Characterisation of a catabolic epoxide hydrolase from a Corynebacterium sp.
The epoxide hydrolase (EH) from Corynebacterium sp. C12, which grows on cyclohexene oxide as sole carbon source, has been purified to homogeneity in two steps, involving anion exchange followed by hydrophobic-interaction chromatography. The purified enzyme is multimeric (probably tetrameric) with a subunit size of 32,140 Da. The gene encoding Corynebacterium EH was located on a 3.5-kb BamHI fragment of C12 chromosomal DNA using a DNA probe generated by PCR using degenerate primers based on the N-terminal and an internal amino acid sequence. Sequencing and database comparison of the predicted amino acid sequence of Corynebacterium EH shows that it is similar to mammalian and plant soluble EH, and the recently published sequence of epichlorohydrin EH from Agrobacterium radiobacter AD1 [Rink, R., Fennema, M., Smids, M., Dehmel, U. & Janssen, D. B. (1997) J. Biol. Chem. 272, 14650- 14657), particularly around the catalytic site. All of these proteins belong to the alpha/beta-hydrolase-fold family of enzymes. Similarity to the mammalian microsomal EH is weaker.
Misawa
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E
Department of Biochemistry, Imperial College, London, United Kingdom.
Chan Kwo Chion
C K
CK
Archer
I V
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Woodland
M P
MP
Zhou
N Y
NY
Carter
S F
SF
Widdowson
D A
DA
Leak
D J
DJ
D000595:Amino Acid Sequence; D000818:Animals; D001482:Base Composition; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003062:Codon / Q000235:genetics; D003352:Corynebacterium / Q000201:enzymology* / Q000235:genetics / Q000254:growth & development; D003510:Cyclohexanes / Q000378:metabolism; D053138:Cyclohexenes; D017931:DNA Primers / Q000235:genetics; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D004851:Epoxide Hydrolases / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification*; D005798:Genes, Bacterial; D006801:Humans; D008969:Molecular Sequence Data; D016133:Polymerase Chain Reaction; D017386:Sequence Homology, Amino Acid
D003062:Codon; D003510:Cyclohexanes; D053138:Cyclohexenes; D017931:DNA Primers; D004269:DNA, Bacterial; C007193:cyclohexene oxide; D004851:Epoxide Hydrolases