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2 results on '"Iris Wiegräbe"'

1. The complete Corynebacterium glutamicum ATCC 13032 genome sequence and its impact on the production of L-aspartate-derived amino acids and vitamins

Author

Brigitte Bathe, Iris Wiegräbe, Alice C. McHardy, Nicole Dusch, Volker F. Wendisch, Jörn Kalinowski, Lothar Eggeling, Bettina Möckel, Reinhard Krämer, Bernhard J. Eikmanns, Lars Gaigalat, Klaus Huthmacher, Andreas Tauch, Oliver Rupp, Michael Hartmann, Daniel Rey, Nicole Bischoff, Daniela Bartels, Alexander Goesmann, Christian Rückert, Folker Meyer, Alfred Pühler, Hermann Sahm, Andreas Burkovski, Walter Pfefferle, Michael Bott, and Burkhard Linke

Subjects
pantothenate overproduction, Proteome, Sequence analysis, Base pair, coryneform bacteria, vitamin production, genome sequence, Molecular Sequence Data, Bioengineering, Biology, Corynebacterium, Applied Microbiology and Biotechnology, Genome, nucleotide-sequence, l-isoleucine, Corynebacterium glutamicum, Multienzyme Complexes, Sequence Homology, Nucleic Acid, Databases, Genetic, biochemical-characterization, phosphotransferase system, molecular analysis, Amino Acid Sequence, Amino Acids, fine-structural analysis, Gene, Peptide sequence, Whole genome sequencing, Genetics, Aspartic Acid, Base Sequence, amino acid production, Gene Expression Profiling, Nucleic acid sequence, bioinformatics, General Medicine, Sequence Analysis, DNA, Vitamins, Recombinant Proteins, Biochemistry, escherichia-coli, bacteria, brevibacterium-flavum, corynebacterium glutamicum, Genome, Bacterial, Biotechnology
Abstract

The complete genomic sequence of Corynebacterium glutainicum ATCC 13032, well-known in industry for the production of amino acids, e.g. of L-glutamate and L-lysine was determined. The C glutamician genome was found to consist of a single circular chromosome comprising 3 282 708 base pairs. Several DNA regions of unusual composition were identified that were potentially acquired by horizontal gene transfer, e.g. a segment of DNA from C diphtheriae and a prophage-containing region. After automated and manual annotation, 3002 protein-coding genes have been identified, and to 2489 of these, functions were assigned by homologies to known proteins. These analyses confirm the taxonomic position of C glutamicum as related to Mycobacteria and show a broad metabolic diversity as expected for a bacterium living in the soil. As an example for biotechnological application the complete genome sequence was used to reconstruct the metabolic flow of carbon into a number of industrially important products derived from the amino acid L-aspartate. (C) 2003 Elsevier B.V. All rights reserved.

Published
2003

2. Development of a Corynebacterium glutamicum DNA microarray and validation by genome-wide expression profiling during growth with propionate as carbon source

Author

Anke Becker, Iris Wiegräbe, Andrea T. Hüser, Michael Dondrup, Iris Brune, Jens Plassmeier, Alfred Pühler, Andreas Tauch, and Jörn Kalinowski

Subjects
genome sequence, Bioengineering, Biology, Corynebacterium, Applied Microbiology and Biotechnology, Corynebacterium glutamicum, Species Specificity, Proline transport, Complementary DNA, Gene expression, Gene cluster, propionate, Oligonucleotide Array Sequence Analysis, chemistry.chemical_classification, Reverse Transcriptase Polymerase Chain Reaction, Gene Expression Profiling, DNA microarray, Chromosome Mapping, General Medicine, Equipment Design, Gene Expression Regulation, Bacterial, Molecular biology, Carbon, Gene expression profiling, Equipment Failure Analysis, 2-methylcitrate cycle, Biochemistry, chemistry, Propionate, Propionates, metabolism, Cell Division, Genome, Bacterial, Biotechnology
Abstract

A DNA microarray was developed to analyse global gene expression of the amino acid-producing bacterium Corynebacterium glutamicum. PCR products representing 93.4% of the predicted C. glutamicum genes were prepared and spotted in quadruplicate onto 3-aminopropyltrimethoxysilane-coated glass slides. The applicability of the C. glutamicum DNA microarray was demonstrated by co-hybridisation with fluorescently labelled cDNA probes. Analysis of the technical variance revealed that C. glutamicum genes detected with different intensities resulting in ratios greater than 1.52 or smaller than -1.52 can be regarded as differentially expressed with a confidence level of greater than 95%. In a validation example, we measured changes of the mRNA levels during growth of C. glutamicum with acetate and propionate as carbon sources. Acetate-grown C glutamicum cultures were used as reference. At the 95% confidence interval, 117 genes revealed increased transcript levels in the presence of propionate, while 43 genes showed a decreased expression compared with the acetate-grown culture. Global expression profiling confirmed the induction of the prpD2B2C2 gene cluster already known to be essential for propionate degradation via the 2-methylcitrate cycle. Besides many genes of unknown function, the paralogous prpD1B1C1 gene cluster as well as fasI-B (encoding fatty-acid synthase IB), dtsR1 and dtsR2 (components of acyl-CoA carboxylases), gluABCD (glutamate transport system), putP (proline transport system), and pyc (pyruvate carboxylase) showed significantly increased expression levels. Differential expression of these genes was confirmed by real-time reverse transcription (RT) PCR assays. (C) 2003 Elsevier B.V. All rights reserved.

Published
2003

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