CLONING OF PSEUDOMONAS-FLUORESCENS CARBOXYLESTERASE GENE AND CHARACTERIZATION OF ITS PRODUCT EXPRESSED IN ESCHERICHIA-COLI

A gene (estC) coding for an esterase (esterase III) of Pseudomonas fluorescens was cloned into Escherichia coli JM83. DNA sequencing showed a single open reading frame with GTG as a translation initiation codon for esterase III. This was confirmed by N-terminal amino acid sequence analysis of the purified esterase III protein from an E. coli clone. The promoter sequence and a potential Shine-Dalgarno sequence were followed by the coding sequence of the estC gene. The amino acid sequence deduced from the nucleotide sequence contains the consensus active site sequence, G-X-S-X-G, of serine esterase. The esterase III expressed in an E. coli clone was purified by ion-exchange chromatography and gel filtration. The native form of the enzyme was a monomer with a molecular weight of 41,000. The results of substrate specificity and the inhibitor studies suggest that this enzyme is a carboxylesterase (EC 3.1.1.1) and a serine residue is present at the active site of the enzyme.
Publisher
JAPAN SOC BIOSCI BIOTECHN AGROCHEM
Issue Date
1994-01
Language
ENG
Keywords

TRANSLATIONAL INITIATION; INSECTICIDE RESISTANCE; INTRACELLULAR ESTERASE; NUCLEOTIDE-SEQUENCE; PURIFICATION; LIPASE; MEMBRANE; PROTEINS

Citation

BIOSCIENCE BIOTECHNOLOGY AND BIOCHEMISTRY, v.58, no.1, pp.111 - 116

ISSN
0916-8451
URI
http://hdl.handle.net/10203/6156
Appears in Collection
MSE-Journal Papers(저널논문)
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