Table 4 Parallel cascades with CnTYR variants, EfTyrDC, CvTAm, and TfNCS-A79I to halogenated BIAs

From: Enzymatic synthesis of benzylisoquinoline alkaloids using a parallel cascade strategy and tyrosinase variants

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  1. aYields were determined by HPLC analysis at 280 nm against products standards. bFor preparative-scale reactions products were purified by preparative HPLC or an extraction method. cees were determined by chiral HPLC. Reaction conditions: Details are specific to each cascade. E.g., entry 4, 50 mL reaction mixture A (RMA) - HEPES (20 mM, pH 5.0), 11 (10 mM, 1 equiv,), CnTYR-M1 (N201S/G205R/V206I) and EfTyrDC (10% (v/v) cell lysates), 8 (4 equiv.), PLP (0.5 equiv.), CuSO4 (5 μM) was stirred at 25 °C for 8 h. A 20 mL reaction mixture B (RMB) - HEPES (50 mM, pH 7.5), 11 (20 mM, 1 equiv.), EfTyrDC and CvTAm (10% and 5% (v/v) cell lysates, respectively), 8 (4 equiv.), PLP (1 equiv.), 15 (1 equiv.) was stirred at 37 °C for 8 h. Afterwards, RMA and RMB were combined, TfNCS-A79I (10% (v/v) cell lysates) added, and the reaction stirred at 37 °C for 16 h.
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