Structure-based directed evolution improves S. cerevisiae growth on xylose by influencing in vivo enzyme performance.

ABSTRACT:

Background

Efficient bioethanol production from hemicellulose feedstocks by Saccharomyces cerevisiae requires xylose utilization. Whereas S. cerevisiae does not metabolize xylose, engineered strains that express xylose isomerase can metabolize xylose by converting it to xylulose. For this, the type II xylose isomerase from Piromyces (PirXI) is used but the in vivo activity is rather low and very high levels of the enzyme are needed for xylose metabolism. In this study, we explore the use of protein engineering and in vivo selection to improve the performance of PirXI. Recently solved crystal structures were used to focus mutagenesis efforts.

Results

We constructed focused mutant libraries of Piromyces xylose isomerase by substitution of second she

SUBMITTER: Lee M 

PROVIDER: S-EPMC6954610 | biostudies-literature | 2020

REPOSITORIES: biostudies-literature