期刊
WORLD JOURNAL OF MICROBIOLOGY & BIOTECHNOLOGY
卷 30, 期 6, 页码 1893-1900出版社
SPRINGER
DOI: 10.1007/s11274-014-1611-6
关键词
Riboflavin; Bacillus subtilis; Gluconeogenesis; Pentose phosphate pathway
资金
- National Program on Key Basic Research Project [2011CBA00804, 2012CB725203]
- National Natural Science Foundation of China [NSFC-21206112, NSFC-21176182]
- National High-tech R&D Program of China [2012AA022103, 2012AA02A702]
- Innovation Foundation of Tianjin University [1308]
The regulation of metabolic flux through glycolytic versus the gluconeogenic pathway plays an important role in central carbon metabolism. In this study, we made an attempt to enhance riboflavin production by deregulating gluconeogenesis in Bacillus subtilis. To this end, gapB (code for NADPH-dependent glyceraldehyde-3-phosphate dehydrogenase), fbp (code for fructose-1,6-bisphosphatase) and pckA (code for phosphoenolpyruvate carboxykinase) were overexpressed in parental strain B. subtilis RH33. Compared with RH33, overexpression of fbp and gapB resulted in approximately 18.0 and 14.2 % increased riboflavin production, respectively, while overexpression of pckA obtained the opposite result. Significant enhancement of riboflavin titers up to 4.89 g/l was obtained in shake flask cultures when gapB and fbp were co-overexpressed, nevertheless the specific growth rate decreased slightly and the specific glucose uptake rate remained almost unchanged. An improvement by 21.9 and 27.8 % of the riboflavin production was achieved by co-overexpression of gapB and fbp in shake flask and fed-batch fermentation, respectively. These results imply that deregulation of gluconeogenesis is an effective strategy for production of metabolites directly stemming from the pentose phosphate pathway as well as other NADPH-demanding compounds with glucose as carbon source in B. subtilis.
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