Characterization and structural analysis of a leucine aminopeptidase using site-directed mutagenesis.

ABSTRACT: Amp0279 (EC 3.4.11.24, GenBank: CP000817.1) is a Co²⁺-dependent leucine aminopeptidase from the Lysinibacillus sphaericus C3-41 genome. After analyses using molecular docking and spatial structure analysis, site-directed mutagenesis mutants were performed as Amp0279-R131E, Amp0279-R131H, Amp0279-R131A and Amp0279-E349D. The optimum pH of Amp0279-R131E was shifted from the original 8.5 to 7.5, and the overall electrostatic potential was shifted towards acidic. Compared with the original enzyme, the mutant proteins all gained better structural stability, especially the apparent melting temperature (T_m) of Amp0279-R131H increased from 41.8 to 45.5 °C. Morever, when protein was bound to the substrate, the T_m of Amp0279-R131E was increased by 7.3 °C and Amp0279-

SUBMITTER: Men Y 

PROVIDER: S-EPMC11655909 | biostudies-literature | 2024 Dec

REPOSITORIES: biostudies-literature