¹H NMR analysis of perdeutero N-sulfoheparosan C5-epimerization: a direct way to measure the activity of immobilized C5-epimerase

Abstract

D-Glucuronyl C5-epimerase (C5-epi), which converts D-glucuronic acid to L-iduronic acid, is a crucial enzyme in the preparation of bioengineered heparin, which provides an alternative way to produce safer heparins. Herein, we provided an efficient way to measure the activity of cyanogen bromide-immobilized C5-epi based on proton nuclear magnetic resonance (NMR) analysis of perdeutero N-sulfoheparosan C5-epimerization. In this method, reactions catalyzed by C5-epi could be monitored by real-time 1 D NMR according to the peak intensity change of H-5 from IdoA and GlcA residues in the ¹H NMR spectrum, which is nondestructive of sample and also serves the kinetic property evaluation of immobilized C5-epi.

Graphical Abstract

Keywords:

• ¹H NMR
• C5-epimerase
• immobilized enzyme
• heparan sulfate

Additional information

Funding

This work was supported by grants from National Natural Science Foundation, China [No. 22007049 to X.Z.], Natural Science Foundation of Jiangsu Province [No. BK20200718 to B.L. and BK20200728 to X.Z.].