Characterization and affinity purification of juvenile hormone esterase from Bombyx mori.
Juvenile hormone esterase (JHE) from hemolymph of the silkworm moth Bombyx mori was characterized for substrate specificity and inhibitor sensitivity. B. mori JHE hydrolyzed the juvenile hormone surrogate substrate methyl n-heptylthioacetothioate (HEPTAT) more efficiently than p-nitrophenyl acetate and 1-naphthyl acetate substrates widely used to assay total carboxylesterase activity. B. mori JHE was sensitive to 3-octylthio-1,1,1-trifluoro-2-propanone (OTFP), which was developed as a selective inhibitor for lepidopteran JHE, and relatively insensitive to diisopropyl fluorophosphate (DFP), an inhibitor of serine esterases but not of all JHEs. Affinity purification with a trifluoromethyl ketone ligand was more efficient for purification of B. mori JHE than DEAE ion exchange chromatography.
Shiotsuki
T
T
Department of Insect Physiology and Behavior, National Institute of Sericultural and Entomological Science, Tsukuba, Ibaraki, Japan. shiotsuk@nises.affrc.go.jp
Bonning
B C
BC
Hirai
M
M
Kikuchi
K
K
Hammock
B D
BD
D000085:Acetates / Q000378:metabolism; D000096:Acetone / Q000031:analogs & derivatives / Q000378:metabolism; D000818:Animals; D012831:Bombyx / Q000201:enzymology*; D002265:Carboxylic Ester Hydrolases / Q000037:antagonists & inhibitors / Q000302:isolation & purification*; D002846:Chromatography, Affinity; D002852:Chromatography, Ion Exchange; D006458:Hemolymph / Q000737:chemistry; D008024:Ligands; D013379:Substrate Specificity
D000085:Acetates; D008024:Ligands; C093097:methyl 1-heptylthioacetothioate; D000096:Acetone; C055697:3-octylthio-1,1,1-trifluoro-2-propanone; D002265:Carboxylic Ester Hydrolases; C038003:juvenile hormone esterase