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Mechanism of action and cloning of epoxide hydrolase from the cabbage looper, Trichoplusia ni
Author(s) -
Roe R. Michael,
Kallapur Vasant,
Linderman Russell J.,
Viviani Fabrice,
Harris Shan V.,
Walker Elizabeth A.,
Thompson Deborah M.
Publication year - 1996
Publication title -
archives of insect biochemistry and physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.576
H-Index - 66
eISSN - 1520-6327
pISSN - 0739-4462
DOI - 10.1002/(sici)1520-6327(1996)32:3/4<527::aid-arch24>3.0.co;2-d
Subject(s) - trichoplusia , epoxide hydrolase , microsomal epoxide hydrolase , hydrolase , biology , epoxide , biochemistry , stereochemistry , enzyme , epoxide hydrolase 2 , chemistry , microsome , botany , larva , noctuidae , catalysis
The majority of the JH III epoxide hydrolase activity in last stadium day 3 (gate 1) wandering Trichoplusia ni was membrane bound with approximately 9% of the activity found in the cytosol. Both the microsomal and cytosolic JH epoxide hydrolases were stable, retaining 30% of their original activity after incubation at 4°C for 15 days. 18 O‐labeled water underwent enzyme catalyzed regioselective addition to the least substituted C10 position of JH III. In multiple turnover reactions with JH epoxide hydrolase in 97.9% 18 O‐labeled water, only 91.3% 18 O incorporation was observed. This is consistent with an S N 2 reaction likely involving a carboxylate in the active site of JH epoxide hydrolase. The DNA amplification cloning of a fragment of a putative T. ni epoxide hydrolase is reported. The deduced amino acid sequence shares 67% similarity to the rat microsomal epoxide hydrolase. © 1996 Wiley‐Liss, Inc.