基于质谱法鉴定碳水化合物异头物构型以确定糖苷水解酶的作用机制。

Mass spectrometry-based identification of carbohydrate anomeric configuration to determine the mechanism of glycoside hydrolases.

作者信息

Shen Yu-Hsuan, Tsai Shang-Ting, Liew Chia Yen, Ni Chi-Kung

机构信息

Institute of Atomic and Molecular Sciences, Academia Sinica, P. O. Box 23-166, Taipei, 10617, Taiwan; Department of Chemistry, National Taiwan University, Taipei, 10617, Taiwan.

Institute of Atomic and Molecular Sciences, Academia Sinica, P. O. Box 23-166, Taipei, 10617, Taiwan.

出版信息

Carbohydr Res. 2019 Apr 1;476:53-59. doi: 10.1016/j.carres.2019.03.004. Epub 2019 Mar 14.

DOI:10.1016/j.carres.2019.03.004

PMID:30897369

Abstract

A rapid mass spectrometry method for determining the anomeric configuration of the sugar at the reducing end of an oligosaccharide was demonstrated. The method was employed to identify the nascent anomeric configuration (i.e., before significant mutarotation occurs) of oligosaccharides released by carbohydrate-active enzymes, which enabled determination of the enzyme mechanism. This method was validated by applying it to various enzymes, including α-glucosidase, β-glucosidases, endoglycoceramidase II, β-galactosidase, and β-amylase.

摘要

展示了一种用于确定寡糖还原端糖的异头构型的快速质谱方法。该方法用于鉴定碳水化合物活性酶释放的寡糖的新生异头构型（即，在显著变旋发生之前），从而能够确定酶的作用机制。通过将该方法应用于多种酶，包括α-葡萄糖苷酶、β-葡萄糖苷酶、内切糖神经酰胺酶II、β-半乳糖苷酶和β-淀粉酶，对该方法进行了验证。