Recombining low homology, functionally rich regions of bacterial subtilisins by combinatorial fragment exchange.

ABSTRACT: Combinatorial fragment exchange was utilised to recombine key structural and functional low homology regions of bacilli subtilisins to generate new active hybrid proteases with altered substrate profiles. Up to six different regions comprising mostly of loop residues from the commercially important subtilisin Savinase were exchanged with the structurally equivalent regions of six other subtilisins. The six additional subtilisins derive from diverse origins and included thermophilic and intracellular subtilisins as well as other academically and commercially relevant subtilisins. Savinase was largely tolerant to fragment exchange; rational replacement of all six regions with 5 of 6 donating subtilisin sequences preserved activity, albeit reduced compared to Savinase. A combinatorial approac

SUBMITTER: Jones DD 

PROVIDER: S-EPMC3168465 | biostudies-literature | 2011

REPOSITORIES: biostudies-literature