TABLE 2.

Substrate specificities of chloromuconate cycloisomerase and MCI from different strains and of trans-DLH from Pseudomonas sp. strain MT1

Strain	Enzyme	Substrate	Activity with 0.1 mM substrate (U/mg)a	Km (μM)b	kcat (min−1)b	kcat/Km (min−1 μM−1)b
Pseudomonas sp. strain MT1	MCI	Muconate	9.9 ± 1.2	1,730 ± 350	7,500 ± 350	4.3
		3-Chloromuconate	35 ± 6	410 ± 75	7,200 ± 520	17.6
		2-Chloromuconate	0.09 ± 0.02	NDc	ND	ND
		2,4-Dichloromuconate	<0.001	ND	ND	ND
Pseudomonas sp. strain MT1	trans-DLH	trans-Dienelactone	710 ± 37	480 ± 20	159,000 ± 8,500	330
		4-Fluoromuconolactone	190 ± 4	1,200 ± 20	98,000 ± 2,200	82
R. eutropha JMP 134	CMCI (= TfdDI)d	Muconate	1.6	996 ± 35	700 ± 10	0.71
		3-Chloromuconate	48	98 ± 3	3,770 ± 70	38
		2-Chloromuconate	3	79 ± 3	215 ± 5	2.7
		2,4-Dichloromuconate	57.5	23 ± 1	2,810 ± 40	120
Pseudomonas sp. strain B13	CMCI	Muconate	3.3	270 ± 55	480 ± 70	1.8
		3-Chloromuconate	21.7	160 ± 55	2,240 ± 480	14
		2-Chloromuconate	8.5	42 ± 6	480 ± 30	11
		2,4-Dichloromuconate	7.9	26 ± 2	390 ± 10	15
P. putida PRS2000	MCI	Muconate	223	40 ± 4	12,600 ± 400	310
		3-Chloromuconate	0.38	320 ± 200	64 ± 30	0.20
		2-Chloromuconate	0.76	59 ± 5	49 ± 2	0.83
		2,4-Dichloromuconate	0.06	26 ± 3	3.3 ± 0.1	0.13

^aThe data for strains JMP 134, B13, and PRS2000 were calculated from data in references 52 and 53.

^bThe data for strains JMP 134, B13, and PRS2000 are from references 52 and 53.

^cND, not determined.

^dCMCI, chloromuconate cycloisomerase.