WO2002046427A1 - Nouvelle formate deshydrogenase et son procede de production - Google Patents

Nouvelle formate deshydrogenase et son procede de production Download PDF

Info

Publication number
WO2002046427A1
WO2002046427A1 PCT/JP2001/010569 JP0110569W WO0246427A1 WO 2002046427 A1 WO2002046427 A1 WO 2002046427A1 JP 0110569 W JP0110569 W JP 0110569W WO 0246427 A1 WO0246427 A1 WO 0246427A1
Authority
WO
WIPO (PCT)
Prior art keywords
formate dehydrogenase
genus
dna
polypeptide
seq
Prior art date
Application number
PCT/JP2001/010569
Other languages
English (en)
Japanese (ja)
Inventor
Yasuko Takaoka
Hirokazu Nanba
Original Assignee
Kaneka Corporation
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 Kaneka Corporation filed Critical Kaneka Corporation
Priority to JP2002548144A priority Critical patent/JP4287144B2/ja
Priority to AU2002224136A priority patent/AU2002224136A1/en
Publication of WO2002046427A1 publication Critical patent/WO2002046427A1/fr

Links

Classifications

    • C—CHEMISTRY; METALLURGY
    • C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
    • C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
    • C12N9/00—Enzymes; Proenzymes; Compositions thereof; Processes for preparing, activating, inhibiting, separating or purifying enzymes
    • C12N9/0004—Oxidoreductases (1.)
    • C12N9/0008—Oxidoreductases (1.) acting on the aldehyde or oxo group of donors (1.2)

Definitions

  • Formic acid dehydrogenase (enzyme number [EC 1.2.1.2]) is composed of carbon dioxide and reduced nicotine adenine dinucleotide (NADH) from formic acid and oxidized nicotine adenine dinucleotide (NAD). It can be used for coenzyme regeneration in NADH-dependent enzymatic reactions because it catalyzes the reaction that produces methane.In this case, inexpensive formic acid can be used, and by-products are carbon dioxide and do not accumulate in the system. It is a useful enzyme with a point. Furthermore, enzymes with small Km values for formic acid and NAD are effective industrially at low substrate concentrations, and can be used for specific microquantification of formic acid, and are industrially useful enzymes.
  • the present invention is a polypeptide having the following physicochemical properties:
  • the present invention also relates to any one of the following polypeptides (a) to (c): (a) a polypeptide comprising the amino acid sequence shown in SEQ ID NO: 1 in the sequence listing: (b) SEQ ID NO in the sequence listing Polypeptide having one or more amino acids substituted, inserted, deleted and / or or added in the amino acid sequence shown in 1 and having formate dehydrogenase activity:
  • a polypeptide comprising an amino acid sequence having at least 95% homology with the amino acid sequence shown in SEQ ID NO: 1 of the Sequence Listing and having formate dehydrogenase activity.
  • the present invention is also a DNA encoding the above polypeptide.
  • the present invention is also a recombinant plasmid containing the above-mentioned DNA.
  • the present invention also relates to a transformant obtained by transforming a host microorganism with the above DNA or recombinant plasmid.
  • FIG. 1 shows a restriction map of a recombinant plasmid pFAO01 containing a formate dehydrogenase gene according to one embodiment of the present invention.
  • FIG. 2 is a graph showing the Km value for formate of formate dehydrogenase which is one embodiment of the present invention.
  • FIG. 4 shows the range of the action temperature and the optimum temperature of the formate dehydrogenase of one embodiment of the present invention.
  • FIG. 5 shows the range of the action pH and the optimum pH of the formate dehydrogenase of one embodiment of the present invention.
  • H FIG. 6 is a graph showing the temperature stability of formate dehydrogenase which is one embodiment of the present invention.
  • FIG. 7 is a graph showing the pH stability of formate dehydrogenase which is one embodiment of the present invention.
  • the polypeptide of the present invention is a polypeptide having formate dehydrogenase activity, and is characterized by having the following physicochemical properties.
  • NAD is used as a coenzyme to oxidize formic acid to produce carbon dioxide
  • the formate dehydrogenase activity of the polypeptide is determined by the NADH at 30 ° C or 4 ° C in 0.1 M phosphate buffer (I) H7) containing 50 OmM of sodium formate and 5 mM of NAD. This is done by measuring the increase in absorbance at 340 nm with production.
  • the action temperature and the action pH are determined by measuring the activity while changing the temperature or pH under the above-mentioned activity measurement conditions.
  • the temperature stability was determined by measuring the residual activity after treating the polypeptide at each temperature for 10 minutes, and the pH stability was determined by treating the polypeptide at 6 ° C for 20 hours at each pH. It is determined by measuring the residual activity.
  • the molecular weight is determined by gel filtration chromatography.
  • the polypeptide of the present invention can be obtained from a microorganism having formate dehydrogenase activity. Therefore, the microorganism that is the source of the polypeptide of the present invention is methanol-assimilating A bacterium or a formic acid assimilating bacterium is suitable, and is not particularly limited. Examples thereof include microorganisms belonging to the genus Ancylo obacte 1-, and among them, Ancylobacter aquaticus Preferred, more preferably, Ancylobacteraquaticus KNK 607 strain.
  • the aforementioned Ancylobacteraquaticus KNK 607M strain is a strain isolated and obtained by the present inventors in the present invention, and became independent on October 20, 2012 under the accession number FERM BP-7335. It has been deposited at the National Institute of Advanced Industrial Science and Technology (AIST) at the Patent Organism Depositary (Zip code 305-8566, Tsukuba East 1-chome, Ibaraki Pref., Japan 1-6-1 Chuo No. 6) under the Budapest Treaty. The bacteriological properties of Ansilobacter aquaticus (Ancylobabacteraquaticus) KNK607M are shown below. 1. Form
  • Adipic acid one
  • an aqueous medium containing nutrients such as a methanol and nitrogen source as main carbon sources and inorganic salts is used.
  • nutrients such as a methanol and nitrogen source as main carbon sources and inorganic salts
  • organic micronutrients such as vitamins and amino acids often produces favorable results.
  • a nitrogen source ammonia salt, aqueous ammonia, ammonia gas, urea, yeast extract, peptone, Corn's tip liquor and the like are used.
  • the inorganic salts phosphates, magnesium salts, potassium salts, sodium salts, calcium salts, iron salts, sulfates, chlorine and the like are used.
  • the cultivation can usually be performed in the temperature range of 20 ° C to 40 ° C, but preferably 25 ° C to 37 ° C. Further, pH can be cultured at 5.5 to 9.5, but 7 to 9 is preferable. In addition, any of a batch method and a continuous method may be used.
  • the cells are collected from the culture medium by centrifugation or the like after completion of the culture, and the cells are disrupted by means such as ultrasonic disruption to obtain a crude enzyme solution.
  • the polypeptide of the present invention can be obtained by purifying the crude enzyme solution by a salting out method, a column chromatography method, or the like.
  • the polypeptide of the present invention may be a natural enzyme obtained from a microorganism as described above, or may be a recombinant enzyme produced using a gene recombination technique.
  • Examples of the natural enzyme include polypeptide represented by SEQ ID NO: 1 in the sequence listing.
  • polypeptide of the present invention is characterized in that one or several amino acids are substituted, inserted, deleted and / or added in the amino acid sequence shown in SEQ ID NO: 1. It may be a polypeptide comprising an acid sequence and having formate dehydrogenase activity. Further, the polypeptide of the present invention may be a polypeptide having an amino acid sequence having 95% or more homology with the amino acid sequence shown in SEQ ID NO: 1 and having formate dehydrogenase activity. Les ,.
  • amino acid sequence in which one or several amino acids have been substituted, inserted, deleted and / or added is obtained by substituting or inserting an amino acid by a method well known to those skilled in the art such as a partially specific mutagenesis method. Can be obtained by deletion and / or addition. Specifically, it is described in documents such as Nucl e i c A C i d R e s. 10, 6487 (1 982) and Me t od d s in En zymo 1 og y 100, 448 (1 983).
  • a polypeptide having a formate dehydrogenase activity refers to 10% or more, preferably 40%, of the case where a polypeptide consisting of the amino acid sequence shown in SEQ ID NO: 1 is used in the above-mentioned activity measurement conditions. It refers to a polypeptide having an activity of 60% or more, more preferably 80% or more, more preferably.
  • the DNA of the present invention may be a DNA encoding the above polypeptide. It may be the DNA represented by SEQ ID NO: 2 or SEQ ID NO: 3 in the sequence listing, or one or several bases in the base sequence represented by SEQ ID NO: 2 or SEQ ID NO: 3 may be substituted, inserted, deleted, or deleted.
  • the DNA may be a DNA having an added nucleotide sequence. Alternatively, it may be DNA having a nucleotide sequence showing 90% or more, preferably 95% or more homology with the nucleotide sequence shown by SEQ ID NO: 2 or SEQ ID NO: 3.
  • the “base sequence in which one or several bases have been substituted, inserted, deleted and / or added” refers to a protein nucleic acid enzyme extra gene amplification PCR method TAKKAJ 35 (17), 295 1 -3 1 78 (1 990) or Henry A. Er 1 ic li, edited by Ikuyuki Kato, translation, insertion, deletion and Z or addition by methods well known to those skilled in the art as described in PCR Technology 1 (1 990). It means a nucleotide sequence in which a possible number of bases are substituted, inserted, deleted and / or added.
  • the homology between the amino acid sequence and the base sequence is determined by the software Cement (S oft wa re D evelo pme nt) manufactured by softwares der Ru Jieneteikusu (GENETYX) DNA (formate dehydrogenase gene) of the c the present invention that can be calculated by comparing the sequences using the previously described It can be obtained from a microorganism having such a formate dehydrogenase activity. To obtain the target DNA, for example, the following method can be used.
  • the amino acid sequence at the amino terminus of formate dehydrogenase purified from a microorganism having formate dehydrogenase activity is determined using a gas-phase protein sequencer or the like.
  • a DNA primer designed based on this amino acid sequence and a DNA primer designed based on the sequence of a highly homologous portion in the base sequence of a known formate dehydrogenase gene are synthesized.
  • the chromosomal DNA is isolated from the microorganism that is the source of formate dehydrogenase.
  • the chromosomal DNA is obtained by dissolving the cultured cells with lysozyme or a surfactant, depositing the extracted DNA with ethanol, attaching it to a glass rod, and dissolving it in an appropriate buffer. It can be obtained by purification several times repeatedly (for example, see the Marmur method described in Mo. Bio1, 3, 208 (1961)).
  • a part of the target gene can be obtained by performing PCR using this chromosomal DNA as a type III using the above primers.
  • the chromosomal DNA obtained previously with appropriate restriction enzymes such as BamHI and PstI was digested with agarose gel electrophoresis, followed by PCR.
  • Southern hybridization is performed using the obtained DNA fragment of the formate dehydrogenase gene as a probe, and a DNA fragment that contains the formate dehydrogenase gene and is not cleaved by the restriction enzyme in the formate dehydrogenase gene Is detected on the gel.
  • the DNA obtained by cyclization using T4 DNA ligase or the like is converted into type II, and the N-terminal side of the enzyme of the partial formate dehydrogenase gene obtained previously by PCR.
  • a DNA primer is synthesized in the outward direction of the enzyme gene, and PCR is performed using these primers. A DNA fragment encoding the terminal side and the C-terminal side can be obtained.
  • a DNA fragment containing the full length of the formate dehydrogenase gene can be obtained.
  • the obtained DNA fragment is confirmed to contain the full length of the target formate dehydrogenase gene by molecular weight measurement and partial base sequence analysis.
  • a recombinant plasmid can be obtained by ligating the obtained DNA fragment containing the formate dehydrogenase gene with vector DNA using T4 DNA ligase or the like.
  • the nucleotide sequence of the DNA fragment containing the formate dehydrogenase gene inserted into the vector was analyzed to confirm that there was a base encoding the N-terminal amino acid sequence of formate dehydrogenase.
  • the translation initiation site is determined from this, and the base sequence is homologous to the known formate dehydrogenase gene, and the encoded protein matches the protein molecular weight by electrophoresis, using the open codon until the stop codon as an open reading frame. Make sure that it is the target gene.
  • a transformant By using the thus obtained DNA or a recombinant plasmid obtained by incorporating the DNA into a vector, a transformant can be obtained by transforming a host microorganism.
  • hosts include the genus Escherichia, the genus Pseudomonas, the genus Flavobacterium, the genus Bacillus, and Ser1-atia.
  • a microorganism belonging to the genus can be used, and as a vector, a plasmid, phage or a derivative thereof derived from a microorganism capable of autonomous replication in the above host can be used.
  • Escherichiacoli it is preferable to use Escherichiacoli as a host microorganism and a vector capable of autonomous replication in the microorganism as a vector.
  • Escherichiacoli HB101 was transformed using the recombinant plasmid pFAO01 in which the DNA obtained as described above was integrated into the pUC19 vector, Transformant Escherichia coli HB101 (pFA01) can be obtained.
  • the transformant Escherichiacoli HB101 (pFA001) obtained by the present invention was obtained on October 20, 2012 (20Q0) under the accession number F ERM BP-7334. It has been deposited with the National Institute of Advanced Industrial Science and Technology (AIST) at the Patent Organism Depositary (Postal code 305-8566, Tsukuba East 1-chome, Ibaraki, Japan 1-6-1 Central No. 6) under the Budapest Treaty.
  • AIST National Institute of Advanced Industrial Science and Technology
  • a vector modified to have a strong structural promoter to increase the production of the enzyme can be used.
  • Production of formate dehydrogenase by the transformant obtained in the present invention may be carried out by culturing using a normal medium.
  • the medium used for the culture may be a usual medium containing a carbon source, a nitrogen source and nutrients such as inorganic salts. Addition of organic micronutrients such as vitamins and amino acids to this often results in favorable results.
  • Carbon sources include carbohydrates such as glucose and sucrose; organic acids such as acetic acid; Alcohols and the like are appropriately used.
  • As the nitrogen source ammonia salt, ammonia water, ammonia gas, urea, yeast extract, peptone, corn 'tea liquor and the like are used.
  • inorganic salts phosphates, magnesium salts, potassium salts, sodium salts, calcium salts, iron salts, sulfates, chlorine and the like are used.
  • the cultivation can be performed in a temperature range of 25 ° C to 40 ° C, but a temperature of 25 ° C to 37 ° C is particularly preferred.
  • the pH can be cultured at 4 to 8, but is preferably at 5 to 7.5.
  • any of a batch culture method and a continuous culture method may be used.
  • Example 1 Acquisition of Ansilobacter aquaticus KNK607M strain Ancylobacteraquaticus KNK607M strain which produces formate dehydrogenase at a high level of the present invention is isolated as follows. did. Suspension of soil samples collected from various places is suspended in 0.9% saline solution, and the supernatant is inoculated into 10 ml of a liquid medium with the composition shown in Table 1 using methanol as a single carbon source, and inoculated at 1% to 30 °. C. The cells were cultured aerobically with shaking.
  • the cells obtained by centrifuging 7 ml of the culture solution of the selected bacteria were suspended in 0.7 ml of 0.1 M phosphate buffer (pH 7), sonicated, and centrifuged.
  • the supernatant was obtained as a crude enzyme solution.
  • Measurement of formate dehydrogenase activity of the crude enzyme solution was performed using sodium formate 500 mM, NAD The measurement was carried out by measuring the increase in absorbance at 340 nm at 30 ° C. in a 0.1 M phosphate buffer (pH 7) containing the following.
  • the protein was quantified by the Bradford method using BSA as a standard protein (see Anal. Biochem., Vol. 72, 248, 1976).
  • the formate dehydrogenase-active strain was purified from the culture solution by the monocolony method, and the resulting purified strain was cultured in the same manner as described above to prepare a crude enzyme solution. Thereafter, formate dehydrogenase activity and protein concentration were measured. Then, the specific activity of the crude enzyme solution of each strain was compared, and as a high specific activity strain, Ancylobacteraquaticus KNK 607M strain (FERM BP-7334) was obtained.
  • Ancillobacter aquaticus (Ancylobaccteraquaticus) A colony of the KNK 607M strain was inoculated into 10 ml of a medium having a composition excluding formic acid from the composition shown in Table 2, and aerobically shaken at 30 ° C for 3 days. This culture solution was inoculated at 1% of the medium volume into 100 ml of the main culture medium in Table 2 per flask and cultured aerobically with shaking at 30 ° C for 4 days.
  • This fraction was dissolved in a 1 M phosphate buffer (pH 6.5) containing 1 DTT and EDTA, dialyzed with the same buffer, and subjected to DEA ES epharose (Pharmacia) column chromatography. After washing with, elution was performed with 0.1 M phosphate buffer (pH 6.2) containing 1 mM DTT and EDTA, and the active fraction was collected. Ammonium sulfate was added to the mixture to 25% saturation, then applied to TSKgel Phenyl Toyopearl 650M (manufactured by Tosoichi Co., Ltd.), and subjected to column chromatography.
  • the Km values for formic acid and NAD were examined under the conditions for measuring the specific activity (30 ° C.).
  • the Km value of formic acid was measured by changing the concentration of sodium formate under the conditions of NAD 5 mM.
  • the activity was measured under a condition of 500 mM PAD while changing the NAD concentration. From the results shown in FIG. 2, the Km value for formic acid was 2.42 mM, and the Km value for NAD was 0.057 from FIG.
  • the range of action pH and the optimum pH were examined under the conditions for measuring the specific activity (3 CTC).
  • the relative activity at each pH assuming that the activity at pH 6.3 is 100%, is shown in FIG.
  • This enzyme showed a relative activity of 3% or more in the range of pH 5 to 11, and particularly a relative activity of 80% or more in the range of pH 5.5 to 9.5. From this, the action pH was set to 5 to 11, and the optimum pH was set to 5.5 to 9.5.
  • the temperature stability of the enzyme was examined by measuring the residual activity of the enzyme in 0.1 M phosphate buffer (pH 7) after holding at each temperature for 10 minutes. As a result, as shown in FIG. 6, the residual activity was 80% or more at a temperature of 50 ° C. or less, which was stable.
  • the molecular weight was determined by gel filtration chromatography method to be about 1.0 X 10
  • a DNA primer having the sequence (Primer-6) the DNA between the sequences is amplified by PCR to obtain a DNA fragment (SEQ ID NO: 2) containing the entire length of the formate dehydrogenase gene. I got it. From the analysis of the molecular weight and a part of the nucleotide sequence of the obtained DNA fragment, it was confirmed that the full length of the formate dehydrogenase gene (SEQ ID NO: 3 in the Sequence Listing) was included.
  • the DNA fragment containing the formate dehydrogenase gene obtained in Example 4 was cleaved with restriction enzymes SphI and EcoRI, and vector plasmid pUC19 and T4DNA ligase cut with the same enzymes were used.
  • a plasmid pFAO01 represented by the restriction map shown in FIG. 1 and containing the formate dehydrogenase gene was obtained.
  • the FDH gene in the figure represents the formate dehydrogenase gene of the present invention.
  • nucleotide sequence of the DNA fragment containing the full length of the formate dehydrogenase gene obtained in this manner is deduced from the nucleotide sequence in SEQ ID NO: 2 in the Sequence Listing and the nucleotide sequence of the open reading frame in SEQ ID NO: 3 in the Sequence Listing.
  • SEQ ID NO: 1 Pointing out toungue
  • Transformation was performed by mixing the plasmid pFA001 obtained in Example 5 with competent cells of Escherichiacoli HB101 strain, and plating was performed on an agar medium shown in Table 3. Then, a transformant containing the recombinant DNA containing the formate dehydrogenase gene was obtained as a colony. (Table 3)
  • Phosphorus was added after sterilization.
  • the cells are collected from the resulting culture by centrifugation, suspended in 0.1 M phosphate buffer (PH 7), disrupted by ultrasonication, and centrifuged to remove insoluble matter from the cells.
  • 0.1 ml of the obtained enzyme solution was added to 11 ⁇ 4 formic acid 1 ⁇ & (0.1 M phosphate buffer, pH7) 1.5 ml, 0.1 M AD 0.15 ml, 0.1 M phosphoric acid.
  • the present invention Since the present invention has the above-mentioned constitution, it has a high specific activity, a small Km value for formic acid and NAD, a wide range of temperature stability and pH stability, and a wide range of action pH and temperature.
  • a formate dehydrogenase suitable for industrial use and a method for producing the same can be provided.

Landscapes

  • Life Sciences & Earth Sciences (AREA)
  • Chemical & Material Sciences (AREA)
  • Health & Medical Sciences (AREA)
  • Genetics & Genomics (AREA)
  • Organic Chemistry (AREA)
  • Engineering & Computer Science (AREA)
  • Bioinformatics & Cheminformatics (AREA)
  • Zoology (AREA)
  • Wood Science & Technology (AREA)
  • Molecular Biology (AREA)
  • Microbiology (AREA)
  • Biotechnology (AREA)
  • Biomedical Technology (AREA)
  • Biochemistry (AREA)
  • General Engineering & Computer Science (AREA)
  • General Health & Medical Sciences (AREA)
  • Medicinal Chemistry (AREA)
  • Enzymes And Modification Thereof (AREA)
  • Micro-Organisms Or Cultivation Processes Thereof (AREA)

Abstract

L'invention concerne une formate déshydrogénase présentant une activité spécifique élevée et une petite valeur Km pour l'acide formique et la NAD, stable sur une grande plage de température et une grande plage de pH, présentant une activité sur une grande plage de pH et sur une grande plage de température et pouvant être utilisée à des fins industrielles. Un micro-organisme produisant la formate déshydrogénase est recherché dans le sol, ce micro-organisme produisant l'enzyme susmentionnée, laquelle présente des propriétés permettant son utilisation à des fins industrielles. L'invention concerne également un ADN contenant le gène de cette enzyme, un ADN recombiné avec un vecteur, et son transformant préparé à l'aide d'un plasmide. L'invention concerne en outre un procédé de production de cette formate déshydrogénase au moyen d'une souche appartenant au genre Ancylobacter ou d'un transformant, à l'aide d'un gène de formate déshydrogénase provenant de cette souche.
PCT/JP2001/010569 2000-12-04 2001-12-04 Nouvelle formate deshydrogenase et son procede de production WO2002046427A1 (fr)

Priority Applications (2)

Application Number Priority Date Filing Date Title
JP2002548144A JP4287144B2 (ja) 2000-12-04 2001-12-04 新規ギ酸脱水素酵素及びその製造方法
AU2002224136A AU2002224136A1 (en) 2000-12-04 2001-12-04 Novel formate dehydrogenase and process for producing the same

Applications Claiming Priority (2)

Application Number Priority Date Filing Date Title
JP2000368838 2000-12-04
JP2000-368838 2000-12-04

Publications (1)

Publication Number Publication Date
WO2002046427A1 true WO2002046427A1 (fr) 2002-06-13

Family

ID=18838980

Family Applications (1)

Application Number Title Priority Date Filing Date
PCT/JP2001/010569 WO2002046427A1 (fr) 2000-12-04 2001-12-04 Nouvelle formate deshydrogenase et son procede de production

Country Status (3)

Country Link
JP (1) JP4287144B2 (fr)
AU (1) AU2002224136A1 (fr)
WO (1) WO2002046427A1 (fr)

Cited By (6)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003031626A1 (fr) * 2001-10-09 2003-04-17 Kaneka Corporation Nouvelle formiate deshydrogenase tolerante aux composes halogenes et son procede de production
JP2004303601A (ja) * 2003-03-31 2004-10-28 Sharp Corp エネルギー回収システム
WO2007015511A1 (fr) 2005-08-02 2007-02-08 Kaneka Corporation Oxydase d’acide d-aminé, et procédé de production d’acide l-aminé, d’acide 2-oxo ou imine cyclique
WO2010067578A1 (fr) 2008-12-09 2010-06-17 株式会社カネカ Nouvelle déshydrogénase d'acides aminés, et procédé de production d'acide aminé l, d'acide 2-oxo ou d'acide aminé d
US8481294B2 (en) 2009-08-03 2013-07-09 Toyota Jidosha Kabushiki Kaisha Mutant formate dehydrogenase, gene encoding the same, and method for producing NADH
CN115672365A (zh) * 2022-08-29 2023-02-03 福州大学 一种高效温和合成氨催化剂及其制备方法和应用

Non-Patent Citations (5)

* Cited by examiner, † Cited by third party
Title
LOGINOVA, N.V.: "Autotrophic metabolism of methanol in microcyclus aquaticus", MIKROBIOLOGIYA, vol. 47, no. 1, 1978, pages 168 - 170, XP002909473 *
POPOV, V.O.: "NAD(+)-dependent formate dehydrogenase", BIOCHEM. J., vol. 301, no. PART 3, 1994, pages 625 - 643, XP002909477 *
POPOV, V.O.: "NAD-dependent formate dehydrogenase from methylotrophic bacteria pseudomonas sp. 101.1 amino acid sequence", BIOORG. KHIM., vol. 16, no. 3, 1990, pages 324 - 335, XP002909475 *
TISHKOV, V.I.: "Catarytic properties and stability of a pseudomonas sp. 101 formate dehydrogenase mutants containing Cys-255-Ser and Cys-255-Met replacement", BIOCHEM. BIOPHYS. RES. COMMUN., vol. 192, no. 3, 1993, pages 976 - 981, XP002909476 *
URAKAMI, T.: "Electrophoretic comparison of enzymes in the gram negative methanol-utilizing bacteria", J. GEN. APPL. MICROBIOL., vol. 27, no. 5, 1981, pages 381 - 403, XP002909474 *

Cited By (7)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
WO2003031626A1 (fr) * 2001-10-09 2003-04-17 Kaneka Corporation Nouvelle formiate deshydrogenase tolerante aux composes halogenes et son procede de production
US7432095B2 (en) 2001-10-09 2008-10-07 Kaneka Corporation Formate dehydrogenase tolerant to halogen compounds and process for producing the same
JP2004303601A (ja) * 2003-03-31 2004-10-28 Sharp Corp エネルギー回収システム
WO2007015511A1 (fr) 2005-08-02 2007-02-08 Kaneka Corporation Oxydase d’acide d-aminé, et procédé de production d’acide l-aminé, d’acide 2-oxo ou imine cyclique
WO2010067578A1 (fr) 2008-12-09 2010-06-17 株式会社カネカ Nouvelle déshydrogénase d'acides aminés, et procédé de production d'acide aminé l, d'acide 2-oxo ou d'acide aminé d
US8481294B2 (en) 2009-08-03 2013-07-09 Toyota Jidosha Kabushiki Kaisha Mutant formate dehydrogenase, gene encoding the same, and method for producing NADH
CN115672365A (zh) * 2022-08-29 2023-02-03 福州大学 一种高效温和合成氨催化剂及其制备方法和应用

Also Published As

Publication number Publication date
AU2002224136A1 (en) 2002-06-18
JPWO2002046427A1 (ja) 2004-04-08
JP4287144B2 (ja) 2009-07-01

Similar Documents

Publication Publication Date Title
Ansari et al. Recombinant β-galactosidases–Past, present and future: A mini review
JPWO2002068659A1 (ja) アガラーゼおよびその遺伝子
JP3408737B2 (ja) ニトリルヒドラターゼの活性化に関与するタンパク質及びそれをコードする遺伝子
EP1449923B1 (fr) Nouvelle formiate deshydrogenase tolerante aux composes halogenes et son procede de production
Yamamoto et al. Characterization of theanine-forming enzyme from Methylovorus mays no. 9 in respect to utilization of theanine production
JP4287144B2 (ja) 新規ギ酸脱水素酵素及びその製造方法
Nanba et al. Purification and characterization of formate dehydrogenase from Ancylobacter aquaticus strain KNK607M, and cloning of the gene
JP4146095B2 (ja) 耐熱性グルコキナーゼ遺伝子、それを含有する組換えベクター、その組換えベクターを含有する形質転換体及びその形質転換体を用いた耐熱性グルコキナーゼの製造方法
JP2971218B2 (ja) ウリカーゼ遺伝子およびウリカーゼの製造法
JP4880859B2 (ja) 新規カルボニル還元酵素、その遺伝子、およびその利用法
US20050227325A1 (en) Recombinant polynucleotide
JPS63157986A (ja) 遺伝子組換によるｌ−フエニルアラニンの製造方法
JP4627039B2 (ja) アミダーゼ活性を有するポリペプチド及びその遺伝子
JP4245229B2 (ja) 安定なヘキソキナーゼおよびその製造法
JP5183265B2 (ja) 遺伝子高発現系
JP4415247B2 (ja) 新規なグリセロールキナーゼ、該遺伝子及び該遺伝子を用いたグリセロールキナーゼの製造法
JP4161236B2 (ja) 新規なＬ−α−グリセロホスフェートオキシダーゼおよびその製造方法
JP2005102511A (ja) 新規アルコール脱水素酵素、その遺伝子
JP3829950B2 (ja) 新規なクレアチニンアミドヒドロラーゼ
JP2004089052A (ja) グルコース脱水素酵素の製造方法
JP4066203B2 (ja) 新規なクレアチニンアミドヒドロラーゼ
JP2002065270A (ja) 耐熱性フェニルアラニン脱水素酵素遺伝子、それを含有する組換えベクター、その組換えベクターを含有する形質転換体及びその形質転換体を用いた耐熱性フェニルアラニン脱水素酵素の製造方法
JP2003125784A (ja) 新規チロシン脱炭酸酵素
JP2001252088A (ja) クレアチンアミジノヒドロラーゼをコードする遺伝子
JPH04325086A (ja) セラチア属に属する形質転換微生物及びそれを用いるビオチンの製造方法

Legal Events

Date Code Title Description
ENP Entry into the national phase

Ref country code: JP

Ref document number: 2002 548144

Kind code of ref document: A

Format of ref document f/p: F

AK Designated states

Kind code of ref document: A1

Designated state(s): AE AG AL AM AT AU AZ BA BB BG BR BY BZ CA CH CN CO CR CU CZ DE DK DM DZ EC EE ES FI GB GD GE GH GM HR HU ID IL IN IS JP KE KG KP KR KZ LC LK LR LS LT LU LV MA MD MG MK MN MW MX MZ NO NZ OM PH PL PT RO RU SD SE SG SI SK SL TJ TM TR TT TZ UA UG US UZ VN YU ZA ZM ZW

AL Designated countries for regional patents

Kind code of ref document: A1

Designated state(s): GH GM KE LS MW MZ SD SL SZ TZ UG ZM ZW AM AZ BY KG KZ MD RU TJ TM AT BE CH CY DE DK ES FI FR GB GR IE IT LU MC NL PT SE TR BF BJ CF CG CI CM GA GN GQ GW ML MR NE SN TD TG

121 Ep: the epo has been informed by wipo that ep was designated in this application
REG Reference to national code

Ref country code: DE

Ref legal event code: 8642

122 Ep: pct application non-entry in european phase