An enantioselective amidase from Burkholderia multivorans for the stereoselective synthesis of esfenvalerate.
Using racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide, an intermediate for the chiral pyrethroid insecticide Esfenvalerate, as a sole nitrogen source in a minimal medium, several strains with high enatioselectivity (≥98%) were isolated by enrichment techniques. One of the strains, LG 31-3, was identified as Burkholderia multivorans, based on physiological and morphological tests by a standardized Biolog station for carbon source utilization. A novel amidase was purified from B. mutivorans LG 31-3 and characterized. The enzyme exhibited (S)- selective amidase activity on racemic (R,S)-2-(4-chlorophenyl)-3-methylbutyramide. Addition of the racemic amide induced the production of the enantioselective amidase. The molecular mass of the amidase on SDS-PAGE analysis was shown to be 50 kDa. The purified amidase was subjected to proteolytic digestion with a modified trypsin. The N-terminal and internal amino acid sequences of the purified amidase showed a high sequence homology with those deduced from a gene named YP_366732.1 encoding indole acetimide hydrolase from Burkholderia sp. 383.
Department of Biotechnology, Korea University Graduate School, Seoul 136-713, Republic of Korea, R&D Center, GS Caltex Corporation, Daejeon 305-380, Republic of Korea.
Sang-Hyun
SH
Lee
Oh-Jin
OJ
Park
Hyun-Jae
HJ
Shin
D000581:Amidohydrolases / Q000737:chemistry* / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000737:chemistry / Q000235:genetics / Q000302:isolation & purification / Q000378:metabolism*; D019117:Burkholderia / Q000201:enzymology* / Q000235:genetics; D008969:Molecular Sequence Data; D009570:Nitriles / Q000737:chemistry / Q000378:metabolism*; D011722:Pyrethrins / Q000737:chemistry / Q000378:metabolism*; D013237:Stereoisomerism
D001426:Bacterial Proteins; D009570:Nitriles; D011722:Pyrethrins; D000581:Amidohydrolases; C019665:amidase; C017690:fenvalerate