Activation studies of the β-carbonic anhydrases from Malassezia restricta with amines and amino acids

Figures & data

Figure 1. Amino acids and amines 1–24 investigated as CAAs in the present article.

Table 1. Activation of hCA I, II and MreCA with L-Trp, at 25 °C, for the CO₂ hydration reaction²⁵.

Enzyme	kcat^a	KM^a	(kcat)L-Trp^b	KA^c(μM)
	(s ^− 1)	(mM)	(s^−1)	L-Trp
hCA I^d	2.0 × 105	4.0	3.4 × 105	44.0
hCA II^d	1.4 × 106	9.3	4.9 × 106	27.0
MreCA^e	1.06 × 106	9.9	9.6 × 106	0.32

^aObserved catalytic rate without activator. K_M values in the presence and the absence of activators were the same for the various CAs (data not shown).

^bObserved catalytic rate in the presence of 10 μM activator.

^cThe activation constant (K_A) for each enzyme was obtained by fitting the observed catalytic enhancements as a function of the activator concentration²⁵. Mean from at least three determinations by a stopped-flow, CO₂ hydrase method²⁵. Standard errors were in the range of 5–10% of the reported values (data not shown).

^dHuman recombinant isozymes, from work by Capasso and Supuran³².

^eFungal recombinant enzyme, this work.

Table 2. Activation constants of hCA I, hCA II and MreCA with amino acids and amines 1–24 by a stopped-flow CO₂ hydrase assay²⁵.

No.	Compound	KA (μM)^a
		hCA I^b	hCA II^c	MreCA^c
1	L-His	0.03	10.9	12.8
2	D-His	0.09	43	1.84
3	L-Phe	0.07	0.013	2.69
4	D-Phe	86	0.035	0.76
5	L-DOPA	3.1	11.4	0.87
6	D-DOPA	4.9	7.8	0.70
7	L-Trp	44	27	0.32
8	D-Trp	41	12	0.89
9	L-Tyr	0.02	0.011	4.15
10	D-Tyr	0.04	0.013	7.83
11	4-H2N-L-Phe	0.24	0.15	0.61
12	Histamine	2.1	125	0.90
13	Dopamine	13.5	9.2	2.71
14	Serotonin	45	50	0.82
15	2-Pyridyl-methylamine	26	34	0.34
16	2–(2-Aminoethyl)pyridine	13	15	2.13
17	1–(2-Aminoethyl)-piperazine	7.4	2.3	0.25
18	4–(2-Aminoethyl)-morpholine	0.14	0.19	0.33
19	L-Adrenaline	0.09	96.0	0.015
20	L-Asn	11.3	>100	0.93
21	L-Asp	5.20	>100	4.04
22	L-Glu	6.43	>100	5.26
23	D-Glu	10.7	>100	4.70
24	L-Gln	>100	>50	0.90

^aMean from three determinations by a stopped-flow, CO₂ hydrase method²⁵. Standard errors were in the range of 5–10% of the reported values (data not shown).

^bHuman recombinant isozymes, from the work by Supran and collegues³.

^cFungal recombinant enzyme, this work.

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