Mining unique cysteine synthetases and computational study on thoroughly eliminating feedback inhibition through tunnel engineering.

ABSTRACT: L-cysteine is an essential component in pharmaceutical and agricultural industries, and synthetic biology has made strides in developing new metabolic pathways for its production, particularly in archaea with unique O-phosphoserine sulfhydrylases (OPSS) as key enzymes. In this study, we employed database mining to identify a highly catalytic activity OPSS from Acetobacterium sp. (AsOPSS). However, it was observed that the enzymatic activity of AsOPSS suffered significant feedback inhibition from the product L-cysteine, exhibiting an IC₅₀ value of merely 1.2 mM. A semi-rational design combined with tunnel analysis strategy was conducted to engineer AsOPSS. The best variant, AsOPSS^A218R was achieved, totally eliminating product inhibition without sacrificing catalytic e

SUBMITTER: Xu S 

PROVIDER: S-EPMC11400630 | biostudies-literature | 2024 Oct

REPOSITORIES: biostudies-literature