Cloning of genes coding for the three subunits of thiocyanate hydrolase of Thiobacillus thioparus THI 115 and their evolutionary relationships to nitrile hydratase

Yoko Katayama, Yasuhiko Matsushita, Miyuki Kaneko, Mai Kondo, Tadayoshi Mizuno, Hiroshi Nyunoya

Research output: Contribution to journalArticle

62 Citations (Scopus)

Abstract

Thiocyanate hydrolase is a newly found enzyme from Thiobacillus thioparus THI 115 that converts thiocyanate to carbonyl sulfide and ammonia (Y. Katayama, Y. Narahara, Y. Inoue, F. Amano, T. Kanagawa, and H. Kuraishi, J. Biol. Chem. 267:9170-9175, 1992). We have cloned and sequenced the scn genes that encode the three subunits of the enzyme. The scnB, scnA, and scnC genes, arrayed in this order, contained open reading frames encoding sequences of 157, 126, and 243 amino acid residues, respectively, for the β, α, and γ subunits, respectively. Each open reading frame was preceded by a typical Shine-Dalgarno sequence. The deduced amino-terminal peptide sequences for the three subunits were in fair agreement with the chemically determined sequences. The protein molecular mass calculated for each subunit was compatible with that determined by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. From a computer analysis, thiocyanate hydrolase showed significant homologies to bacterial nitrile hydratases known to convert nitrile to the corresponding amide, which is further hydrolyzed by amidase to form acid and ammonia. The two enzymes were homologous over regions corresponding to almost the entire coding regions of the genes: the β and α subunits of thiocyanate hydrolase were homologous to the amino- and carboxyl- terminal halves of the β subunit of nitrile hydratase, and the γ subunit of thiocyanate hydrolase was homologous to the α subunit of nitrile hydratase. Comparisons of the catalytic properties of the two homologous enzymes support the model for the reaction steps of thiocyanate hydrolase that was previously presented on the basis of biochemical analyses.

Original languageEnglish
Pages (from-to)2583-2589
Number of pages7
JournalJournal of Bacteriology
Volume180
Issue number10
Publication statusPublished - 1998 May
Externally publishedYes

Fingerprint

Thiobacillus
Organism Cloning
Genes
Enzymes
amidase
Ammonia
Open Reading Frames
Nitriles
Amides
Sodium Dodecyl Sulfate
Polyacrylamide Gel Electrophoresis
nitrile hydratase
thiocyanate hydrolase
Amino Acids
Peptides
Acids
Proteins

ASJC Scopus subject areas

  • Applied Microbiology and Biotechnology
  • Immunology

Cite this

Cloning of genes coding for the three subunits of thiocyanate hydrolase of Thiobacillus thioparus THI 115 and their evolutionary relationships to nitrile hydratase. / Katayama, Yoko; Matsushita, Yasuhiko; Kaneko, Miyuki; Kondo, Mai; Mizuno, Tadayoshi; Nyunoya, Hiroshi.

In: Journal of Bacteriology, Vol. 180, No. 10, 05.1998, p. 2583-2589.

Research output: Contribution to journalArticle

Katayama, Yoko ; Matsushita, Yasuhiko ; Kaneko, Miyuki ; Kondo, Mai ; Mizuno, Tadayoshi ; Nyunoya, Hiroshi. / Cloning of genes coding for the three subunits of thiocyanate hydrolase of Thiobacillus thioparus THI 115 and their evolutionary relationships to nitrile hydratase. In: Journal of Bacteriology. 1998 ; Vol. 180, No. 10. pp. 2583-2589.
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