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Comparison of the cell surface barrier and enzymatic modification system inBrevibacterium flavum andB. lactofermentum
Authors:Ki-Hyo Jang  Margaret L Britz
Institution:(1) Department of Food and Nutrition, Samcheok National University, 245-711 Gangwon, Korea;(2) School of Agriculture and Food Systems, Gilbert Chandler Campus, The University of Melbourne, Sneydes Rd, 3030 Werribee, Vic, Australia
Abstract:To investigate impediments to plasmid transformation inBrevibacterium flavum BF4 andB. lactofermentum BL1, cell surface barriers were determined by measuring growth inhibition whilst enzymatic barriers were determined by comparing DNA methylation properties.B. lactofermentum was more sensitive to growth inhibition by glycine thanB. flavum. Release of cellular proteins during sonication was more rapid forB. lactofermentum than forB. flavum. Plasmid DNA (pCSL17) isolated fromB. flavum transformed recipient McrBC+ strains ofEscherichia coli with lower efficiency than McrBC.McrBC digestion of this DNA confirmed thatB. flavum contain methylated cytidines in the target sequence ofMcrBC sequences butB. lactofermentum contained a different methylation pattern. DNA derived from theB. lactofermentum transformed recipient EcoKR+ strains ofE. coli with lower efficiency than EcoKR, indicating the presence of methylated adenosines in the target sequence of EcoK sequences. The present data describe the differences in the physical and enzymatic barriers between two species of corynebacteria and also provide some insight into the successful foreign gene expression in corynebacteria.
Keywords:Brevibacterium flavum                      Brevibacterium lactofermentum            identification  transformation
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