Exploring the abundance of oleate hydratases in the genus Rhodococcus —discovery of novel enzymes with complementary substrate scope

摘要: Oleate hydratases (Ohys, EC 4.2.1.53) are a class of enzymes capable selective water addition reactions to broad range unsaturated fatty acids leading the respective chiral alcohols. Much research was dedicated improving applications existing Ohys as well identification undescribed with potentially novel properties. This study focuses on latter by exploring genus Rhodococcus for its plenitude oleate hydratases. Three different clades showed presence whereby each clade represented specific hydratase family (HFam). Phylogenetic and sequence analyses revealed HFam-specific patterns amongst conserved amino acids. from two strains (HFam 2 3) were heterologously expressed in Escherichia coli their substrate scope investigated. Here, both complementary behaviour towards sterically demanding multiple Furthermore, this includes characterisation newly discovered pyridinivorans Ohy. The steady-state kinetics R. Ohy measured using coupled assay based alcohol dehydrogenase NAD+-dependent oxidation 10-hydroxystearic acid.