tutkimuskeskus Genie de champignons destines a l'utilisation de l-arabinose
JP2005176602A (ja) * 2001-12-27 2005-07-07 National Institute Of Advanced Industrial & Technology 麹菌遺伝子
US20060110809A1 (en) * 2004-05-19 2006-05-25 Biotechnology Research And Development Corp. And Agricultural Research Serv., U.S. Dept. Of Agric. Methods for production of xylitol in microorganisms
US20070259407A1 (en) * 2003-09-12 2007-11-08 Ritva Verho Enzyme for an in Vivo and in Vitro Utilisation of Carbohydrates

Patent Citations (5)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
US5766874A (en) * 1991-07-12 1998-06-16 Syntex (Usa) Inc. Kit containing d-arabinitol dehydrogenase and NAD+ for determining d-arabinitol
WO2002066616A2 (fr) * 2001-02-16 2002-08-29 Valtion Teknillinen Tutkimuskeskus Genie de champignons destines a l'utilisation de l-arabinose
JP2005176602A (ja) * 2001-12-27 2005-07-07 National Institute Of Advanced Industrial & Technology 麹菌遺伝子
US20070259407A1 (en) * 2003-09-12 2007-11-08 Ritva Verho Enzyme for an in Vivo and in Vitro Utilisation of Carbohydrates
US20060110809A1 (en) * 2004-05-19 2006-05-25 Biotechnology Research And Development Corp. And Agricultural Research Serv., U.S. Dept. Of Agric. Methods for production of xylitol in microorganisms

Non-Patent Citations (2)

* Cited by examiner, † Cited by third party
Title
DATABASE GENBANK 10 September 2008 (2008-09-10), "DEHA2C01034p [Debaryomyces hansenii]", retrieved from http://www.ncbi.nlm.nih.gov/protein/199430431 Database accession no. CAG85752 *
DATABASE GENBANK 16 May 2006 (2006-05-16), "hypothetical protein DEHAOCOl l l lg [Debaryomyces hansenii CBS767]", XP000457724, retrieved from http://www.ncbi.nlm.nih.gov/protein/50417778 Database accession no. XP 457724 *

Cited By (1)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
CN108866120A (zh) * 2017-05-10 2018-11-23 韩国科学技术院 基于l-阿拉伯糖的l-核糖的制作方法

Similar Documents

Publication Publication Date Title
WO2018004310A1 (fr) Nouvelle phosphatase résistant à la chaleur, spécifique de l'acide tagatose 6-phosphorique et procédé de préparation de tagatose l'utilisant
JPS63202389A (ja) ２−ケト−ｌ−グロン酸の微生物学的製造方法
EP2356246B1 (fr) Préparation biocatalytique de 5-méthyle uridine
KR100882418B1 (ko) 이노신을 생산하는 미생물 및 그를 이용한 이노신 제조방법
CN115927513A (zh) 一种生物酶制备β-烟酰胺单核苷酸的方法
CN111269870A (zh) 一种高产胞苷酸的重组大肠杆菌及其应用
WO2019112368A1 (fr) Nouvelle psicose-6-phosphate phosphatase, composition de production de psicose comprenant celle-ci, et procédé de production de psicose faisant appel à celle-ci
WO2019098723A1 (fr) Nouvelle d-psicose-3-épimérase et procédé de production de d-psicose à l'aide de celle-ci
WO2011149248A2 (fr) Procédé de préparation d'acide 3-hydroxypropionique à partir de glycérol avec un rendement élevé
CN116515869B (zh) 一种2,4-二氨基丁酸转移酶突变体基因及其应用
WO2010090359A1 (fr) Nouvelle l-arabinitol déshydrogénase, et procédé de production de l-ribulose au moyen de cette déshydrogénase
CN109295026B (zh) 一种n-脱氧核糖转移酶ii定向进化改造和生物催化应用
CN116656585A (zh) 一种促进胞苷合成的方法及应用
WO2014171635A1 (fr) D-sorbitol déshydrogénase inédite et procédé de production de l-sorbose au moyen de celle-ci
CN106834176B (zh) 一种核苷磷酸化酶、编码基因及其高产菌株和应用
WO2017164616A1 (fr) Procédés de préparation de 3'-amino -2',3'-didésoxyguanosine à l'aide de nucléoside phosphorylases dérivées de bacillus et d'adénosine désaminase dérivées de lactococcus
WO2014171636A1 (fr) Nouvelle ribitol déshydrogénase et procédé pour préparer de la l-ribulose l'utilisant
WO2015137565A1 (fr) Nouvelle formaldéhyde déshydrogénase et procédé de préparation de formaldéhyde correspondant
WO2011099657A1 (fr) Xylitol déshydrogénase inédite et procédé de production de l-xylulose au moyen de celle-ci
WO2017164615A1 (fr) Procédé de préparation de 3'-amino-2',3'-didésoxyadénosine au moyen d'une nucléoside phosphorylase dérivée de bacillus
WO2014171570A2 (fr) Production de l-xylulose par couplage d'une nouvelle nadh oxydase dérivée de streptococcus pyogenes et d'une l-arabinitol oxydase
CN115537406B (zh) 一种酮还原酶及其在制备(s)-1-(4-吡啶基)-1,3-丙二醇中的应用
CN115537405B (zh) 一种酮还原酶及其在制备(s)-1-(3-氯苯基)-1,3-丙二醇中的应用
CN114875087B (zh) 以β-吲哚基丙氨酸为底物合成5-羟基β-吲哚基丙氨酸的方法及其应用
EP0263716A2 (fr) Procédé de préparation de l'acide 5'-guanylique et vecteurs d'expression appropriés

Legal Events

Date Code Title Description
121 Ep: the epo has been informed by wipo that ep was designated in this application

Ref document number: 09839727

Country of ref document: EP

Kind code of ref document: A1

NENP Non-entry into the national phase

Ref country code: DE

122 Ep: pct application non-entry in european phase

Ref document number: 09839727

Country of ref document: EP

Kind code of ref document: A1