Table 2 Enzymatic assay of SpySystem-mediated CYP73A5Δ2-28-ATR2Δ2-77 heterodimers and the free-floating CYP73A5Δ2-28 and ATR2Δ2-77 at increasing concentration of NADPH

From: Tuning architectural organization of eukaryotic P450 system to boost bioproduction in Escherichia coli

Proteins

NADPHa

Km (μM)

kcat (min−1)

kcat/Km (M−1 min−1, × 102)

h

CYP73A5Δ2-28 & ATR2Δ2-77

171.60 ± 7.65

0.041 ± 0.001

2.39

1.68 ± 0.09

heterodimer (I)

235.91 ± 11.29

0.160 ± 0.005

6.78

2.84 ± 0.30

heterodimer (II)

48.56 ± 6.81

0.032 ± 0.001

6.59

1.88 ± 0.58

heterodimer (III)

64.18 ± 2.63

0.008 ± 0.0002

1.25

2.88 ± 0.36

heterodimer (IV)

261.97 ± 21.52

0.681 ± 0.036

26.00

1.97 ± 0.22

  1. aThe Hill’s equation was used to fit the formation rate of p-coumaric acid to the concentrations of NADPH. h is the Hill coefficient. Data are shown as mean ± SE (n = 3 independent experiments).
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