Crystal structures of a meta‐cleavage product hydrolase from Pseudomonas fluorescens IP01 (CumD) complexed with cleavage products

• DOI: 10.1110/ps.0209602
• Publication Date: 2002-10-01T19:26:48Z
• Authors: Shinya Fushinobu; Takashi Saku; Masafumi Hidaka; So‐Young Jun; Hideaki Nojiri; Hisakazu Yamane; Hirofumi Shoun; Toshio Omori; Takayoshi Wakagi
• Keywords: Pseudomonas fluorescens; Hydrolase; Oxyanion hole

2-Hydroxy-6-oxo-7-methylocta-2,4-dienoate hydrolase (CumD) from Pseudomonas fluorescens IP01 hydrolyzes a meta-cleavage product generated in the cumene (isopropylbenzene) degradation pathway. The crystal structures of inactive S103A mutant CumD enzyme complexed with isobutyrate and acetate ions were determined at 1.6 2.0 A resolution, respectively. located same position active site, occupied site for part hydrolysis CumD, which has key determinant group substrate specificity related hydrolases. One oxygen atoms carboxyl ion was hydrogen bonded water molecule His252. Another atom situated an oxyanion hole formed by two main-chain N atoms. isopropyl isobutyric acid recognized side-chains hydrophobic residues. substrate-binding pocket long, inhibition constants various organic acids corresponded well to it. In comparison structure BphD Rhodococcus sp. RHA1, structural basis hydrolases, is revealed.