Requirement of Catalytic-Triad and Related Amino Acids for the Acyltransferase Activity of Tanacetum cinerariifolium GDSL Lipase/Esterase TcGLIP Involved in Ester-Bond Formation in Pyrethrin Biosynthesis
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概要
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We have recently discovered that a GDSL lipase/esterase (TcGLIP) in Tanacetum cinerariifolium catalyzed acyltransferase activity to form an ester bond in the natural insecticide, pyrethrin. TcGLIP contained Ser40 in Block I, Gly64 in Block II, Asn168 in Block III and Asp318 and His321 in Block V, suggesting underlying hydrolase activity, although little is known about their role in acyltransferase activity. We expressed TcGLIP here in Esherichia coli as a fusion with maltose-binding protein (MBP), part of the fusion being cleaved with a protease to obtain MBP-free TcGLIP. A kinetic analysis revealed that the MBP moiety scarcely influenced the kinetic parameters. The effects on acyltransferase activity of mutations of Gly64, Asn168, Asp318 and His321 were investigated by using MBP-fused TcGLIP. Mutations of these amino acids markedly reduced the acyltransferase activity, suggesting their critical role in the production of pyrethrins.
- 公益社団法人 日本農芸化学会の論文
著者
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TAKEUCHI Takayuki
Department of Internal Medicine, Kyoto Prefectural University of Medicine
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Hatanaka Akikazu
Department Of Agricultural Chemistry Faculty Of Agriculture University Of Yamaguchi
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Matsuda Kazuhiko
Department Of Agricutural Chemistry Kinki University
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Katsuda Yoshio
Dainihon Jochugiku Co. Ltd
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Kikuta Yukio
Dainihon Jochugiku Co. Ltd
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Nakayama Koji
Dainihon Jochugiku Co. Ltd
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YAMADA Gen
Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University
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NAKAYAMA Koji
Dainihon Jochugiku Co., Ltd.
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MITSUMORI Tomonori
Department of Applied Biological Chemistry, Faculty of Agriculture, Kinki University
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