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Molecular cloning of the genes for GDP-mannose 4, 6-dehydratase and GDP-l-fucose synthetase from Bacteroides thetaiotaomicron

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Abstract

Genes encoding GDP-mannose 4, 6-dehydratase (GMD) and GDP-l-fucose synthetase (GFS) were cloned from Bacteroides thetaiotaomicron and overexpressed in recombinant Escherichia coli by constructing isopropyl-β-d-thiogalactopyranoside (IPTG)-inducible expression vectors. GMD and GFS genes from B. thetaiotaomicron were 60 and 45%, respectively, identical to those from E. coli K12 over their entire lengths. An optimum expression condition of 30oC and 0.1 mM IPTG was chosen for maximum soluble expression of B. thetaiotaomicron GMD and GFS in recombinant E. coli BL21(DE3). Functional expression of B. thetaiotaomicron GMD and GFS in recombinant E. coli strains was confirmed by measuring intracellular GDP-l-fucose content.

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References

  1. Boehm G, Stahl B. Oligosaccharides from milk. J. Nutr. 13: 847S–849S (2007)

    Google Scholar 

  2. Kunz C, Rudloff S. Health promoting aspects of milk oligosaccharides. Int. Dairy J. 16: 1341–1346 (2006)

    Article  CAS  Google Scholar 

  3. Newburg DS, Ruiz-Palacios GM, Altaye M, Chaturvedi P, Meinzen-Derr J, Guerrero MD, Morrow AL. Innate protection conferred by fucosylated oligosaccharides of human milk against diarrhea in breastfed infants. Glycobiology 14: 253–263 (2004)

    Article  CAS  Google Scholar 

  4. Kunz C, Rudloff S. Biological function of oligosaccharides in human milk. Acta Paediatr. 82: 903–912 (1993)

    Article  CAS  Google Scholar 

  5. Feizi T, Galustian C. Novel oligosaccharide ligands and ligand-processing pathways for selectins. Trends Biochem. Sci. 24: 369–372 (1999)

    Article  CAS  Google Scholar 

  6. Becker DJ, Lowe JB. Leukocyte adhesion deficiency type II: Review. BBA-Mol. Basis Dis. 1455: 193–204 (1999)

    CAS  Google Scholar 

  7. Kansas GS. Selectins and their ligands: Current concepts and controversies. Blood 88: 3259–3287 (1996)

    CAS  Google Scholar 

  8. Becker DJ, Lowe JB. Fucose: Biosynthesis and biological function in mammals. Glycobiology 13: 41R–53R (2003)

    Article  CAS  Google Scholar 

  9. Yamamoto K, Maruyama T, Kumagai H, Tochikura T, Seno T, Yamaguchi H. Preparation of GDP-l-fucose by using microbial enzymes. Agr. Biol. Chem. Tokyo 48: 823–824 (1984)

    CAS  Google Scholar 

  10. Sullivan FX, Kumar R, Kriz R, Stahl M, Xu GY, Rouse J, Chang XJ, Boodhoo A, Potvin B, Cumming DA. Molecular cloning of human GDP-d-mannose-4, 6-dehydratase and reconstitution of GDP-l-fucose biosynthesis in vitro. J. Biol. Chem. 273: 8193–8202 (1998)

    Article  CAS  Google Scholar 

  11. Albermann C, Distler J, Piepersberg W. Preparative synthesis of GDP-β-l-fucose by recombinant enzymes from enterobacterial sources. Glycobiology 10: 875–881 (2000)

    Article  CAS  Google Scholar 

  12. Comstock LE, Coyne MJ. Bacteroides thetaiotaomicron: A dynamic, niche-adapted human symbiont. Bioessays 25: 926–929 (2003)

    Article  CAS  Google Scholar 

  13. Xu J, Bjursell MK, Himrod J, Deng S, Carmichael LK, Chiang HC, Hooper LV, Gordon JI. A genomic view of the human-Bacteroides thetaiotaomicron symbiosis. Science 299: 2074–2076 (2003)

    Article  CAS  Google Scholar 

  14. Zocco MA, Ainora ME, Gasbarrini G, Gasbarrini A. Bacteroides thetaiotaomicron in the gut: Molecular aspects of their interaction. Digest. Liver Dis. 39: 707–712 (2007)

    Article  CAS  Google Scholar 

  15. Davis GJ, Gloster TM, Henrissat B. Recent structural insights into the expanding world of carbohydrate-active enzymes. Curr. Opin. Struc. Biol. 15: 637–645 (2005)

    Article  Google Scholar 

  16. Webb NA, Mulichak AM, Lam JS, Rocchetta HL, Garavito RM. Crystal structure of a tetrameric GDP-d-mannose 4, 6-dehydratase from a bacterial GDP-d-rhamnose biosynthetic pathway. Protein Sci. 13: 529–539 (2004)

    Article  CAS  Google Scholar 

  17. Thoden JB, Frey PA, Holden HM. Crystal structures of the oxidized and reduced forms of UDP-galactose 4-epimerase isolated from Escherichia coli. Biochemistry 35: 2557–2566 (1996)

    Article  CAS  Google Scholar 

  18. Thoden JB, Wohlers TM, Fridovich-Keil JL, Holden HM. Crystallographic evidence for Tyr 157 functioning as the active site base in human UDP-galactose 4-epimerase. Biochemistry 39: 5691–5701 (2000)

    Article  CAS  Google Scholar 

  19. Somers WS, Stahl ML, Sullivan FX. GDP-fucose synthetase from Escherichia coli: Structure of a unique member of the short-chain dehydrogenase/reductases family that catalyzes two distinct reactions at the same active site. Structure 6: 1601–1612 (1998)

    Article  CAS  Google Scholar 

  20. Allard STM, Beis K, Giraud MF, Hegeman AD, Gross JW, Wilmouth RC, Whitfield C, Graninger M, Messner P, Allen AG, Maskell DJ, Naismith JH. Toward a structural understanding of the dehydratase mechanism. Structure 10: 81–92 (2002)

    Article  CAS  Google Scholar 

  21. Labesse G, Vidal-Cros A, Chomilier J, Gaudry M, Mornon JP. Structural comparisons lead to the definition of a new superfamily of NAD(P)(H)-accepting oxidoreductases: The single-domain reductases/epimerases/dehydrogenases (the ‘RED’ family). Biochem. J. 304: 95–99 (1994)

    CAS  Google Scholar 

  22. Rizzi M, Tonetti M, Vigebani P, Sturla L, Bisso A, Flora AD, Bordo D, Bolognesi M. GDP-4-keto-6-deoxy-d-mannose epimerase/reductase from Escherichia coli, a key enzyme in the biosynthesis of GDP-l-fucose, displays the structural characteristics of the RED protein homology superfamily. Structure 6: 1453–1465 (1998)

    Article  CAS  Google Scholar 

  23. Byun SG, Kim MD, Lee WH, Lee KJ, Han NS, Seo JH. Production of GDP-l-fucose, l-fucose donor for fucosyloligosaccharide synthesis, in recombinant Escherichia coli. Appl. Microbiol. Biot. 74: 768–775 (2007)

    Article  CAS  Google Scholar 

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Authors and Affiliations

  1. School of and Biotechnology and Bioengineering, and Institute of Bioscience and Biotechnology, Kangwon National University, Chuncheon, Gangwon, 200-701, Korea

    Mi-Hee Jang & Myoung-Dong Kim

  2. Department of Agricultural Biotechnology, Seoul National University, Seoul, 151-921, Korea

    Won-Heong Lee, So-Yeon Shin & Jin-Ho Seo

  3. Department of Food Science and Technology, Chungbuk National University, Cheongju, Chungbuk, 361-763, Korea

    Nam Soo Han

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  1. Mi-Hee Jang
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  2. Won-Heong Lee
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  3. So-Yeon Shin
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  4. Nam Soo Han
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  5. Jin-Ho Seo
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  6. Myoung-Dong Kim
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Correspondence to Myoung-Dong Kim.

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Jang, MH., Lee, WH., Shin, SY. et al. Molecular cloning of the genes for GDP-mannose 4, 6-dehydratase and GDP-l-fucose synthetase from Bacteroides thetaiotaomicron . Food Sci Biotechnol 19, 849–855 (2010). https://doi.org/10.1007/s10068-010-0120-0

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  • DOI: https://doi.org/10.1007/s10068-010-0120-0

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