title: Metabolic engineering of Escherichia coli to produce 2'-fucosyllactose via salvage pathway of guanosine 5'-diphosphate (GDP)-l-fucose.
abstract: 2'-Fucosyllactose (2-FL) is one of the key oligosaccharides in human milk. In the present study, the salvage guanosine 5'-diphosphate (GDP)-l-fucose biosynthetic pathway from fucose was employed in engineered Escherichia coli BL21star(DE3) for efficient production of 2-FL. Introduction of the fkp gene coding for fucokinase/GDP-l-fucose pyrophosphorylase (Fkp) from Bacteroides fragilis and the fucT2 gene encoding α-1,2-fucosyltransferase from Helicobacter pylori allows the engineered E. coli to produce 2-FL from fucose, lactose and glycerol. To enhance the lactose flux to 2-FL production, the attenuated, and deleted mutants of β-galactosidase were employed. Moreover, the 2-FL yield and productivity were further improved by deletion of the fucI-fucK gene cluster coding for fucose isomerase (FucI) and fuculose kinase (FucK). Finally, fed-batch fermentation of engineered E. coli BL21star(DE3) deleting lacZ and fucI-fucK, and expressing fkp and fucT2 resulted in 23.1 g/L of extracellular concentration of 2-FL and 0.39 g/L/h productivity. Biotechnol. Bioeng. 2016;113: 2443-2452. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Department of Agricultural Biotechnology and Center for Food and Bioconvergence, Seoul National University, Seoul 08826, Republic of Korea.
authors.forename: Young-Wook
authors.initials: YW
authors.lastname: Chin
authors.affiliation: Graduate School of Analytical Science and Technology, Chungnam National University, Daejeon, Republic of Korea.
authors.forename: Nari
authors.initials: N
authors.lastname: Seo
authors.affiliation: Department of Food and Nutrition, Chungnam National University, Daejeon, 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. jhseo94@snu.ac.kr.
authors.forename: Jin-Ho
authors.initials: JH
authors.lastname: Seo
mesh_terms: D004926:Escherichia coli / Q000502:physiology*; D005647:Fucosyltransferases / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000502:physiology; D006154:Guanosine Diphosphate Fucose / Q000235:genetics / Q000378:metabolism*; D007785:Lactose / Q000378:metabolism; D060847:Metabolic Engineering / Q000379:methods*; D015398:Signal Transduction / Q000502:physiology; D014312:Trisaccharides / Q000096:biosynthesis* / Q000235:genetics
chemical_list: D014312:Trisaccharides; D006154:Guanosine Diphosphate Fucose; D005647:Fucosyltransferases; D007785:Lactose; C031420:2'-fucosyllactose
keywords: 2′-fucosyllactose; engineered Escherichia coli; fucokinase/GDP-l-fucose pyrophosphorylase; fucose