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Proteome analysis of Corynebacterium glutamicum
Author(s) -
Hermann Thomas,
Pfefferle Walter,
Baumann Christian,
Busker Eberhard,
Schaffer Steffen,
Bott Michael,
Sahm Hermann,
Dusch Nicole,
Kalinowski Jörn,
Pühler Alfred,
Bendt Anne K.,
Krämer Reinhard,
Burkovski Andreas
Publication year - 2001
Publication title -
electrophoresis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.666
H-Index - 158
eISSN - 1522-2683
pISSN - 0173-0835
DOI - 10.1002/1522-2683(200105)22:9<1712::aid-elps1712>3.0.co;2-g
Subject(s) - corynebacterium glutamicum , proteome , mass spectrometry , tandem mass spectrometry , chemistry , lysine , biochemistry , bacteria , proteomics , electrospray ionization , chromatography , amino acid , biology , gene , genetics
By the use of different Corynebacterium glutamicum strains more than 1.4 million tons of amino acids, mainly L ‐glutamate and L ‐lysine, are produced per year. A project was started recently to elucidate the complete DNA sequence of this bacterium. In this communication we describe an approach to analyze the C. glutamicum proteome, based on this genetic information, by a combination of two‐dimensional (2‐D) gel electrophoresis and protein identification via microsequencing or mass spectrometry. We used these techniques to resolve proteins of C. glutamicum with the aim to establish 2‐D protein maps as a tool for basic microbiology and for strain improvement. In order to analyze the C. glutamicum proteome, methods were established to fractionate the C. glutamicum proteins according to functional entities, i.e. , cytoplasm, membranes, and cell wall. Protein spots of the cytoplasmic and membrane fraction were identified by N ‐terminal sequencing, immunodetection, matrix assisted laser desorption/ionization‐time of flight‐mass spectrometry (MALDI‐TOF‐MS) and electrospray ionization‐mass spectrometry (ESI‐MS). Additionally, a protocol to analyze proteins secreted by C. glutamicum was established. Approximately 40 protein spots were observed on silver‐stained 2‐D gels, 12 of which were identified.