Enhanced trypsin thermostability in Pichia pastoris through truncating the flexible region.

ABSTRACT:

Background

High thermostability is required for trypsin to have wider industrial applications. Target mutagenesis at flexible regions has been proved to be an efficient protein engineering method to enhance the protein thermostability.

Results

The flexible regions in porcine trypsin were predicted using the methods including molecular dynamic simulation, FlexPred, and FoldUnfold. The amino acids 78-90 was predicted to be the highly flexible region simultaneously by the three methods and hence selected to be the mutation target. We constructed five variants (D3, D5, D7, D9, and D11) by truncating the region. And the variant D9 showed higher thermostability, with a 5 °C increase in T_opt, 5.8 °C rise in [Formula: see text], and a 4.5 °C rise in T_m, compare

SUBMITTER: Liu L 

PROVIDER: S-EPMC6201580 | biostudies-literature | 2018 Oct

REPOSITORIES: biostudies-literature