Table 1 Michaelis–Menten steady-state parameters of CALB on PET derivatives

Substrate	pH	K_M (mM)	V_max (U/mg)	k_cat (s⁻¹)	k_cat / K_M (mM⁻¹ x s⁻¹)	\(\triangle {G}_{\exp }^{{act}}\)
BHET	5	22.5^a ± 9.6	0.94 ± 0.14	0.52^a ± 0.07	0.023^b,c ± 0.010	17.8
	7	13.3 ± 4.3	1.62 ± 0.27	0.89 ± 0.15	0.067^d ± 0.024	17.5
	9	61.0^a ± 25.7	1.33 ± 0.45	0.73^a ± 0.25	0.012^c,e ± 0.006	17.6
MHET	5	23.8^a ± 8.8	2.27 ± 0.53	1.25^a ± 0.29	0.052^b,d ± 0.023	17.3
	7	15.1 ± 3.2	0.25 ± 0.02	0.14 ± 0.011	0.009^e ± 0.002	18.6
	9	>100	n.a.	n.a.	n.a.	na

Activity assay: Substrate in 10% DMSO and 100 mM buffer at 25 °C. Buffer for pH 5 was sodium acetate, for pH 7 was sodium phosphate and for pH 9 was sodium bicarbonate. n.a. not assessed because it did not reach substrate saturation. ^aValues were extrapolated out of the upper limit of 20 and 50 mM of BHET and MHET as higher concentrations cannot be achieved under the activity assay conditions due to the solubility limit of both substrates.^b,c,d,e Means with a different letter are significantly different (p < 0.04124). Data were determined from three independent replicates and analyzed by ordinary one-way ANOVA with Tukey’s multiple comparison test.

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