Effective Sugar Nucleotide Regeneration for the Large-Scale Enzymatic Synthesis of Globo H and SSEA4

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• 作者：Tsung-I Tsai ; Hsin-Yu Lee ; Shih-Huang Chang ; Chia-Hung Wang ; Yu-Chen Tu ; Yu-Chen Lin ; Der-Ren Hwang ; Chung-Yi Wu ; Chi-Huey Wong
• 刊名：Journal of the American Chemical Society
• 出版年：2013
• 出版时间：October 2, 2013
• 年：2013
• 卷：135
• 期：39
• 页码：14831-14839
• 全文大小：344K
• 年卷期：v.135,no.39(October 2, 2013)
• ISSN：1520-5126

文摘

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.