Affiliation:
1. CSIRO Entomology, Canberra, ACT 2601, Australia
Abstract
ABSTRACT
We isolated a bacterial strain,
Agrobacterium radiobacter
P230, which can hydrolyze a wide range of organophosphate (OP) insecticides. A gene encoding a protein involved in OP hydrolysis was cloned from
A. radiobacter
P230 and sequenced. This gene (called
opdA
) had sequence similarity to
opd
, a gene previously shown to encode an OP-hydrolyzing enzyme in
Flavobacterium
sp. strain ATCC 27551 and
Brevundimonas diminuta
MG. Insertional mutation of the
opdA
gene produced a strain lacking the ability to hydrolyze OPs, suggesting that this is the only gene encoding an OP-hydrolyzing enzyme in
A. radiobacter
P230. The OPH and OpdA proteins, encoded by
opd
and
opdA
, respectively, were overexpressed and purified as maltose-binding proteins, and the maltose-binding protein moiety was cleaved and removed. Neither protein was able to hydrolyze the aliphatic OP malathion. The kinetics of the two proteins for diethyl OPs were comparable. For dimethyl OPs, OpdA had a higher
k
cat
than OPH. It was also capable of hydrolyzing the dimethyl OPs phosmet and fenthion, which were not hydrolyzed at detectable levels by OPH.
Publisher
American Society for Microbiology
Subject
Ecology,Applied Microbiology and Biotechnology,Food Science,Biotechnology
Cited by
286 articles.
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