Engineering of multiple modules to enhance lignocellulose degradation ability in <i>Bacillus subtilis</i> using CRISPR/Cas9 system.

ABSTRACT: Bacillus subtilis is widely employed for lignocellulose degradation. However, wild-type strains typically exhibit low and incomplete cellulase activities. This study aimed to engineer recombinant B. subtilis strains harboring complete, high-efficiency cellulase systems using CRISPR/Cas9-mediated genome editing. After optimization of signal peptides, transcriptional terminators, and chromosomal integration sites, two endoglucanase expression cassettes were integrated into B. subtilis 168, yielding a strain with a 16.29-fold increase in endoglucanase activity relative to the parental strain. In parallel, a bifunctional cellulase was successfully expressed and optimized, achieving exoglucanase (549.77 U/mL) and β-glucosidase (349.26 U/mL) activities. Moreover, the strain

SUBMITTER: Wang Z 

PROVIDER: S-EPMC12907642 | biostudies-literature | 2026 Sep

REPOSITORIES: biostudies-literature