Synthesis of amidrazones using an engineered papain nitrile hydratase

FEBS Letters

Nägler, Dorit K¹  Dufour, Éric¹  Ménard, Robert¹  Storer, Andrew C¹  Tam, Wendy¹ 

[1] Biotechnology Research Institute, National Research Council of Canada, 6100 Avenue Royalmount, Montreal, Que. H4P 2R2, Canada

关键词: Papain;    Cysteine protease;    Nitrile hydratase;    Nitrilase;    Biocatalysis;    Protein engineering;   

DOI  :  10.1016/S0014-5793(98)00773-X

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

To demonstrate the usefulness of an engineered papain nitrile hydratase as a biocatalyst, a peptide amidrazone was prepared by incubation of the nitrile MeOCO-Phe-Ala-nitrile with the Gln¹⁹Glu 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.

【 授权许可】

Unknown