Extended Data Fig. 3: Optimization of the in vitro reaction conditions for ApPT-catalyzed oxidation of 1.
a, Reaction parameters optimization including alternative redox partners (RPs), biocatalyst loads, as well as the pH. Yields were determined by HPLC analysis. Conditions: 2 mM 1 in 5% (v/v) DMSO, 1 mM NADP+, 100 mM Na2HPO3, opt13, and ApPT/RPs in 100 mM kpi buffer at 23 °C, 220 rpm for 20 h. aBiocatalyst loads in entries 1–6 can be effectively determined according to the Supplementary Methods. Due to the biocatalysts in entries 7–10 cannot be purified, the hydroxylated activity was determined using the same amount of crude enzyme lysate with OD600 = 30. b, Different types and expression modes of RPs with ApPT. RP1 and RP2 represent self-sufficient ApPT chimeras that were expressed in the pET28a(+) plasmid. RP3, RP4, and RP5 represent RPs that were individually expressed alongside ApPT in the pRSFDuet-1 plasmid.
