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Appl Microbiol Biotechnol ; 60(6): 738-42, 2003 Feb.
Article in English | MEDLINE | ID: mdl-12664155

ABSTRACT

A screening procedure was established to identify Corynebacterium glutamicum transposon mutants with an altered L-glutamate excretion behaviour. By this microtiter plate-based approach seven non- or less excreting C. glutamicum strains and two hyper-excreters were found. The subsequently carried out molecular analysis of a hyper-producing clone led to the identification of the gltS gene, which codes for the sodium-coupled secondary L-glutamate uptake system in C. glutamicum. Characterization of a gltS deletion strain revealed that this transporter has a weak but significant impact on L-glutamate production induced by biotin limitation in the wild type. Obviously, GltS leads to the re-uptake of excreted L-glutamate causing a futile cycle. In accord with this hypothesis, the overexpression of gltS decreased L-glutamate production.


Subject(s)
Amino Acid Transport Systems, Acidic/genetics , Bacterial Proteins/genetics , Corynebacterium/genetics , Genes, Bacterial , Glutamic Acid/metabolism , Symporters/genetics , Amino Acid Transport Systems, Acidic/metabolism , Bacterial Proteins/metabolism , Biotin/metabolism , Corynebacterium/metabolism , Escherichia coli/metabolism , Gene Deletion , Gene Expression Regulation, Bacterial , Gene Library , Gene Targeting , Mutagenesis, Insertional , Recombinant Fusion Proteins/metabolism , Sodium/metabolism , Symporters/metabolism
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