Cloning, Sequencing, and Expression of the Gene Encoding 4-Hydroxy-4-methyl-2-oxoglutarate Aldolase from Pseudomonas ochraceae NGJ1.
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- MARUYAMA Kiyofumi
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- MIWA Michiko
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- TSUJII Nobuyuki
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- NAGAI Tomoyuki
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- TOMITA Naotaka
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- HARADA Toshiyuki
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- SOBAJIMA Hirobumi
- Department of Biomolecular Science, Faculty of Engineering, Gifu University
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- SUGISAKI Hiroyuki
- Institute for Chemical Research, Kyoto University
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Abstract
A DNA fragment that carried the gene (proA) encoding 4-hydroxy-4-methyl-2-oxoglutarate aldolase was cloned from the chromosomal DNA of Pseudomonas ochraceae NGJ1, and the coding region was assigned to the nucleotide sequence based on the N-terminal amino acid sequence of the enzyme purified from the organism. The proA gene was 684 bp long, corresponding to a protein of 227 amino acid residues with a calculated molecular mass of 24,067 Da. The genes encoding a putative transporter and a 4-oxalomesaconate hydratase were upstream, and a 3'-truncated gene encoding 2-pyrone-4,6-dicarboxylate lactonase was downstream from the proA gene in the same orientation on the DNA fragment. The proA gene product was overproduced in Escherichia coli and briefly purified to homogeneity from the crude extract by a two-step purification. The molecular and catalytic properties of the gene product were similar to those of the P. ochraceae enzyme.
Journal
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- Bioscience, Biotechnology, and Biochemistry
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Bioscience, Biotechnology, and Biochemistry 65 (12), 2701-2709, 2001
Japan Society for Bioscience, Biotechnology, and Agrochemistry
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Details 詳細情報について
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- CRID
- 1390282681449577728
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- NII Article ID
- 110002693345
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- NII Book ID
- AA10824164
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- COI
- 1:CAS:528:DC%2BD38XjtVygtQ%3D%3D
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- ISSN
- 13476947
- 09168451
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- NDL BIB ID
- 6027335
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- PubMed
- 11826967
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- Text Lang
- en
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- Data Source
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- JaLC
- NDL
- Crossref
- PubMed
- CiNii Articles
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- Abstract License Flag
- Disallowed