title: Effective sugar nucleotide regeneration for the large-scale enzymatic synthesis of Globo H and SSEA4.
abstract: We report here the development of chemoenzymatic methods for the large-scale synthesis of cancer-associated antigens globopentaose (Gb5), fucosyl-Gb5 (Globo H), and sialyl-Gb5 (SSEA4) by using overexpressed glycosyltransferases coupled with effective regeneration of sugar nucleotides, including UDP-Gal, UDP-GalNAc, GDP-Fuc, and CMP-Neu5Ac. The enzymes used in the synthesis were first identified from different species through comparative studies and then overexpressed in E. coli and isolated for synthesis. These methods provide multigram quantities of products in high yield with only two or three purification steps and are suitable for the evaluation and development of cancer vaccines and therapeutics.
authors.affiliation: Genomics Research Center, Academia Sinica , No. 128, Section 2, Academia Road, Taipei 115, Taiwan.
authors.forename: Tsung-I
authors.initials: TI
authors.lastname: Tsai
authors.forename: Hsin-Yu
authors.initials: HY
authors.lastname: Lee
authors.forename: Shih-Huang
authors.initials: SH
authors.lastname: Chang
authors.forename: Chia-Hung
authors.initials: CH
authors.lastname: Wang
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Tu
authors.forename: Yu-Chen
authors.initials: YC
authors.lastname: Lin
authors.forename: Der-Ren
authors.initials: DR
authors.lastname: Hwang
authors.forename: Chung-Yi
authors.initials: CY
authors.lastname: Wu
authors.forename: Chi-Huey
authors.initials: CH
authors.lastname: Wong
mesh_terms: D015295:Antigens, Tumor-Associated, Carbohydrate / Q000737:chemistry / Q000378:metabolism*; D003001:Cloning, Molecular; D004926:Escherichia coli / Q000235:genetics*; D016695:Glycosyltransferases / Q000235:genetics* / Q000302:isolation & purification / Q000378:metabolism*; D007218:Industrial Microbiology; D015854:Up-Regulation
chemical_list: D015295:Antigens, Tumor-Associated, Carbohydrate; C472765:Globo-H; D016695:Glycosyltransferases