Studies on Thermophilic Sulfate-Reducing Bacteria Iii

Abstract

AKAGI, J. AM. (University of Kansas, Lawrence) AND L. LEON CAMPBELL. Studies on thermophilic sulfate-reducing bacteria. III. Adenosine triphosphate-sulfurylase of Clostridium nigrificans and Desulfovibrio desulfuricans. J. Bacteriol. 84:1194-1201. 1962.-Adenosine triphosphate (ATP)-sulfurylase, which catalyzes the formation of adenosine-5'-phosphosulfate (APS) from ATP and S04=, has been purified from crude extracts of Clostridium nigrificans and Desulfovibrio desulfuricans by (NH4) SSO4 fractionation and triethylaminoethyl column chromatography. The enzyme from both sources operates over a broad pH range from 6.0 to 9.5. Below pH 6.0, activity decreases sharply, with no detectable activity at pH 5.0. Of the nucleotides tested (ATP and the triphosphates of deoxyadenosine, uridine, inosine, and guanosine), only ATP was acted upon by the enzyme from either source. The enzyme requires Mg++ for activity. Incubation of the enzyme from both organisms with ATP and S3504in the presence of helium resulted in the formation of an S35labeled nucleotide whose electrophoretic mobility was identical to that of chemically prepared APS. When incubated with ATP and the group VI anions (CrO4, MoO4, W04), the enzyme from both organisms formed an unstable intermediate, resulting in the accumulation of pyrophosphate. Thermal stability studies revealed that the ATP-sulfurylase of C. nigrificans was stable at higher temperatures than the enzyme obtained I Presented at the 62nd Annual Meeting of the

Desulfovibrio africanus sp. n., a New Dissimilatory Sulfate-reducing Bacterium

“…The Walvis Bay strain was originally isolated from a marine mud sample from Walvis Bay, S. W. Africa. Both strains were obtained from the 1 Present address: University of Sussex, Falmer, Sussex, England.…”

Purification and characterization of ATP sulfurylase from the extremely thermophilic archaebacterial sulfate-reducer, Archaeoglobus fulgidus

“…ATP sulfurylase was active in Tris-malate and Tris-HC1 buffers between pH 5.5 and 9.0 with an optimum at pH 8.0. The broad pH range and the slightly alkaline pH optimum are common characteristics of the ATP sulfurylases so far examined [7,11,24,[27][28][29]31].…”

“…It has also been proven to play an important role in oxidative dissimilatory sulfur metabolism in thiobacilli [4][5][6] and some phototrophic sulfur bacteria [7,8]. Already in the early sixties ATP sulfurylases were shown and some of their properties examined in dissimilatory sulfate-reducing bacteria [9][10][11], however very little is known about their physical, chemical and kinetic characteristics. In this report we describe the purification of the first ATP sulfurylase of archacbacterial origin and examine a number of its properties.…”

“…In this report we describe the purification of the first ATP sulfurylase of archacbacterial origin and examine a number of its properties. We further present evidence for the existence of inorganic pyrophosphatase (EC 3.6.1.1) in A. fulgidus, the enzyme that is believed to pull the thermodynamically unfavourable ATP sulfurylase reaction (The equilibrium constant of the reaction amounts to about 10 -s [11][12][13]) in the direction of sulfate activation [141.…”

Separation and distribution of thiosulfate-oxidizing enzyme, tetrathionate reductase, and thiosulfate reductase in extracts of marine heterotroph strain 16B

“…Liquid cultures and 10 g of lyophilized cells were a generous gift of J. M. Akagi. The organism was maintained and grown in mass culture as described previously (1).…”