Coexistence of endonuclease and exonuclease activities in a novel RecJ from Bacillus cereus

Ma, Liya; Wang, Wen; Hao, Chaozhi; Zheng, Lirong; Zheng, Minggang

doi:10.1007/s10529-021-03107-z

Cited by 2 publications

(1 citation statement)

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“…A novel class of RecJ nucleases, distinct from the classic RecJ nucleases and derived from Bacillus alcalophilus, Bacillus cereus, and Bacillus halodurans, exhibit not only the exonuclease activity of RecJ but also endonuclease activity (Ma et al, 2021;Wang et al, 2020;Zheng et al, 2020;Srivastav et al, 2019;Makarova et al, 2012). Based on the combined exo-and endonuclease activities of the RecJ (BaRecJ) nuclease from B. alkaliphilic, we established a novel global in vivo mutagenesis method, differentiating it from current global in vivo mutagenesis mechanisms (Luan et al, 2013;Badran and Liu, 2015).…”

Section: Introductionmentioning

confidence: 99%

Highly efficient in vivo mutagenesis method based on the endonuclease and exonuclease activity of Bacillus alcalophilus RecJ (BaRecJ)

Shang,

Zhang,

et al. 2024

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With the continual advancement of technologies such as microbial cultivation, DNA sequencing, bioinformatics, and genetic engineering, methods for microbial breeding have become increasingly diverse. We identified a RecJ enzyme (BaRecJ) with both endonuclease and exonuclease activities from B. alcalophilus. Differing from traditional physical-chemical mutagenesis approaches and the methods based on perturbation factor. this research established a novel mutagenesis method utilizing the endonuclease and exonuclease activities of BaRecJ. Mutagenesis of E. coli was conducted using the BaRecJ method, followed by screening for rifampicin-resistant mutants, rpoB sequencing results demonstrated a broader, more uniform spectrum of mutations and a higher frequency of substitution mutations with this mutagenesis approach. Furthermore, this mutagenesis method was applied to S cerevisiae, resulting in mutants with enhanced tolerance to acetic acid and ethanol, exhibiting improved fermentation performance and flocculation abilities Genomic resequencing analysis summarized genes possibly associated with the tolerance of mutants. Therefore, this approach 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 genetic loci associated with specific traits.

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