Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지
DOI QR코드

DOI QR Code

A Simple Enzymatic Method for Quantitation of 2'-Fucosyllactose

  • Seydametova, Emine (Department of Integrative Biotechnology, College of Biotechnology and Bioengineering, Sungkyunkwan University) ;
  • Shin, Jonghyeok (Department of Integrative Biotechnology, College of Biotechnology and Bioengineering, Sungkyunkwan University) ;
  • Yu, Jiwon (Department of Integrative Biotechnology, College of Biotechnology and Bioengineering, Sungkyunkwan University) ;
  • Kweon, Dae-Hyuk (Department of Integrative Biotechnology, College of Biotechnology and Bioengineering, Sungkyunkwan University)
  • Received : 2018.03.07
  • Accepted : 2018.05.01
  • Published : 2018.07.28
Download PDF
⟨ Previous Next ⟩

Abstract

2'-Fucosyllactose (2'-FL) is one of the most important human milk oligosaccharides and has several health benefits for infants. The levels of 2'-FL in breast milk or samples from other sources can be quantified by high-performance liquid chromatography. However, this method cannot be used for simultaneous detection of the target compound in numerous samples. Here, we developed a simple method for quantifying 2'-FL in a microplate format. The method involves two steps: (i) release of $\text\tiny{L}$-fucose from 2'-FL by ${\alpha}$-(1-2,3,4,6)-$\text\tiny{L}$-fucosidase and (ii) measurement of NADPH formed during the oxidation of $\text\tiny{L}$-fucose by $\text\tiny{L}$-fucose dehydrogenase. This method enables measurement of up to 5 g/l 2'-FL in 50 min using a 96-well microplate. The efficiency and simplicity of the proposed method make it suitable for the analyses of a large number of samples simultaneously.

Keywords

References

  1. Petschacher B, Nidetzky B. 2016. Biotechnological production of fucosylated human milk oligosaccharides: prokaryotic fucosyltransferases and their use in biocatalytic cascades or whole cell conversion systems. J. Biotechnol. 235: 61-83. https://doi.org/10.1016/j.jbiotec.2016.03.052
  2. Castanys-Munoz E, Martin MJ, Prieto PA. 2013. 2'- Fucosyllactose: an abundant, genetically determined soluble glycan present in human milk. Nutr. Rev. 71: 773-789. https://doi.org/10.1111/nure.12079
  3. Barile D, Rastall RA. 2013. Human milk and related oligosaccharides as prebiotics. Curr. Opin. Biotechnol. 24: 214-219. https://doi.org/10.1016/j.copbio.2013.01.008
  4. Morrow AL, Ruiz-Palacios G, Altaye M, Jiang X, Guerrero ML, Meinzen-Derr JK, et al. 2004. Human milk oligosaccharides are associated with protection against diarrhea in breast-fed infants. J. Pediatr. 145: 297-303. https://doi.org/10.1016/j.jpeds.2004.04.054
  5. Newburg DS, Ruiz-Palacios GM, Morrow AL. 2005. Human milk glycans protect infants against enteric pathogens. Annu. Rev. Nutr. 25: 37-58. https://doi.org/10.1146/annurev.nutr.25.050304.092553
  6. Magalhaes A, Reis CA. 2010. Helicobacter pylori adhesion to gastric epithelial cells is mediated by glycan receptors. Braz. J. Med. Biol. Res. 43: 611-618. https://doi.org/10.1590/S0100-879X2010007500049
  7. Weichert S, Jennewein S, Hufner E, Weiss C, Borkowski J, Putze J, et al. 2013. Bioengineered 2'-fucosyllactose and 3'- fucosyllactose inhibit the adhesion of Pseudomonas aeruginosa and enteric pathogens to human intestinal and respiratory cell lines. Nutr. Res. 33: 831-838. https://doi.org/10.1016/j.nutres.2013.07.009
  8. Gonia S, Tuepker M, Heisel T, Autran C, Bode L, Gale CA. 2015. Human milk oligosaccharides inhibit Candida albicans invasion of human premature intestinal epithelial cells. J. Nutr. 145: 1992-1998. https://doi.org/10.3945/jn.115.214940
  9. Kunz C, Rudloff S, Baier W, Klein N, Strobel S. 2000. Oligosaccharides in human milk: structural, functional, and metabolic aspects. Annu. Rev. Nutr. 20: 699-722. https://doi.org/10.1146/annurev.nutr.20.1.699
  10. Chaturvedi P, Warren CD, Altaye M, Morrow AL, Ruiz- Palacios G, Pickering LK, et al. 2001. Fucosylated human milk oligosaccharides vary between individuals and over the course of lactation. Glycobiology 11: 365-372. https://doi.org/10.1093/glycob/11.5.365
  11. Lee WH, Pathanibul P, Quarterman J, Jo JH, Han NS, Miller MJ, et al. 2012. Whole cell biosynthesis of a functional oligosaccharide, 2'-fucosyllactose, using engineered Escherichia coli. Microb. Cell Fact. 11: 48. https://doi.org/10.1186/1475-2859-11-48
  12. Thurl S, Muller-Verner B, Sawatzki G. 1996. Quantification of individual oligosaccharide compounds from human milk using high-pH anion-exchange chromatography. Anal. Biochem. 235: 202-206. https://doi.org/10.1006/abio.1996.0113
  13. Chaturvedi P, Warren CD, Ruiz-Palacios G, Pickering LK, Newburg DS. 1997. Milk oligosaccharide profiles by reversedphase HPLC of their perbenzoylated derivatives. Anal. Biochem. 251: 89-97. https://doi.org/10.1006/abio.1997.2250
  14. Bao Y, Chen C, Newburg DS. 2013. Quantification of neutral human milk oligosaccharides by graphitic carbon highperformance liquid chromatography with tandem mass spectrometry. Anal. Biochem. 433: 28-35. https://doi.org/10.1016/j.ab.2012.10.003
  15. Chin YW, Seo N, Kim JH, Seo JH. 2016. Metabolic engineering of Escherichia coli to produce 2'-fucosyllactose via salvage pathway of guanosine 5'-diphosphate (GDP)-Lfucose. Biotechnol. Bioeng. 113: 2443-2452. https://doi.org/10.1002/bit.26015
  16. Becker DJ, Lowe JB. 2003. Fucose: biosynthesis and biological function in mammals. Glycobiology 13: 41R-53R. https://doi.org/10.1093/glycob/cwg054
  17. Berteau O, McCort I, Goasdoue N, Tissot B, Daniel R. 2002. Characterization of a new alpha-L-fucosidase isolated from the marine mollusk Pecten maximus that catalyzes the hydrolysis of alpha-L-fucose from algal fucoidan (Ascophyllum nodosum). Glycobiology 12: 273-282. https://doi.org/10.1093/glycob/12.4.273
  18. Sakurama H, Tsutsumi E, Ashida H, Katayama T, Yamamoto K, Kumagai H. 2012. Differences in the substrate specificities and active-site structures of two alpha-Lfucosidases (glycoside hydrolase family 29) from Bacteroides thetaiotaomicron. Biosci. Biotechnol. Biochem. 76: 1022-1024. https://doi.org/10.1271/bbb.111004
  19. Aminoff D, Furukawa K. 1970. Enzymes that destroy blood group specificity. I. Purification and properties of ${\alpha}$-Lfucosidase from Clostridium perfringens. J. Biol. Chem. 245: 1659-1669.
  20. Bahl OP. 1970. Glycosidases of Aspergillus niger. II. Purification and general properties of 1,2-alpha-L-fucosidase. J. Biol. Chem. 245: 299-304.
  21. Kochibe N. 1973. Purification and properties of ${\alpha}$-L-fucosidase from Bacillus fulminans. J. Biochem. 74: 1141-1149. https://doi.org/10.1093/oxfordjournals.jbchem.a130341
  22. Zivkovic AM, German JB, Lebrilla CB, Mills DA. 2011. Human milk glycobiome and its impact on the infant gastrointestinal microbiota. Proc. Natl. Acad. Sci. USA 108: 4653-4568. https://doi.org/10.1073/pnas.1000083107
  23. Chin YW, Kim JY, Lee WH, Seo JH. 2015. Enhanced production of 2'-fucosyllactose in engineered Escherichia coli BL21star(DE3) by modulation of lactose metabolism and fucosyltransferase. J. Biotechnol. 210: 107-115. https://doi.org/10.1016/j.jbiotec.2015.06.431

Cited by

  1. Development of fluorescent Escherichia coli for a whole-cell sensor of 2ʹ-fucosyllactose vol.10, pp.None, 2020, https://doi.org/10.1038/s41598-020-67359-x