Cloning, sequencing, and expression of the gene encoding 4-hydroxy-4-methyl-2-oxoglutarate aldolase from Pseudomonas ochraceae NGJ1

Article abstract of DOI:10.1271/bbb.65.2701

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.

Full text of DOI:10.1271/bbb.65.2701

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Cloning, Sequencing, and Expression of the Gene
Encoding 4-Hydroxy-4-methyl-2-oxoglutarate
Aldolase from Pseudomonas ochraceae NGJ1
Kiyofumi MARUYAMAa, Michiko MIWAa, Nobuyuki TSUJIIa, Tomoyuki NAGAIa, Naotaka
TOMITAa, Toshiyuki HARADAa, Hirobumi SOBAJIMAa & Hiroyuki SUGISAKIb
a Department of Biomolecular Science, Faculty of Engineering, Gifu University
b Institute for Chemical Research, Kyoto University
Published online: 22 May 2014.
To cite this article: Kiyofumi MARUYAMA, Michiko MIWA, Nobuyuki TSUJII, Tomoyuki NAGAI, Naotaka TOMITA, Toshiyuki
HARADA, Hirobumi SOBAJIMA & Hiroyuki SUGISAKI (2001) Cloning, Sequencing, and Expression of the Gene Encoding
4-Hydroxy-4-methyl-2-oxoglutarate Aldolase from Pseudomonas ochraceae NGJ1, Bioscience, Biotechnology, and
Biochemistry, 65:12, 2701-2709, DOI: 10.1271/bbb.65.2701
To link to this article: http://dx.doi.org/10.1271/bbb.65.2701
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