Abstract
Corynebacterium glutamicum ATCC13032 and Brevibacterium flavum JV16 were engineered for l-valine production by over-expressing ilvEBN r C genes at 31 °C in 72 h fermentation. Different strategies were carried out to reduce the by-products’ accumulation in l-valine fermentation and also to increase the availability of precursor for l-valine biosynthesis. The native promoter of ilvA of C. glutamicum was replaced with a weak promoter MPilvA (P-ilvAM1CG) to reduce the biosynthetic rate of l-isoleucine. Effect of different relative dissolved oxygen on l-valine production and by-products’ formation was recorded, indicating that 15 % saturation may be the most appropriate relative dissolved oxygen for l-valine fermentation with almost no l-lactic acid and l-glutamate formed. To minimize l-alanine accumulation, alaT and/or avtA was inactivated in C. glutamicum and B. flavum, respectively. Compared to high concentration of l-alanine accumulated by alaT inactivated strains harboring ilvEBN r C genes, l-alanine concentration was reduced to 0.18 g/L by C. glutamicum ATCC13032MPilvA△avtA pDXW-8-ilvEBN r C, and 0.22 g/L by B. flavum JV16avtA::Cm pDXW-8-ilvEBN r C. Meanwhile, l-valine production and conversion efficiency were enhanced to 31.15 g/L and 0.173 g/g by C. glutamicum ATCC13032MPilvA△avtA pDXW-8-ilvEBN r C, 38.82 g/L and 0.252 g/g by B. flavum JV16avtA::Cm pDXW-8-ilvEBN r C. This study provides combined strategies to improve l-valine yield by minimization of by-products’ production.
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This work was financially supported by the Program of Chinese 863 National High-Tech Research and Development Plan Project (No. 2008AA02Z212).
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Hou, X., Chen, X., Zhang, Y. et al. l-Valine production with minimization of by-products’ synthesis in Corynebacterium glutamicum and Brevibacterium flavum . Amino Acids 43, 2301–2311 (2012). https://doi.org/10.1007/s00726-012-1308-9
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DOI: https://doi.org/10.1007/s00726-012-1308-9