Efficient synthesis 1,4-cyclohexanedicarboxaldehyde by an engineered alcohol oxidase

Bioresources and Bioprocessing

Table 2 Kinetic constants of AcCO and its variants in production of HMCA

Strains	^aSpecific activity^HMCA (U·g⁻¹)	K_M^HMCA (mM)	k_cat^HMCA (min⁻¹)	^bk_cat/K_M^HMCA (mM⁻¹·min⁻¹)
WT	1.20 ± 0.13	15.30 ± 0.14	0.16 ± 0.028	0.010
W1(S101A)	2.34 ± 0.22	13.45 ± 0.23	0.37 ± 0.064	0.027
W1*(H351V)	2.43 ± 0.22	13.36 ± 0.12	0.41 ± 0.027	0.031
W2(S101A/H351V)	5.31 ± 0.27	10.21 ± 0.46	1.31 ± 0.011	0.13
W3(S101A/H351V/N378S)	6.27 ± 0.15	7.63 ± 0.72	1.87 ± 0.12	0.24
W4(S101A/H351V/N378S/Q329N)	8.76 ± 0.46	1.12 ± 0.19	2.20 ± 0.43	1.96

^aThe specific activity of HMCA was determined with 10 μM purified AcCO (WT) or its variants and 5 mM CHDM in 1 mL air-saturated potassium phosphate buffer (100 mM, pH 8.0) at 30 °C for 30 min
^bThe k_cat/K_M values of HMCA was determined with 10 μM purified AcCO (WT) or its variants and 1–100 mM CHDM in 1 mL air-saturated potassium phosphate buffer (100 mM, pH 8.0) at 30 °C for 30 min