Synthesis of amidrazones using an engineered papain nitrile hydratase

被引:13
作者
Dufour, E [1 ]
Tam, W [1 ]
Nägler, DK [1 ]
Storer, AC [1 ]
Ménard, R [1 ]
机构
[1] Natl Res Council Canada, Biotechnol Res Inst, Montreal, PQ H4P 2R2, Canada
关键词
papain; cysteine protease; nitrile hydratase; nitrilase; biocatalysis; protein engineering;
D O I
10.1016/S0014-5793(98)00773-X
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
To demonstrate the usefulness of an engineered papain nitrile hydratase as a biocatalyst, a peptide amidrazone mas prepared by incubation of the nitrile MeOCO-Phe-Ala-nitrile with the Gln(19)Glu papain mutant in the presence of salicylic hydrazide as a nucleophile, The amidrazone results from nucleophilic attack by salicylic hydrazide at the imino carbon of the thioimidate adduct formed between the enzyme and the peptide nitrile substrate, Compared to wild-type enzyme, the engineered nitrile hydratase causes a better than 4000-fold increase in the rate of amidrazone formation and yields a product of much higher purity. The advantages over other nitrile-hydrolyzing enzymes and current limitations of the papain nitrile hydratase are discussed. Published by Elsevier on behalf of the Federation of European Biochemical Societies.
引用
收藏
页码:78 / 82
页数:5
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