Fig. 2: Designing the parallel cascade. | Nature Communications

Fig. 2: Designing the parallel cascade.

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

Fig. 2

a The upper branch to arylethylamines. i. Dopamine 2 was generated in a one-pot two step cascade reaction from L-tyrosine 4 combining CnTYR and EfTyrDC; ii. Meta-tyramine 10 was produced from meta-tyrosine 9 by EfTyrDC; Reaction conditions i: a 1 mL of reaction mixture (50 mM HEPES, pH 5.5) containing 4 (2.5 mM, 1 equiv.), 0.4 mgmL−1 cell lysate of CnTYR (containing 21% of recombinant protein), 0.4 mgmL−1 cell lysate of EfTyrDC (containing 25% of the recombinant protein), sodium ascorbate 8 (4 equiv.), CuSO4 (40 μM) and PLP (1.25 mM) at 25 °C, 250 rpm for 6 h and quenched by adding 1 μL of trifluoroacetic acid (TFA). ii: a 1 mL of reaction mixture (50 mM HEPES, pH 7.5) containing 7 (2.5 mM, 1 equiv.), 0.4 mgmL−1 cell lysate of EfTyrDC (containing 25% of the recombinant protein), sodium ascorbate 8 (1 equiv.) and PLP (1.25 mM) at 25 °C, 250 rpm for 6 h, and quenched by adding 1 μL of TFA). b The lower branch to the aldehydes. See Table 1 for reaction conditions.

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