Proteome analysis of Corynebacterium glutamicum

By the use of different Corynebacterium glutamicum strains move than 1.4 mi llion tons of amino acids, mainly L-glutamate and L-lysine, are produced pe r year. A project was started recently to elucidate the complete DNA sequen ce of this bacterium. In this communication we describe an approach to anal yze the C, glutamicum proteome, based on this genetic information, by a com bination of two-dimensional (2-D) gel electrophoresis and protein identific ation via microsequencing or mass spectrometry. We used these techniques to resolve proteins of C, glutamicum with the aim to establish 2-D protein ma ps as a tool for basic microbiology and for strain improvement. In order to analyze the C. glutamicum proteome, methods were established to fractionat e the C. glutamicum proteins according to functional entities, i.e., cytopl asm, membranes, and cell wall. Protein spots of the cytoplasmic and membran e fraction were identified by N-terminal sequencing, immunodetection, matri x assisted laser desorption/ionization-time of flight-mass spectrometry (MA LDI-TOF-MS) and electrospray ionization-mass spectrometry (ESI-MS). Additio nally, a protocol to analyze proteins secreted by C. glutamicum was establi shed. Approximately 40 protein spots were observed on silver-stained 2-D ge ls, 12 of which were identified.