. 2016 Sep 14;6:33396. doi: 10.1038/srep33396

Table 2. Kinetic parameters of an Asp f3’s and its cysteine mutants’ enzymatic activities, determined by fluorometric and colorimetric assays with t-bOOH and H₂O₂ substrates.

Protein	k_cat, s⁻¹		K_m, M⁻⁶		k_cat/K_m, M⁻¹s⁻¹ × 10⁴
Fluorometric assay (Ampliflu Red)
Asp f3	t-bOOH	H₂O₂	t-bOOH	H₂O₂	t-bOOH	H₂O₂
WT	12.96 ± 0.32	12.53 ± 0.89	70.48 ± 3.12	72.29 ± 5.12	18.39 ± 0.02	17.33 ± 0.12
C31A/C61A	4.92 ± 0.09	3.09 ± 0.08	86.77 ± 1.98	169.50 ± 7.45	5.67 ± 0.01	1.82 ± 0.02
C31S/C61S	1.91 ± 0.12	1.07 ± 0.07	32.01 ± 1.51	57.73 ± 1.65	5.97 ± 0.01	1.85 ± 0.07
Colorimetric assay (NADPH oxidation)
WT	9.92 ± 0.34	10.45 ± 0.91	56.44 ± 6.26	65.04 ± 7.59	17.57 ± 1.34	16.07 ± 0.17
C31A/C61A	3.61 ± 0.09	1.91 ± 0.23	77.20 ± 9.35	148.9 ± 3.72	4.68 ± 0.95	1.28 ± 0.01
C31S/C6 1S	1.81 ± 0.15	1.05 ± 0.07	42.43 ± 2.52	77.20 ± 5.98	4.27 ± 0.89	1.36 ± 0.58

Table 2. Kinetic parameters of an Asp f3’s and its cysteine mutants’ enzymatic activities, determined by fluorometric and colorimetric assays with t-bOOH and H2O2 substrates.