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Subject    One-pot synthesis of GDP-L-fucose by a four-enzyme cascade expressed in Lactococcus lactis
Ling Li, Jin-ho Seo, Nam Soo Han et al.
Journal of Biotechnology
One-pot synthesis of GDP-L-fucose by a four-enzyme cascade expressed in Lactococcus lactis  

Ling Li, Seul-Ah Kim, Ji Eun Heo, Tae-Jip Kim, Jin-Ho Seo, and Nam Soo Han
Journal of Biotechnology
Accept(17.10.16)


Abstract
GDP-L-fucose is an L-fucose donor to synthesize fucosylated compounds such as human milk oligosaccharides or Lewis antigen. In this study, we used Lactococcus lactis subsp. cremoris NZ9000 to express 4 enzymes, ManB, ManC, Gmd, and WcaG and produced GDP-L-fucose by using one-pot synthesis method with mannose-6-phosphate as substrate and the enzymes as biocatalyst. For preparation of enzyme mixture, 4 genes (manB, manC, gmd, and wcaG) cloned from Escherichia coli were transformed into L. lactis strains using pNZ8008 and the recombinant cell lysates were obtained after cultivation. When mannose-6-phosphate was used as the substrate, the consecutive reactions with ManB, ManC, Gmd, and WcaG resulted in the successful production of GDP-L-fucose (0.13 mM). When GDP-D-mannose was used as the substrate, it was entirely converted to GDP-L-fucose (0.2 mM; 0.12 g/L) via 2 enzymatic reactions mediated by Gmd and WcaG. This is the first report of GDP-L-fucose production by using multiple enzymes expressed in lactic acid bacteria.

KEYWORDS : GDP-L-fucose, Lactococcus lactis, mannose-1-phosphate guanylyltransferase, GDP-D-mannose-4,6-dehydratase, GDP-4-keto-6-deoxymannose-3,5-epimerase-4-reductase




No
Category
Subject
Name
Date
Hit
2017     One-pot synthesis of GDP-L-fucose by a four-enzyme cascade expressed in Lactococcus lactis
Ling Li, Jin-ho Seo, Nam Soo Han et al.
Journal of Biotechnology
2017/10/16  328
202 2017     Enhanced production of 2,3-butanediol from xylose by combinatorial engineering of xylose metabolic pathway and cofactor regeneration in pyruvate decarboxylase-deficient Saccharomyces cerevisiae
Soo-Jung Kim,Hee-Jin Sim,Jin-Ho Seo
2017/06/07  797
201 2017     Construction of efficient xylose-fermenting Saccharomyces cerevisiae through a synthetic isozyme system of xylose reductase from Scheffersomyces stipitis
Jung-Hyun Jo, Jin-Ho Seo et al.
Bioresource Technology
2017/05/25  670
200 2017     Metabolic engineering of Saccharomyces cerevisiae for production of spermidine under optimal culture conditions
Sun-Ki Kim, Jung-Hyun Jo, Yong-Cheol Park, Yong-Su Jin, Jin-Ho Seo et al.
Enzyme and Microbial Technology
2017/03/22  1105
199 2017     High production of 2,3-butanediol from glycerol without 1,3-propanediol formation by Raoultella ornithinolytica B6
Taeyeon Kim, Jin-Ho Seo et al.
Applied Microbiology and Biotechnology
2017/03/20  913
198 2016     Metabolic engineering of Saccharomyces cerevisiae for 2,3-butanediol production
Soo-Jung Kim, Jin-Ho Seo. et al.
Applied Microbiology and Biotechnology
2017/02/03  1323
197 2016     Enhanced ethanol fermentation by engineered Saccharomyces cerevisiae strains with high spermidine contents
Sun-Ki Kim, Jung-Hyun Jo, Yong-Su Jin, and Jin-Ho Seo. et al.
Bioprocess and Biosystems Engineering
2016/12/27  1232
196 2016     Bioethanol production from cellulosic hydrolysates by engineered industrial Saccharomyces cerevisiae
Ye-Ji Lee , Yong-Su Jin, Jin-Ho Seo. et al.
Bioresource Technology
2016/12/12  1230
195 2016     Improved production of 2’-fucosyllactose in engineered Escherichia coli by expressing putative α-1,2-fucosyltransferase, WcfB from Bacteroides fragilis
Young-Wook Chin, Jin-Ho Seo. et al.
Journal of Biotechnolog
2016/12/01  1234
194 2016     Enhanced production of 2,3-butanediol by engineered Saccharomyces cerevisiae through fine-tuning of pyruvate decarboxylase and NADH oxidase activities.
Jin-Woo Kim, Jin-Ho Seo. et al.
Biotechnology for Biofuels
2016/11/25  1039
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