title: A one-pot approach to bio-synthesize globotriose and its derivatives from simpler substrates.
abstract: Globotriose is involved in numerous pathogenic processes and drug development strategies. Recent studies have demonstrated that globotriosylceramide could be used in colon cancer therapy and as a crucial indicator for susceptibility to HIV-1 infection. Therefore, the cost-effective and facile approaches for large-scale production of globotiose as potential drugs are highly required. Here, a multi-enzyme one-pot system containing a galactokinase (SpGalK, E.C.2.7.1.6), a UDP-glucose pyrophosphorylase (SpGalU, E.C.2.7.7.9), a α-1,4-galactosyltransferase (LgtC, E.C. 2.4.1.44) and a commercial inorganic pyrophosphatase (PPase, EC 3.6.1.1) was designed to achieve globotriose on preparative scales. This method exploits a cheaper initial substrate, galactose, for donor UDP-galactose production. More importantly, the substrate specificity of SpGalK and SpGalU is highly promiscuous and various UDP-galactose derivatives obtained could be used as the donor substrates for LgtC. This pointcut of rapid preparation of globotriose derivatives is proposed for the first time. Finally, three globotriose analogs were achieved by this one-pot multi-enzyme system in our study.
authors.affiliation: The State Key Laboratory of Microbial Technology and School of Life Sciences, National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Xueer
authors.initials: X
authors.lastname: Zhao
authors.affiliation: Hospital for Reproductive Medicine, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Yang
authors.initials: Y
authors.lastname: Zou
authors.affiliation: The State Key Laboratory of Microbial Technology and School of Life Sciences, National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Mengyang
authors.initials: M
authors.lastname: Xue
authors.affiliation: The State Key Laboratory of Microbial Technology and School of Life Sciences, National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Zhongrui
authors.initials: Z
authors.lastname: Ma
authors.affiliation: The State Key Laboratory of Microbial Technology and School of Life Sciences, National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Shuaishuai
authors.initials: S
authors.lastname: Wang
authors.affiliation: The State Key Laboratory of Microbial Technology and School of Life Sciences, National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China.
authors.forename: Peng George
authors.initials: PG
authors.lastname: Wang
authors.affiliation: The State Key Laboratory of Microbial Technology and School of Life Sciences, National Glycoengineering Research Center, Shandong University, Jinan, Shandong 250100, China. Electronic address: chenmin@sdu.edu.cn.
authors.forename: Min
authors.initials: M
authors.lastname: Chen
mesh_terms: D004926:Escherichia coli / Q000235:genetics; D005690:Galactose / Q000378:metabolism*; D014166:Transferases / Q000235:genetics / Q000378:metabolism*; D014312:Trisaccharides / Q000096:biosynthesis* / Q000737:chemistry* / Q000302:isolation & purification
chemical_list: D014312:Trisaccharides; C113460:globotriose; D014166:Transferases; D005690:Galactose
keywords: Derivative; Globotriose; Multi-enzyme; One-pot system