Biosynthesis of Ganglioside Oligosaccharide Fluoride in Escherichia coli

doi:10.13560/j.cnki.biotech.bull.1985.2019-0229

Abstract

Abstract:

Gangliosides are glycosphingolipid molecules that present in all vertebrate cells and show important physiological functions and pharmaceutical activities. Synthetic biology provides an opportunity to construct biological synthetic system for the efficient production of gangliosides,in which the synthesis of oligosaccharide fluoride intermediates is a critical step. Herein,the genes of CMP-NeuAc synthase（neuA）from Neisseria meningitidis,α-2,3-sialyltransferase（cst）,β-1,4-GalNAc transferase（cgtA）,β-1,3-galactosyltransferase（cgtB）and UDP-GlcNAc C4 epimerase（gne）from Campylobacter jejuni were introduced into Escherichia coli JM107. Then the biosynthesis pathways of GM3,GM2,and GM1 oligosaccharide fluoride were successfully constructed in E. coli by using lactose fluoride and sialic acid as substrates. A highly sensitive detection method was developed based on pre-column-derivatization-HPLC. The engineered strains were optimized by permeases overexpression and promoter replacement,and the yields of GM3,GM2,and GM1 oligosaccharide fluoride reached 81.5 mg/L,18.1 mg/L,and 12.3 mg/L respectively in shaking flasks fermented,which laid a promising foundation for further transformation by metabolic engineering.

Key words: Escherichia coli, gangliosides, synthetic biology, oligosaccharide fluoride, biosynthesis

LIU Xin-ping, TAN Yu-meng, ZHANG Xue, FENG Yan, YANG Guang-yu. Biosynthesis of Ganglioside Oligosaccharide Fluoride in Escherichia coli[J]. Biotechnology Bulletin, 2019, 35(8): 162-169.

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