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Identification and Biochemical Characterization of a Novel Hormone-Sensitive Lipase Family Esterase Est19 from the Antarctic Bacterium Pseudomonas sp. E2-15.

ABSTRACT: Esterases represent an important class of enzymes with a wide variety of industrial applications. A novel hormone-sensitive lipase (HSL) family esterase, Est19, from the Antarctic bacterium Pseudomonas sp. E2-15 is identified, cloned, and expressed. The enzyme possesses a GESAG motif containing an active serine (S) located within a highly conserved catalytic triad of Ser¹⁵⁵, Asp²⁵³, and His²⁸² residues. The catalytic efficiency (k_cat/K_m) of Est19 for the pNPC6 substrate is 148.68 s^-1mM^-1 at 40 °C. Replacing Glu¹⁵⁴ juxtaposed to the critical catalytic serine with Asp (E154→D substitution) reduced the activity and catalytic efficiency of the enzyme two-fold, with little change in the substrate affinity. The wild-type enzyme retained near complete activity over a temperature range of 10-60 °C, while ~50% of its activity was retained at 0 °C. A phylogenetic analysis suggested that Est19 and its homologs may represent a new subfamily of HSL. The thermal stability and stereo-specificity suggest that the Est19 esterase may be useful for cold and chiral catalyses.

SUBMITTER: Liu X

PROVIDER: S-EPMC8615396 | biostudies-literature |

REPOSITORIES: biostudies-literature

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