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KMID : 0545120010110020294
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Journal of Microbiology and Biotechnology
2001 Volume.11 No. 2 p.294 ~ p.301
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Characterization of the Cell-Surface Barriers to Plasmid Transformation in Corynebacterium glutamicum
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JANG, KI-HYO
CHAMBERS, PAUL J./CHUN, UCK-HAN/BRITZ, MARGARET L.
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Abstract
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The effects of including glycine and isonicotinic acid hydrazide (INH) in the growth medium (Luria broth, LBG) on the subsequent lysozyme-induced protoplast formation and transformation efficiency of Corynebacterium glutamicum were studied. The transformation efficiency of C. glutamicum AS019 increased up to 100-fold as the concentration of glycine in the media increased from 0% to 5% (w/v), relative to cells grown in the absence of glycine. The presence of 5㎎/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 was present at 8㎎/ml in growth media. In the presence of glycine and 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㎎ INH/ml), thereby suggesting that these chemicals also inhibit fatty acid synthesis.
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