Study of the (S)-hydroxynitrile lyase from Hevea brasiliensis:: mechanistic implications

被引：33

作者：

Hanefeld, U ^{[1
]}

Straathof, AJJ ^{[1
]}

Heijnen, JJ ^{[1
]}

机构：

[1] Delft Univ Technol, Kluyver Lab Biotechnol, NL-2628 BC Delft, Netherlands

来源：

BIOCHIMICA ET BIOPHYSICA ACTA-PROTEIN STRUCTURE AND MOLECULAR ENZYMOLOGY | 1999年 / 1432卷 / 02期

关键词：

hydroxynitrile lyase; oxynitrilase; electrospray mass spectrometry; catalytic triad; trans-cyanohydrin reaction; transcyanation; (S)-cyanohydrin; H-1-nuclear magnetic resonance; (Hevea brasiliensis);

D O I：

10.1016/S0167-4838(99)00108-9

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Investigations of the (S)-selective hydroxynitrile lyase from Hevea brasiliensis were performed by electrospray mass spectroscopy, H-1-NMR and with an enzyme activity assay. For the trans-cyanohydrin reaction (transcyanation) a two step reaction could be established. The results furthermore indicate a fast deactivation of the enzyme at low pH and a strong substrate dependence of its stability. They rule out an enzyme-HCN complex or a covalently bound carbonyl compound. Therefore the earlier postulated reaction intermediate as well as the proposed action of the catalytic triad have to be reevaluated. The calculated molecular mass could be confirmed by mass spectroscopy. (C) 1999 Elsevier Science B.V. All rights reserved.

引用

页码：185 / 193

页数：9

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