Kinetic and paramagnetic NMR investigations of the inhibition of Streptomyces antibioticus tyrosinase
Kinetic and paramagnetic NMR investigations of the inhibition of Streptomyces antibioticus tyrosinase
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DOI：
10.1016/s1381-1177(99)00064-8
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发表时间：
2000-01-12
影响因子：
--
通讯作者：
Canters, GW
中科院分区：
文献类型：
--
作者：
Bubacco, L;Vijgenboom, E;Canters, GW
A scaled-up isolation and purification procedure is described for tyrosinase from Streptomyces antibioticus. Kinetic studies of the enzyme catalysed conversion of L-3,4-dihydroxyphenylalanine (L-DOPA) into DOPAchrome show that kojic acid, L-mimosine, p-toluic acid and benzoic acid exhibit competitive inhibition with inhibition constants of 3.4, 30, 1.9 X 10(2) and 8.0 X 10(2) mu M, respectively. Paramagnetic NMR techniques appear well suited to study the binding of inhibitors to the active site. From the variation of the NMR shifts with temperature a value of -2J = 156 +/- 6 cm(-1) is derived for the exchange coupling between the unpaired spins on the two Cu(II) ions in the active site of the met-tyrosinase/kojic acid complex. (C) 2000 Elsevier Science B.V. All rights reserved.
