Stereochemical course of hydrolysis catalyzed by arabinofuranosyl hydrolases

FEBS Letters

Voragen, Alphons G.J.¹  Pitson, Stuart M.¹  Beldman, Gerrit¹ 

[1] Department of Food Science, Wageningen Agricultural University, Bomenweg 2, 6703 HD Wageningen, The Netherlands

关键词: Endo-(1 → 5)-α-l-Arabinanase;    α-l-Arabinofuranosidase;    Arabinoxylan arabinohydrolase;    Hydrolysis mechanism;    Aspergillus niger;    Glycosyl hydrolase family;    AXH;    arabinoxylan arabinohydrolase;    pNPA;    4-nitrophenyl α-l-arabinofuranoside;    HPAEC;    high performance anion exchange chromatography;   

DOI  :  10.1016/S0014-5793(96)01153-2

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The stereochemical course of hydrolysis catalyzed by various enzymes acting on arabinofuranosyl linkages has been determined. ¹H-NMR analysis of the action of endo-(1 → 5)-α-l-arabinanases from Aspergillus niger and Aspergillus aculeatus showed that both hydrolyze linear arabinan with inversion of configuration, and may therefore act via a single displacement mechanism. This is consistent with the A. niger enzyme's classification in glycosyl hydrolase family 43. The catalytic mechanisms of α-l-arabinofuranosidases from A. niger, A. aculeatus, Aspergillus awamori, Humicola insolens, Penicillium capsulatum and Bacillus subtilis were investigated using both ¹H-NMR and high performance anion exchange chromatography to follow glycosyl transfer reactions to methanol. In all cases these enzymes catalyzed the reaction with retention of configuration, and therefore probably operate via double displacement hydrolytic mechanisms. From the results with arabinofuranosidase A and B from A. niger we predict that all members of glycosyl hydrolase family 51 and 54 catalyze hydrolysis with net retention of anomeric configuration. Similar studies with (1 → 4)-β-d-arabinoxylan arabinohydrolases from A. awamori, Trichoderma reesei and Bifidobacterium adolescentis only enabled their tentative classification as inverting enzymes on the basis of their lack of glycosyl transfer to methanol.

【 授权许可】

Unknown