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| Hauptseite > Publikationsdatenbank > Toward homosuccinate fermentation: metabolic engineering of Corynebacterium glutamicum for anaerobic production of succinate from glucose and formate |
| Hauptseite > Publikationsdatenbank > Toward homosuccinate fermentation: metabolic engineering of Corynebacterium glutamicum for anaerobic production of succinate from glucose and formate |
| Journal Article | PreJuSER-20662 |
; ;
2012
Soc.
Washington, DC [u.a.]
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Please use a persistent id in citations: doi:10.1128/AEM.07790-11
Abstract: Previous studies have demonstrated the capability of Corynebacterium glutamicum for anaerobic succinate production from glucose under nongrowing conditions. In this work, we have addressed two shortfalls of this process, the formation of significant amounts of by-products and the limitation of the yield by the redox balance. To eliminate acetate formation, a derivative of the type strain ATCC 13032 (strain BOL-1), which lacked all known pathways for acetate and lactate synthesis (Δcat Δpqo Δpta-ackA ΔldhA), was constructed. Chromosomal integration of the pyruvate carboxylase gene pyc(P458S) into BOL-1 resulted in strain BOL-2, which catalyzed fast succinate production from glucose with a yield of 1 mol/mol and showed only little acetate formation. In order to provide additional reducing equivalents derived from the cosubstrate formate, the fdh gene from Mycobacterium vaccae, coding for an NAD(+)-coupled formate dehydrogenase (FDH), was chromosomally integrated into BOL-2, leading to strain BOL-3. In an anaerobic batch process with strain BOL-3, a 20% higher succinate yield from glucose was obtained in the presence of formate. A temporary metabolic blockage of strain BOL-3 was prevented by plasmid-borne overexpression of the glyceraldehyde 3-phosphate dehydrogenase gene gapA. In an anaerobic fed-batch process with glucose and formate, strain BOL-3/pAN6-gap accumulated 1,134 mM succinate in 53 h with an average succinate production rate of 1.59 mmol per g cells (dry weight) (cdw) per h. The succinate yield of 1.67 mol/mol glucose is one of the highest currently described for anaerobic succinate producers and was accompanied by a very low level of by-products (0.10 mol/mol glucose).
Keyword(s): Acetates: metabolism (MeSH) ; Anaerobiosis (MeSH) ; Corynebacterium glutamicum: genetics (MeSH) ; Corynebacterium glutamicum: metabolism (MeSH) ; Fermentation (MeSH) ; Formic Acids: metabolism (MeSH) ; Gene Deletion (MeSH) ; Genes, Bacterial (MeSH) ; Glucose: metabolism (MeSH) ; Lactic Acid: metabolism (MeSH) ; Metabolic Engineering: methods (MeSH) ; Metabolic Networks and Pathways: genetics (MeSH) ; Mutagenesis, Insertional (MeSH) ; Mycobacterium: enzymology (MeSH) ; Mycobacterium: genetics (MeSH) ; Recombination, Genetic (MeSH) ; Succinic Acid: metabolism (MeSH) ; Acetates ; Formic Acids ; Succinic Acid ; Lactic Acid ; Glucose ; formic acid ; J
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