Abstract
Objectives
N-Acetyl-d-neuraminic acid (Neu5Ac) is often synthesized from exogenous N-acetylglucosamine (GlcNAc) and excess pyruvate. We have previously constructed a recombinant Escherichia coli strain for Neu5Ac production using GlcNAc and intracellular phosphoenolpyruvate (PEP) as substrates (Zhu et al. Biotechnol Lett 38:1–9, 2016).
Results
PEP synthesis-related genes, pck and ppsA, were overexpressed within different modes to construct PEP-supply modules, and their effects on Neu5Ac production were investigated. All the PEP-supply modules enhanced Neu5Ac production. For the best module, pCDF-pck-ppsA increased Neu5Ac production to 8.6 ± 0.15 g l−1, compared with 3.6 ± 0.15 g l−1 of the original strain. Neu5Ac production was further increased to 15 ± 0.33 g l−1 in a 1 l fermenter.
Conclusions
The PEP-supply module can improve the intracellular PEP supply and enhance Neu5Ac production, which benefited industrial Neu5Ac production.
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Acknowledgments
This research was supported by the National Natural Science Foundation of China No. 31171640, the Program of Introducing Talents of Discipline to Universities (111-2-06), the Fundamental Research Funds for the Central Universities (JUSRP51504, JUSRP51632A), Key Technology Research and Development Program of Wuxi (CLE01N1208), and Program for the Wuxi Bio-Agriculture Entrepreneurial Leader (130 Plan).
Supporting information
Supplementary Table 1—Strains and plasmids used in this study.
Supplementary Table 2—Primers used in this study.
Supplementary Fig. 1—Time courses of E. coli SA-05/pDTrc-AB harboring various PEP-supply modules.
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Zhu, D., Wu, J., Zhan, X. et al. Phosphoenolpyruvate-supply module in Escherichia coli improves N-acetyl-d-neuraminic acid biocatalysis. Biotechnol Lett 39, 227–234 (2017). https://doi.org/10.1007/s10529-016-2235-8
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DOI: https://doi.org/10.1007/s10529-016-2235-8