Journal of Molecular Catalysis B: Enzymatic
Production of deoxy-thymidine-diphosphate (TDP)-sugars as substrates of glycosyltransferases, has been one of main hurdles for combinatorial antibiotic biosynthesis, which combines sugar moiety with aglycon of various antibiotics. Here, we report the one-pot enzymatic synthesis of TDP-2-deoxy-glucose employing high efficient TMP kinase (TMK; E.C. 188.8.131.52), acetate kinase (ACK; E.C. 184.108.40.206), and TDP-glucose synthase (TGS; E.C. 220.127.116.11) with phosphomannomutase (PMM; E.C. 18.104.22.168). In this study, replacing phosphoglucomutase (PGM; E.C. 5.4.2) by PMM from Escherichia coli gave four times higher specific activity on 2-deoxy-6-phosphate glucose, suggesting that the activity on 2-deoxy-glucose-6-phosphate was mainly affected by PMM activity, not PGM activity. Using an in vitro system starting from TMP and 2-deoxy-glucose-6-phosphate glucose, TDP-2-deoxy-glucose (63% yield) was successfully synthesized. Considering low productivity of NDP-sugars from cheap starting materials, this paper showed how production of NDP-sugars could be enhanced by controlling mutase activity.
TDP-2-deoxy-glucose, Phosphomannomutase, Enzymatic synthesis, TMP, One-pot reaction
Yang, Yung-Hum; Kang, Young Bok (Abraham); Kim, Dae-Hee; Lee, Tek-Hyung; Park, Sung-Hee; Lee, Kwangwon; Yoo, Dongwon; Liou, Kwang-kyung; Lee, Hee-Chan; Sohng, Jae-Kyung; and Kim, Byung-Gee, "One-pot Enzymatic Synthesis of Deoxy-thymidine-diphosphate (TDP)-2-deoxy-∝-d-glucose Using Phosphomannomutase" (2010). Faculty Publications - Biomedical, Mechanical, and Civil Engineering. 61.