Modulation of guanosine 5′-diphosphate-d
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- 作者：Won-Heong Lee ; Nam-Soo Han ; Yong-Cheol Park ; Jin-Ho Seo
- 关键词：Recombinant Escherichia coli ; GDP-l-fucose ; GDP-d-mannose ; Fed-batch fermentation
- 刊名：Bioresource Technology
- 出版年：2009
- 出版时间：December 2009
- 年：2009
- 卷：100
- 期：24
- 页码：6143-6148
- 全文大小：546 K
文摘
Guanosine 5’-diphosphate (GDP)-l-fucose, an activated form of a nucleotide sugar, plays an important role in a wide range of biological functions. In this study, the enhancement of GDP-l-fucose production was attempted by supplementation of mannose, which is a potentially better carbon source to be converted into GDP-l-fucose than glucose, and combinatorial overexpression of the genes involved in the biosynthesis of GDP-d-mannose, a precursor of GDP-l-fucose. Supply of a mannose and glucose led to a 1.3-fold-increase in GDP-l-fucose concentration (52.5 ± 0.8 mg l−1) in a fed-batch fermentation of recombinant E. coli BL21star(DE3) overexpressing the gmd and wcaG genes, compared with the case using glucose as a sole carbon source. A maximum GDP-l-fucose concentration of 170.3 ± 2.3 mg l−1, corresponding to a 4.4-fold enhancement compared with the control strain overexpressing gmd and wcaG genes only, was achieved in a glucose-limited fed-batch fermentation of a recombinant E. coli BL21star(DE3) strain overexpressing manB, manC, gmd and wcaG genes. Further improvement of GDP-l-fucose production was not obtained by additional overexpression of the manA gene.