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Title

TCA cycle-powered synthesis of fucosylated oligosaccharides.

Authors

Guan, Ningzi; Hyun-Dong Shin; Lingfeng Long; Azadi, Parastoo; Rachel Chen

Abstract

Microbial catalysis has recently emerged as one of the most promising approaches in oligosaccharide synthesis. However, despite significant progress, microbial synthesis still requires much improvement in efficiency and in reduction of process complexity. Additionally, given the stunning diversity and many varied applications of glycans, broadening the range of glycans accessible via microbial synthesis is of paramount importance. Major challenges in microbial synthesis include catabolite repression and high cellular energy requirement. Here we demonstrated a new approach to overcome these challenges by directly tapping into the cellular "power house," the TCA cycle, to provide the cellular energy for synthesis. This approach not only circumvents catabolite repression but also eliminates acidic glycolysis by-products. As such, the whole-cell biocatalysis can be carried out without sophisticated fed-batch feeding and pH control in the synthesis stage. The system could achieve several grams per liter (3-4 g/L) within a 24-h period in shaker flask cultivation for two targets, fucosyllactose and fucosyllactulose, demonstrating efficiency of the biocatalyst developed and its applicability to both natural and non-natural targets. To the best of our knowledge, this is the first use of TCA cycle intermediates as the energy source for oligosaccharide synthesis and the first description of successful synthesis of fucosyllactulose with titers in several grams per liter.

Subjects

MICROBIOLOGICAL synthesis; GLYCOLYSIS; OLIGOSACCHARIDE synthesis; FUCOSYLATION; KREBS cycle

Publication

Glycobiology, 2018, Vol 28, Issue 7, p468

ISSN

0959-6658

Publication type

Academic Journal

DOI

10.1093/glycob/cwy047

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