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Methods
Typical procedure for a double stereoselective deracemization with MAO-
N/ATHase (analytical scale). The following stock solutions were prepared.
Catalase from bovine liver (50 kU ml21) was dissolved in MOPS/sodium
formate buffer (0.6 M in MOPS, 3.0 M in sodium formate, pH adjusted with aq.
KOH to pH 7.8). Sav S112T (16.4 mg ml21, 0.75 mM free binding sites, assuming
three free binding sites per tetramer) was dissolved in the catalase containing
buffer. [IrCp*(biot-p-L)Cl] was dissolved in DMF (37.5 mM). Affinity purified
MAO-N-9 (buffer exchange with MOPS, 0.6 M, pH adjusted with KOH to 7.8)
was diluted in MOPS buffer (0.6 M, pH adjusted with KOH to 7.8) to a
concentration of 0.38 mg ml21. Substrate stock was prepared by dissolving the
hydrochloride of 1-ox in H2O (1 M). The ATHase was prepared by adding
[IrCp*(biot-p-L)Cl]-stock (10 ml ml21) to the Sav-stock solution. MAO-N-stock
was placed in 1.5 ml PP-tubes (100 ml) and ATHase was added (100 ml). The
reactions were initiated by addition of substrate stock solution (7.5 ml) and
incubated at 37 8C and 250 r.p.m. in a lying position. For work-up aq. NaOH
solution was added (50 ml of a 10 M solution) and the mixture extracted with
CH2Cl2 (1 × 1 ml). The organic phase was dried over Na2SO4 and analysed by
HPLC (Supplementary Sections S2.3.1 and S2.4.1).
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