Fig. 5.

Percent conversions of NDP to NDP–sugar with the OleD variant Loki. Reactions contained 0.5 mM of 2-chloro-4-nitrophenyl glycoside donor (donors 1 and 4–8), 1.0 mM NDP, and 5.25 μM (50 μg) of purified Loki variant in Tris⋅HCl buffer (50 mM, pH 8.0) and a final volume of 200 μL. Absorbance at 410 nm was followed and reactions were halted when they reached completion, increase in absorbance halted, or total reaction time reached 24 h (SI Appendix). Percent conversions were determined by HPLC. The SD for production of UDP–α-D-glucose (donor 9) across multiple runs (n = 3) was <10%.