Enhanced N-acetyl-d-neuraminic production from glycerol and N-acetyl-d-glucosamine by metabolically engineered Escherichia coli with a two-stage pH-shift control strategy
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Typical N-acetyl-d-neuraminic acid (Neu5Ac) production uses N-acetyl-d-glucosamine (GlcNAc) and excess pyruvate as substrates in the enzymatic or whole-cell biocatalysis process. In a previous study, a Neu5Ac-producing biocatalytic process via engineered Escherichia coli SA-05/pDTrc-AB/pCDF-pck-ppsA was constructed without exogenous pyruvate. In this study, glycerol was found to be a good energy source compared with glucose for the catalytic system with resting cells, and Neu5Ac production increased to 13.97 ± 0.27 g L−1. In addition, a two-stage pH shift strategy was carried out, and the Neu5Ac yield was improved to 14.61 ± 0.31 g L−1. The GlcNAc concentration for Neu5Ac production was optimized. Finally, an integrated strategy was developed for Neu5Ac production, and the Neu5Ac yield reached as high as 18.17 ± 0.27 g L−1. These results provide a new biocatalysis technology for Neu5Ac production without exogenous pyruvate.
KeywordsBiocatalytic process Fed-batch N-Acetyl-d-neuraminic acid Glycerol Energy source
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), and the National first-class discipline program of light industry technology and engineering (LITE2018-17).
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