Expression of the cyanide hydratase enzyme from Fusarium lateritium in Escherichia coli and identification of an essential cysteine residue.
The filamentous fungus Fusarium lateritium is cyanide tolerant, due partly to the induction of the enzyme cyanide hydratase in the presence of cyanide. This enzyme catalyses the hydration of cyanide to formamide. The expression in Escherichia coli of a cDNA clone encoding cyanide hydratase is described. The cDNA cloned was expressed as a transcriptional fusion in the expression vector pKK233-2 and a high level of activity of cyanide hydratase was detected in E. coli. Site-directed mutagenesis of the cys-163 residue inactivated the enzyme.
Brown
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School of Health Sciences, University of Sunderland, UK.
Turner
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O'Reilly
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D000595:Amino Acid Sequence; D001483:Base Sequence; D001665:Binding Sites; D003001:Cloning, Molecular; D003486:Cyanides / Q000378:metabolism; D003545:Cysteine / Q000737:chemistry; D018076:DNA, Complementary / Q000235:genetics; D004271:DNA, Fungal / Q000235:genetics; D004926:Escherichia coli / Q000201:enzymology / Q000235:genetics; D005670:Fusarium / Q000201:enzymology* / Q000235:genetics*; D005800:Genes, Fungal; D006836:Hydro-Lyases / Q000737:chemistry / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D016297:Mutagenesis, Site-Directed; D011994:Recombinant Proteins / Q000737:chemistry / Q000235:genetics / Q000378:metabolism
D003486:Cyanides; D018076:DNA, Complementary; D004271:DNA, Fungal; D011994:Recombinant Proteins; D006836:Hydro-Lyases; C076848:cyanide hydratase; D003545:Cysteine