Construction of an efficient electroporation transformation system promotes the application of Targetron in wild-type <i>Paenibacillus elgii</i> 219

<i>Paenibacillus</i> 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 <i>Paenibacillus</i> 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 <i>Paenibacillus elgii</i> 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 <i>P. elgii</i> 219, electric field strength, electroporation buffer composition, cell wall weakening, and DNA quality. The results represent the highest transformation efficiency reported thus far for <i>Paenibacillus</i>. This work further established a group II intron-based gene editing system suitable for <i>P. elgii</i> 219 for the first time. As expected, plasmid pDX4383 was successfully transferred into <i>P. elgii</i> 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 <i>Paenibacillus</i> bacteria.IMPORTANCEThe establishment of an efficient plasmid DNA transformation protocol for <i>P. elgii</i> 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 <i>P. elgii</i> 219 with the potential to serve as a highly efficient cell factory. These accomplishments furnish novel perspectives and references for the exploration of other <i>Paenibacillus</i> species in environmental contexts and the development of their products with economic value.