Advancing vitamin B12 production-insights into enhancing yield in Ensifer adhaerens Casida A through ARTP mutagenesis and multi-omics analysis

Ensifer adhaerens, a microorganism recognized for its capacity to synthesize vitamin B12, has garnered significant attention in recent years. Nonetheless, its practical application has been limited by low production yields. Atmospheric and room-temperature plasma mutagenesis was utilized to improve VB12 production and examine the associated mechanisms. Three high-yielding mutant strains BCA-24, BCB-14, and BCC-27 were isolated through multiple rounds of mutagenesis. A multi-omics approach clarified the mechanisms through which these mutations increase VB12 production, including enhanced transcription and translation, optimized energy supply, and improved product efflux. The identification of novel candidate genes in Ensifer adhaerens, which have not been previously studied, provides valuable resources for future genetic engineering aimed at enhancing VB12 production efficiency. This study offers practical improvements in microbial VB12 production while also delivering essential insights into the genetic and metabolic regulation of this important biosynthetic pathway.