title: Improved production of 2'-fucosyllactose in engineered Escherichia coli by expressing putative α-1,2-fucosyltransferase, WcfB from Bacteroides fragilis.
abstract: 2'-Fucosyllactose (2-FL) is one of most abundant oligosaccharides in human milk, which is involved in many biological functions for infant health. Since 2-FL has a great potential in application to functional food materials and pharmaceuticals, several microbial systems for mass production of 2-FL have been developed in recent years. Microbial production of 2-FL was suggested to be influenced by a number of factors including fucosylation activity of α-1,2-fucosyltransferase. In the present study, the wcfB gene coding for α-1,2-fucosyltransferase from Bacteroides fragilis was screened from eleven candidates of putative α-1,2-fucosyltransferase. Introduction of the wcfB gene allows the lacZ-deleted strain of E. coli expressing the genes for guanosine 5'-diphosphate (GDP)-l-fucose biosynthetic enzymes to produce 2-FL. As a result of fed-batch fermentation, 15.4g/L extracellular concentration of 2-FL with 2-FL yield of 0.858g/g lactose and productivity of 0.530g/L/h were obtained. In addition, the feasibility of industrial production of 2-FL using this microbial system was demonstrated by performing fed-batch fermentation in a 75L bioreactor.
authors.affiliation: Research Group of Gut Microbiome, Division of Nutrition and Metabolism Research, Korea Food Research Institute, Seongnam 13539, Republic of Korea.
authors.forename: Young-Wook
authors.initials: YW
authors.lastname: Chin
authors.affiliation: Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Ji-Yeong
authors.initials: JY
authors.lastname: Kim
authors.affiliation: Department of Food and Nutrition, Chungnam National University, Daejeon 34134, Republic of Korea.
authors.forename: Jae-Han
authors.initials: JH
authors.lastname: Kim
authors.affiliation: Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Sang-Min
authors.initials: SM
authors.lastname: Jung
authors.affiliation: Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea. Electronic address: jhseo94@snu.ac.kr.
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
mesh_terms: D001426:Bacterial Proteins / Q000235:genetics; D001441:Bacteroides fragilis / Q000201:enzymology* / Q000235:genetics*; D061249:Batch Cell Culture Techniques; D019149:Bioreactors; D004926:Escherichia coli / Q000235:genetics*; D005285:Fermentation; D005643:Fucose; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial; D005818:Genetic Engineering; D005822:Genetic Vectors; D007763:Lac Operon; D008895:Milk, Human / Q000737:chemistry; D011995:Recombination, Genetic; D017384:Sequence Deletion; D014312:Trisaccharides / Q000096:biosynthesis*
chemical_list: D001426:Bacterial Proteins; D014312:Trisaccharides; D005643:Fucose; D005647:Fucosyltransferases; C031420:2'-fucosyllactose
keywords: 2′-Fucosyllactose; Engineered Escherichia coli; WcfB; α-1,2-Fucosyltransferase