Enzymatic hydrolysis of cyanohydrins with recombinant nitrile hydratase and amidase from Rhodococcus erythropolis

• Published in: Biotechnology letters Vol. 26; no. 21; pp. 1675 - 1680
• Main Authors: REISINGER, Ch, OSPRIAN, I, GLIEDER, A, SCHOEMAKER, H. E, GRIENGL, H, SCHWAB, H
• Format: Journal Article
• Language: English
• Published: Dordrecht Springer 01.11.2004
• Subjects: Amidohydrolases - chemistry; Amidohydrolases - genetics; Amidohydrolases - metabolism; Biological and medical sciences; Biotechnology; Cloning, Molecular - methods; Escherichia coli - enzymology; Escherichia coli - genetics; Fundamental and applied biological sciences. Psychology; Hydro-Lyases - chemistry; Hydro-Lyases - genetics; Hydro-Lyases - metabolism; Hydrolysis; Nitriles - chemistry; Nitriles - metabolism; Protein Engineering - methods; Recombinant Proteins - biosynthesis; Recombinant Proteins - chemistry; Rhodococcus - classification; Rhodococcus - enzymology; Rhodococcus - genetics; Rhodococcus erythropolis; Enzyme; Amidase; Rhodococcus erythropolis; Lyases; Hydrolysis; cyanohydrin hydrolysis; Cyanohydrin; Biotransformation; Nitrile hydratase; Bacteria; Hydrolases; Actinomycetes; Carbon-oxygen lyases; Hydro-lyases; Enzymatic reaction
• ISSN: 0141-5492; 1573-6776
• DOI: 10.1007/s10529-004-3521-4

Nitrile hydratase and amidase from Rhodococcus erythropolis CIMB11540 were both cloned and expressed in Escherichia coli. Crude cell free extracts were used for the hydrolysis of different aromatic cyanohydrins. Nitrile hydratase expression was increased up to 5-fold by redesign of the expression cassette. The recombinant enzymes were successfully used for the conversion of several cyanohydrins to the corresponding alpha-hydroxy amides and acids while retaining enantiopurity.