Choi, KJ Yu, YG Hahn, HG Choi, JD Yoon, MY 2024-01-21T04:34:06Z 2024-01-21T04:34:06Z 2021-09-03 2005-08-29 0014-5793 https://pubs.kist.re.kr/handle/201004/136206 Acetohydroxyacid synthase (AHAS) is a thiamin diphosphate- (ThDP-) and FAD-dependent enzyme that catalyzes the first common step in the biosynthetic pathway of the branched-amino acids such as leucine, isoleucine, and valine. The genes of AHAS from Mycobacterium tuberculosis were cloned, and overexpressed in E. coli and purified to homogeneity. The purified AHAS from M. tuberculosis is effectively inhibited by pyrazosulfuron ethyl (PSE), an inhibitor of plant AHAS enzyme, with the IC50 (inhibitory concentration 500/4) of 0.87 mu M. The kinetic parameters of M. tuberculosis AHAS were determined, and an enzyme activity assay system using 96-well microplate was designed. After screening of a chemical library composed of 5600 compounds using the assay system, a new class of AHAS inhibitor was identified with the IC50 in the range of 1.8-2.6 mu M. One of the identified compounds (KHG20612) further showed growth inhibition activity against various strains of M. tuberculosis. The correlation of the inhibitory activity of the identified compound against AHAS to the cell growth inhibition activity suggested that AHAS might be served as a target protein for the development of novel anti-tuberculosis therapeutics. (c) 2005 Published by Elsevier B.V. on behalf of the Federation of European Biochemical Societies. English WILEY ACETOLACTATE SYNTHASE ESCHERICHIA-COLI PURIFICATION RECONSTITUTION SUBUNITS EXPRESSION IMIDAZOLINONES BIOSYNTHESIS HERBICIDES Characterization of acetohydroxyacid synthase from Mycobacterium tuberculosis and the identification of its new inhibitor from the screening of a chemical library Article 10.1016/j.febslet.2005.07.055 1 FEBS LETTERS, v.579, no.21, pp.4903 - 4910 FEBS LETTERS 579 21 4903 4910 scie scopus 000231625800058 2-s2.0-24044548711 Biochemistry & Molecular Biology Biophysics Cell Biology Biochemistry & Molecular Biology Biophysics Cell Biology Article ACETOLACTATE SYNTHASE ESCHERICHIA-COLI PURIFICATION RECONSTITUTION SUBUNITS EXPRESSION IMIDAZOLINONES BIOSYNTHESIS HERBICIDES acetohydroxyacid synthase antibacterial target inhibitor screening antimycobacterial activity