Biochemistry. 2020 Aug 11;59(31):2903-2908. doi: 10.1021/acs.biochem.0c00390. Epub 2020 Jul 27.
The transglycosylation abilities of β-galactosidases were investigated using hyperpolarized [U-C,U-H]glucose as an acceptor and -nitrophenyl β-galactopyranoside as a donor. Several products were readily observable, and at least in the case when O3 acted as an acceptor, the enzymes showed a clear selectivity toward the β-anomer of glucose. Additionally, it was possible to determine the relative hydrolysis rates of the formed transglycosylation products, providing information on the selectivity as well. Using this method, the transglycosylation abilities of the enzymes could be studied at a very high temporal resolution as well as with high sensitivity, and due to the relative ease of the setup, this method could be more generally applied to investigate glycosidases.
利用超极化 [U-C,U-H]葡萄糖作为受体和对硝基苯-β-半乳糖吡喃糖苷作为供体，研究了β-半乳糖苷酶的转糖苷能力。可以很容易地观察到几种产物，至少在 O3 作为受体的情况下，这些酶对葡萄糖的β-异构体表现出明显的选择性。此外，还可以确定形成的转糖苷产物的相对水解速率，从而提供选择性方面的信息。使用这种方法，可以以非常高的时间分辨率和高灵敏度研究酶的转糖苷能力，并且由于设置相对简单，这种方法可以更广泛地应用于研究糖苷酶。
