A thiocyanate hydrolase of Thiobacillus thioparus. A novel enzyme catalyzing the formation of carbonyl sulfide from thiocyanate
- Faculty of Agriculture, Tokyo University of Agriculture and Technology, (Japan)
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.
- OSTI ID:
- 5227051
- Journal Information:
- Journal of Biological Chemistry; (United States), Vol. 267:13; ISSN 0021-9258
- Country of Publication:
- United States
- Language:
- English
Similar Records
Characterization of the pyrogallol-phloroglucinol isomerase of Eubacterium oxidoreducens
87-kDa protein, a major specific substrate for protein kinase C: purification from bovine brain and characterization
Related Subjects
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