Table 2.

Fusion activity screening

Enzyme	ADH oxidation k obs (s−1)	BVMO oxidation k obs (s−1)
TmCHMO	–	1.05
ADHA	0.013	–
A-Tm	< 0.01	1.4
Tm-A	< 0.01	1.8
ADHMi	0.11	–
Mi-Tm	< 0.01	0.5
Tm-Mi	0.26	1.1
TbADH	1.35	–
Tb-Tm	1.08	1.9
Tm-Tb	0.28	1.6

Change in absorption at 340 nm measured at 25 °C in 50 mM Tris/HCl pH 8.0. Final substrate concentrations used: 10 mM cyclohexanol and 0.25 mM thioanisole. Cofactor concentration: 100 μM NADP⁺ or NADPH, respectively. For ADHMi and its fusions, 100 μM NAD⁺ was used for the alcohol oxidation. Reaction rates were calculated with protein concentrations determined from absorbance at 441 nm, using the extinction coefficient of TmCHMO (ɛ₄₄₁ = 14.0 mM⁻¹ cm⁻¹)