Highly thermostable carboxylic acid reductases generated by ancestral sequence reconstruction.

ABSTRACT: Carboxylic acid reductases (CARs) are biocatalysts of industrial importance. Their properties, especially their poor stability, render them sub-optimal for use in a bioindustrial pipeline. Here, we employed ancestral sequence reconstruction (ASR) - a burgeoning engineering tool that can identify stabilizing but enzymatically neutral mutations throughout a protein. We used a three-algorithm approach to reconstruct functional ancestors of the Mycobacterial and Nocardial CAR1 orthologues. Ancestral CARs (AncCARs) were confirmed to be CAR enzymes with a preference for aromatic carboxylic acids. Ancestors also showed varied tolerances to solvents, pH and in vivo-like salt concentrations. Compared to well-studied extant CARs, AncCARs had a T_m up to 35 °C higher, with hal

SUBMITTER: Thomas A 

PROVIDER: S-EPMC6874671 | biostudies-literature | 2019

REPOSITORIES: biostudies-literature