FEBS Letters | |
From β‐glucanase to β‐glucansynthase: glycosyl transfer to α‐glycosyl fluorides catalyzed by a mutant endoglucanase lacking its catalytic nucleophile | |
Planas, Antoni1  Malet, Carles1  | |
[1] Laboratory of Biochemistry, Institut Quı́mic de Sarrià, Universitat Ramon Llull, Via Augusta 390, 08017 Barcelona, Spain | |
关键词: Glycosynthase; Enzymatic glycosylation; β-Glucanase; Glycosyl fluoride; Nucleophile residue; 1; 3-1; 4-β-glucanase; 1; 3-1; 4-β-d-glucan 4-glucanohydrolase; MU; 4-methylumbelliferyl; MALDI-TOF; matrix-assisted laser desorption ionization time of flight; MES; 2-[N-morpholino]ethanesulfonic acid; TES; N-tris[hydroxymethyl]methyl-2-aminoethanesulfonic acid; | |
DOI : 10.1016/S0014-5793(98)01448-3 | |
学科分类:生物化学/生物物理 | |
来源: John Wiley & Sons Ltd. | |
【 摘 要 】
Removal of the catalytic nucleophile Glu134 of the retaining 1,3-1,4-β-glucanase from Bacillus licheniformis by mutation to alanine yields an enzyme with no glycosidase activity. The mutant is able to catalyze the regio- and stereospecific glycosylation of α-laminaribiosyl fluoride with different glucoside acceptors through a single-step inverting mechanism. The main advantage of the mutant as glycosylation catalyst with respect to the kinetically controlled transglycosylation using the wild-type enzyme is that the reaction products cannot be hydrolyzed by the mutant enzyme, and glycosylation yields rise to 90%.
【 授权许可】
Unknown
【 预 览 】
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RO201912020306912ZK.pdf | 198KB | download |