Enhancing subtilisin thermostability through a modified normalized B-factor analysis and loop-grafting strategy.

ABSTRACT: Rational design-guided improvement of protein thermostability typically requires identification of residues or regions contributing to instability and introduction of mutations into these residues or regions. One popular method, B-FIT, utilizes B-factors to identify unstable residues or regions and combines them with other strategies, such as directed evolution. Here, we performed structure-based engineering to improve the thermostability of the subtilisin E-S7 (SES7) peptidase. The B-value of each residue was redefined in a normalized B-factor calculation, which was implemented with a refined bioinformatics analysis strategy to identify the critical area (loop 158-162) related to flexibility and to screen for suitable thermostable motif sequences in the Protein Data Bank that can act as t

SUBMITTER: Tang H 

PROVIDER: S-EPMC6885650 | biostudies-literature | 2019 Nov

REPOSITORIES: biostudies-literature