. 2020 Mar 23;21(6):2208. doi: 10.3390/ijms21062208

Table 2.

Comparison of kinetic values of purified UGT72B27 enzyme obtained via three different detection methods. (a) pH-sensitive assay, data taken from [35], obtained by single measurements, (b) UDP-Glo^TM assay, measured in 384-well plate, and (c) phosphate glycosyltransferase assay, measured in 96-well plate. Substrate concentrations were varied from 10–3000 μM. Guaiacol: 2-Methoxyphenol, DMP: 2,6-Dimethoxyphenol, MMP: 2-Methoxy-4-methylphenol, MDMP: 4-Methyl-2,6-dimethoxyphenol. n = 3. (*) since the publication, this value was corrected.

Substrate	a. pH-Sensitive Assay			b. UDP-Glo^TM Assay			c. Phosphate GT Assay
Substrate	K_M (µM)	k_cat (s⁻¹)	k_cat/K_M (mM⁻¹ s⁻¹)	K_M (µM)	k_cat (s⁻¹)	k_cat/K_M (mM⁻¹ s⁻¹)	K_M (µM)	k_cat (s⁻¹)	k_cat/K_M (mM⁻¹ s⁻¹)
Guaiacol	*32 `±` 1	2.3 `±` 0.02	72.5 `±` 3	23 ± 1	0.08 `±` 0.02	3.7 ± 1	28 `±` 3	0.13 `±` 0.003	4.7 ± 0.6
trans-Resveratrol	36 `±` 5	0.6 `±` 0.05	17.0 ± 3.8	21 `±` 5	0.004 ± 0.0001	0.2 ± 0.05	15 `±` 1	0.02 `±` 0.002	1.3 ± 0.2
Thymol	53 `±` 0.25	0.7 `±` 0.07	13.5 ± 1.4	28 `±` 7	0.04 `±` 0.003	1.4 `±` 0.1	20 `±` 1	0.07 `±` 0.001	3.3 ± 0.2
DMP	211 `±` 52	1.9 `±` 0.05	8.8 ± 2.4	23 `±` 3	0.09 `±` 0.003	3.7 ± 0.6	41 `±` 4	0.1 `±` 0.004	2.3 ± 0.3
MMP	115 ± 22	0.9 `±` 0.01	8.0 ± 1.6	41 `±` 9	0.06 `±` 0.001	1.4 ± 0.3
m-Cresol	48 `±` 19	0.4 `±` 0.02	7.9 ± 3.5	14 `±` 3	0.04 `±` 0.002	2.6 ± 0.6	15 `±` 2	0.05 `±` 0.0005	3.4 ± 0.5
Phloroglucinol	77 `±` 10	0.5 `±` 0.05	7.1 ± 1.6	35 `±` 4	0.05 `±` 0.006	1.3 ± 0.3	47 `±` 4	0.09 `±` 0.006	1.9 ± 0.3
o-Cresol	148 `±` 14	0.5 `±` 0.04	3.6 ± 0.6	40 `±` 5	0.05 `±` 0.002	1.4 ± 0.2	32 `±` 2	0.06 `±` 0.005	1.8 ± 0.3
MDMP	278 `±` 22	0.5 `±` 0.04	1.9 ± 0.3	173 ± 28	0.05 `±` 0.005	0.3 ± 0.07	143 `±` 13	0.06 `±` 0.002	0.5 ± 0.1
Phenol	326 `±` 83	0.6 `±` 0.04	1.8 ± 0.6	153 `±` 24	0.04 `±` 0.002	0.3 ± 0.06	62 `±` 0.5	0.07 `±` 0.007	1.1 ± 0.1
Furaneol	478 `±` 45	0.5 `±` 0.04	1.0 `±` 0.2	453 `±` 35	0.02 `±` 0.002	0.04 ± 0.007