Expression of the Escherichia coli pntAB genes encoding a membrane-bound transhydrogenase in Corynebacterium glutamicum improves L-lysine formation
| Autor: | Armin Kabus, Michael Bott, Tobias Georgi, Volker F. Wendisch |
|---|---|
| Rok vydání: | 2006 |
| Předmět: |
Sucrose
glutamate production Lysine medicine.disease_cause Applied Microbiology and Biotechnology Corynebacterium glutamicum fructose l-glutamate chemistry.chemical_compound 6-bisphosphatase metabolism [NADP] NADP Transhydrogenases glucose bacteria lysine production chemistry.chemical_classification pntAB pntab enzymology [Corynebacterium glutamicum] Escherichia coli Proteins sucrose General Medicine metabolism [NADP Transhydrogenases] growth & development [Corynebacterium glutamicum] Biochemistry central metabolism genetics [Escherichia coli Proteins] Biotechnology enzymology [Escherichia coli] amino-acids cloning Biology metabolism [Escherichia coli Proteins] malic enzyme nicotinamide nucleotide transhydrogenase ddc:570 medicine Escherichia coli methods [Biotechnology] biochemical-characterization genetics [NADP Transhydrogenases] Electrochemical gradient enzymology [Cell Membrane] biosynthesis [Lysine] Cell Membrane Wild type genetics [Escherichia coli] genetics [Corynebacterium glutamicum] Fructose Culture Media Enzyme chemistry fructose-1 NADP |
| Zdroj: | Applied Microbiology and Biotechnology 75, 47-53 (2007). doi:10.1007/s00253-006-0804-9 |
| ISSN: | 0175-7598 |
| DOI: | 10.1007/s00253-006-0804-9 |
| Popis: | A critical factor in the biotechnological production of L: -lysine with Corynebacterium glutamicum is the sufficient supply of NADPH. The membrane-integral nicotinamide nucleotide transhydrogenase PntAB of Escherichia coli can use the electrochemical proton gradient across the cytoplasmic membrane to drive the reduction of NADP(+) via the oxidation of NADH. As C. glutamicum does not possess such an enzyme, we expressed the E. coli pntAB genes in the genetically defined C. glutamicum lysine-producing strain DM1730, resulting in membrane-associated transhydrogenase activity of 0.7 U/mg protein. When cultivated in minimal medium with 10% (w/v) carbon source, the presence of transhydrogenase slightly reduced glucose consumption, whereas the consumption of fructose, glucose plus fructose, and, in particular, sucrose was stimulated. Biomass was increased by pntAB expression between 10 and 30% on all carbon sources tested. Most importantly, the lysine concentration was increased in the presence of transhydrogenase by approximately 10% on glucose, approximately 70% on fructose, approximately 50% on glucose plus fructose, and even by approximately 300% on sucrose. Thus, the presence of a proton-coupled transhydrogenase was shown to be an efficient way to improve lysine production by C. glutamicum. In contrast, pntAB expression had a negative effect on growth and glutamate production of C. glutamicum wild type. |
| Databáze: | OpenAIRE |
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