Characterization of the Cell-Surface Barriers to Plasmid Transformation in Corynebacterium glutamicum

The effects of including glycine and isonicotinic acid hydrazide (INH) in the growth medium (Luria broth, LBG) on the subsequent lysozyme-imduced protoplast formation and transformation efficiency of Corynebacterium glutamicum were studied. The transformation efficiency of C. glutamicum AS019 increased up to 100-fold as the ocncentrationof glycine in the media increased from 0% to 5% (w/v), relative to cells grown in the absence of glycine. The presence of 5 mg/ml INH in the growth medium led to a further 10-fold increase in transformation efficiency. In addition, this transformation protocol was successfully applied to other strains of C. glutamicum. Both chemicals affected the mycolic acid attachment to the cell surface of C. glutamicum, when INH, the relative percentage of fatty acids of AS019 to the total lipids (mycolic acid plus fatty acids) decreased from 76.9% (in LBG) to 72.9% (in LBG-2% glycine) and 66.4% (in LBG-8 mg InG/ml), thereby suggeting that these chemicals also inhibit fatty acid synthesis.