Bacillus alcalophilus RecJ (BaRecJ) can drive the adaptive evolution of S. cerevisiae.

With the continuous advancement of technologies such as microbial cultivation, DNA sequencing, bioinformatics, and genetic engineering, in vivo mutagenesis methods based on perturbation factors are now widely utilized. We identified a RecJ enzyme (BaRecJ) with endonuclease and exonuclease activities from Bacillus alcalophilus, and established a broad-spectrum mutagenic method based on the endonuclease and exonuclease activities of BaRecJ. The BaRecJ mutagenesis method was applied to S cerevisiae to enhance its ethanol or acetic acid tolerance, resulting in mutant strains with improved fermentation performance. Genomic resequencing analysis summarized genes possibly associated with the tolerance of mutants. BaRecJ mutagenesis method not only holds immense potential in microbial mutagenesis breeding and adaptive evolution but also, when coupled with genomic resequencing, allows for the rapid identification of candidate genetic loci associated with specific traits.