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Enhancement of riboflavin production by deregulating gluconeogenesis in Bacillus subtilis

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Abstract

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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Abbreviations

PPP:

Pentose phosphate pathway

Ru5P:

Ribulose-5P

FBPase:

Fructose 1,6-bisphosphatase

MM:

Minimal medium

CDW:

Cell dry weight

TCA:

Tricarboxylic acid

PEP:

Phosphoenolpyruvate

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Acknowledgments

We thank Dr. Zhenquan Lin for the help of qRT-PCR analysis. This work was supported by 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) and the Innovation Foundation of Tianjin University (1308).

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Authors and Affiliations

  1. Department of Biochemical Engineering, School of Chemical Engineering and Technology, Tianjin University, 92# Weijin Road, Nankai District, Tianjin, 300072, People’s Republic of China

    Guanglu Wang, Ling Bai, Zhiwen Wang, Ting Shi, Tao Chen & Xueming Zhao

  2. Key Laboratory of Systems Bioengineering, Ministry of Education, Tianjin University, Tianjin, 300072, People’s Republic of China

    Guanglu Wang, Ling Bai, Zhiwen Wang, Ting Shi, Tao Chen & Xueming Zhao

  3. Collaborative Innovation Center of Chemical Science and Engineering (Tianjin), Tianjin University, Tianjin, 300072, People’s Republic of China

    Guanglu Wang, Ling Bai, Zhiwen Wang, Ting Shi, Tao Chen & Xueming Zhao

  4. Edinburgh-Tianjin Joint Research Centre for Systems Biology and Synthetic Biology, Tianjin University, Tianjin, 300072, People’s Republic of China

    Guanglu Wang, Ling Bai, Zhiwen Wang, Ting Shi, Tao Chen & Xueming Zhao

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  1. Guanglu Wang
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  2. Ling Bai
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  3. Zhiwen Wang
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  4. Ting Shi
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Correspondence to Zhiwen Wang.

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Guanglu Wang and Ling Bai have contributed equally to this work.

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Wang, G., Bai, L., Wang, Z. et al. Enhancement of riboflavin production by deregulating gluconeogenesis in Bacillus subtilis . World J Microbiol Biotechnol 30, 1893–1900 (2014). https://doi.org/10.1007/s11274-014-1611-6

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  • DOI: https://doi.org/10.1007/s11274-014-1611-6

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