Dissecting the evolvability landscape of the CalB active site toward aromatic substrates.

ABSTRACT: A key event in the directed evolution of enzymes is the systematic use of mutagenesis and selection, a process that can give rise to mutant libraries containing millions of protein variants. To this day, the functional analysis and identification of active variants among such high numbers of mutational possibilities is not a trivial task. Here, we describe a combinatorial semi-rational approach to partly overcome this challenge and help design smaller and smarter mutant libraries. By adapting a liquid medium transesterification assay in organic solvent conditions with a combination of virtual docking, iterative saturation mutagenesis, and residue interaction network (RIN) analysis, we engineered lipase B from P. antarctica (CalB) to improve enzyme recognition and activity against the bulky

SUBMITTER: Santos YLL 

PROVIDER: S-EPMC6821916 | biostudies-literature | 2019 Oct

REPOSITORIES: biostudies-literature