An  in vitro  screening assay to discover novel inhibitors of 4‐coumarate:CoA ligase | Zendy

Yun MinSoo | Zendy; Chen Weijun | Zendy; Deng Fan | Zendy; Kiyokawa Takahiro | Zendy; Mametsuka Koki | Zendy; Yogo Yasuhiro | Zendy

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An in vitro screening assay to discover novel inhibitors of 4‐coumarate:CoA ligase

Author(s) -

Yun MinSoo,

Chen Weijun,

Deng Fan,

Kiyokawa Takahiro,

Mametsuka Koki,

Yogo Yasuhiro

Publication year - 2006

Publication title -

pest management science

Language(s) - English

Resource type - Journals

SCImago Journal Rank - 1.296

H-Index - 125

eISSN - 1526-4998

pISSN - 1526-498X

DOI - 10.1002/ps.1277

Subject(s) - dna ligase , biochemistry , enzyme , high throughput screening , phenylpropanoid , biology , escherichia coli , in vitro , recombinant dna , biosynthesis , gene

4‐Coumarate:CoA ligase (4CL, EC 6.2.1.12) exists only in plants and plays an important role in the phenylpropanoid pathway. Identification of inhibitors targeting 4CL provides a novel approach for developing effective plant growth inhibitors (PGIs). The full‐length gene of tobacco 4CL (Nt4CL1) was cloned and expressed in Escherichia coli Cast & Chalm. The recombinant 4CL protein was extracted and purified by several purification steps including gel‐filtration and anion‐exchange chromatography. 4CL activity assay was miniaturized and optimized using a 96‐well microplate and a reader. Among 28 existing herbicides, propanil and swep strongly inhibited in vitro 4CL enzyme activity, and they were selected for further studies. The process of this assay can be developed into a high‐throughput screening system of PGI targeting 4CL in the phenylpropanoid pathway. Copyright © 2006 Society of Chemical Industry

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