Improving the thermostability of alpha-amylase by combinatorial coevolving-site saturation mutagenesis.

ABSTRACT:

Background

The generation of focused mutant libraries at hotspot residues is an important strategy in directed protein evolution. Existing methods, such as combinatorial active site testing and residual coupling analysis, depend primarily on the evolutionary conserved information to find the hotspot residues. Hardly any attention has been paid to another important functional and structural determinants, the functionally correlated variation information--coevolution.

Results

In this paper, we suggest a new method, named combinatorial coevolving-site saturation mutagenesis (CCSM), in which the functionally correlated variation sites of proteins are chosen as the hotspot sites to construct focused mutant libraries. The CCSM approach was used to improve the thermal stability of

SUBMITTER: Wang C 

PROVIDER: S-EPMC3478181 | biostudies-literature | 2012 Oct

REPOSITORIES: biostudies-literature