Construction of an efficient electroporation transformation system promotes the application of Targetron in wild-type...

Paenibacillus species have attracted much attention as a promising cell factory that can yield a variety of bioactive peptides and functional exopolysaccharides. Improving the production capacity of Paenibacillus through genetic engineering is crucial for the development of bioactive natural products. This study systematically evaluated the effects of electroporation transformation parameters on the introduction of DNA into Paenibacillus elgii 219, which was isolated from soil and preserved in our lab. The transformation efficiency was improved to 1.25 × 10⁶ transformants/μg DNA by optimizing factors including the growth and recovery media content, the growth stages of P. elgii 219, electric field strength, electroporation buffer composition, cell wall weakening, and DNA quality. The results represent the highest transformation efficiency reported thus far for Paenibacillus. This work further established a group II intron-based gene editing system suitable for P. elgii 219 for the first time. As expected, plasmid pDX4383 was successfully transferred into P. elgii 219, and a putative glycosyltransferase gene was deleted, thereby eliminating flocculent precipitation during fermentation. Taken together, this study demonstrates that the electroporation-mediated transformation of exogenous DNA combined with group II intron-based gene editing provides an effective approach for the genetic engineering of Paenibacillus bacteria.IMPORTANCEThe establishment of an efficient plasmid DNA transformation protocol for P. elgii 219 has established a robust foundation for further in-depth investigations into its physiological and metabolic processes. Additionally, the successful advancement of group II intron-mediated gene editing technology imbues P. elgii 219 with the potential to serve as a highly efficient cell factory. These accomplishments furnish novel perspectives and references for the exploration of other Paenibacillus species in environmental contexts and the development of their products with economic value.