Integration of a Nucleoside Triphosphate Regeneration System in the One‐pot Synthesis of UDP‐sugars and Hyaluronic Acid

Enzymatic synthesis is a promising approach to produce hyaluronic acid (HA) products with defined molecular weight (MW) and dispersity. We here report on the evaluation of nucleoside triphosphate (NTP) regeneration system for precursors uridine-5'-diphosphate α-d-N-acetylglucosamine (UDP-GlcNAc) α-d-glucuronic (UDP-GlcA) integration in one-pot HA. By polyphosphate kinase from Ruegeria pomeroyi (RpPPK2-3) into enzyme cascade synthesize UDP-GlcA we reached 89 % yield after 1 h cycle number (RCN) 17. With UDP-GlcNAc cascade, ATP concentration was lowered 250-fold 94 an RCN 234. Integration RpPPK2-3 HA afforded optimization MgCl2 starting concentrations. 25 mM 0.1 ATP, 0.81 g L−1 average MW 1.17 MDa dispersity about 1.08 75 obtained. conclude that NTP expanded enzymatic possibilities lower range MW.