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Title: A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate

Abstract

A thiocyanate hydrolase that catalyzes the first step in thiocyanate degradation was purified to homogeneity from Thiobacillus thioparus, an obligate chemolithotrophic eubacterium metabolizing thiocyanate to sulfate as an energy source. The thiocyanate hydrolase was purified 52-fold by steps involving ammonium sulfate precipitation, DEAE-Sephacel column chromatography, and hydroxylapatite column chromatography. The enzyme hydrolyzed 1 mol of thiocyanate to form 1 mol of carbonyl sulfide and 1 mol of ammonia as follows: SCN- + 2H2O----COS + NH3 + OH-. This is the first report describing the hydrolysis of thiocyanate to carbonyl sulfide by an enzyme. The enzyme had a molecular mass of 126 kDa and was composed of three different subunits: alpha (19 kDa), beta (23 kDa), and gamma (32 kDa). The enzyme exhibited optimal activities at pH 7.5-8.0 and at temperatures ranging from 30 to 40 degrees C. The Km value for thiocyanate was approximately 11 mM. Immunoblot analysis with polyclonal antibodies against the purified enzyme suggested that it was induced in T. thioparus cells when the cells were grown with thiocyanate.

Authors:
; ; ; ; ;  [1]
  1. Faculty of Agriculture, Tokyo University of Agriculture and Technology, (Japan)
Publication Date:
OSTI Identifier:
5227051
Resource Type:
Journal Article
Journal Name:
Journal of Biological Chemistry; (United States)
Additional Journal Information:
Journal Volume: 267:13; Journal ID: ISSN 0021-9258
Country of Publication:
United States
Language:
English
Subject:
63 RADIATION, THERMAL, AND OTHER ENVIRON. POLLUTANT EFFECTS ON LIVING ORGS. AND BIOL. MAT.; HYDROLASES; ENZYME ACTIVITY; SULFIDES; BIOSYNTHESIS; THIOCYANATES; METABOLISM; BACTERIA; CATALYSIS; ELECTROPHORESIS; LIQUID COLUMN CHROMATOGRAPHY; MOLECULAR WEIGHT; ANTITHYROID DRUGS; CARBONIC ACID DERIVATIVES; CHALCOGENIDES; CHROMATOGRAPHY; DRUGS; ENZYMES; HORMONE ANTAGONISTS; MICROORGANISMS; ORGANIC COMPOUNDS; ORGANIC SULFUR COMPOUNDS; PROTEINS; SEPARATION PROCESSES; SULFUR COMPOUNDS; SYNTHESIS; 560300* - Chemicals Metabolism & Toxicology

Citation Formats

Katayama, Y, Narahara, Y, Inoue, Y, Amano, F, Kanagawa, T, and Kuraishi, H. A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate. United States: N. p., 1992. Web.
Katayama, Y, Narahara, Y, Inoue, Y, Amano, F, Kanagawa, T, & Kuraishi, H. A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate. United States.
Katayama, Y, Narahara, Y, Inoue, Y, Amano, F, Kanagawa, T, and Kuraishi, H. 1992. "A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate". United States.
@article{osti_5227051,
title = {A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate},
author = {Katayama, Y and Narahara, Y and Inoue, Y and Amano, F and Kanagawa, T and Kuraishi, H},
abstractNote = {A thiocyanate hydrolase that catalyzes the first step in thiocyanate degradation was purified to homogeneity from Thiobacillus thioparus, an obligate chemolithotrophic eubacterium metabolizing thiocyanate to sulfate as an energy source. The thiocyanate hydrolase was purified 52-fold by steps involving ammonium sulfate precipitation, DEAE-Sephacel column chromatography, and hydroxylapatite column chromatography. The enzyme hydrolyzed 1 mol of thiocyanate to form 1 mol of carbonyl sulfide and 1 mol of ammonia as follows: SCN- + 2H2O----COS + NH3 + OH-. This is the first report describing the hydrolysis of thiocyanate to carbonyl sulfide by an enzyme. The enzyme had a molecular mass of 126 kDa and was composed of three different subunits: alpha (19 kDa), beta (23 kDa), and gamma (32 kDa). The enzyme exhibited optimal activities at pH 7.5-8.0 and at temperatures ranging from 30 to 40 degrees C. The Km value for thiocyanate was approximately 11 mM. Immunoblot analysis with polyclonal antibodies against the purified enzyme suggested that it was induced in T. thioparus cells when the cells were grown with thiocyanate.},
doi = {},
url = {https://www.osti.gov/biblio/5227051}, journal = {Journal of Biological Chemistry; (United States)},
issn = {0021-9258},
number = ,
volume = 267:13,
place = {United States},
year = {Tue May 05 00:00:00 EDT 1992},
month = {Tue May 05 00:00:00 EDT 1992}
}