Isolation of assimilatroy- and dissimilatory-type sulfite reductases from Desulfovibrio vulgaris.
Bisulfite reductase (desulfoviridin) and an assimilatory sulfite reductase have been purified from extracts of Desulfovibrio vulgaris. The bisulfite reductase has absorption maxima at 628, 580, 408, 390, and 279 nm, and a molecular weight of 226,000 by sedimentation equilibrium, and was judged to be free of other proteins by disk electrophoresis and ultracentrifugation. On gels, purified bisulfite reductase exhibited two green bands which coincided with activity and protein. The enzyme appears to be a tetramer but was shown to have two different types of subunits having molecular weights of 42,000 and 50,000. The chromophore did not form an alkaline ferrohemochromogen, was not reduced with dithionite or borohydride, and did not form a spectrally visible complex with CO. The assimilatory sulfite reductase has absorption maxima at 590, 545, 405 and 275 nm and a molecular weight of 26,800, and appears to consist of a single polypeptide chain as it is not dissociated into subunits by sodium dodecyl sulfate. By disk electrophoresis, purified sulfite reductase exhibited a single greenish-brown band which coincided with activity and protein. The sole product of the reduction was sulfide, and the chromophore was reduced by borohydride in the presence of sulfite. Carbon monoxide reacted with the reduced chromophore but it did not form a typical pyridine ferrohemochromogen. Thiosulfate, trithionate, and tetrathionate were not reduced by either enzyme preparation. In the presence of 8 M urea, the spectrum of bisulfite reductase resembles that of the sulfite reductase, thus suggesting a chemical relationship between the two chromophores.
D001426:Bacterial Proteins / Q000032:analysis; D001894:Borohydrides / Q000494:pharmacology; D002248:Carbon Monoxide / Q000494:pharmacology; D002474:Cell-Free System; D011232:Chemical Precipitation; D002848:Chromatography, DEAE-Cellulose; D002850:Chromatography, Gel; D003901:Desulfovibrio / Q000201:enzymology*; D004589:Electrophoresis, Disc; D006859:Hydrogen / Q000378:metabolism; D008365:Manometry; D008970:Molecular Weight; D010084:Oxidation-Reduction; D010088:Oxidoreductases / Q000032:analysis / Q000302:isolation & purification* / Q000378:metabolism; D013053:Spectrophotometry; D013440:Sulfides / Q000096:biosynthesis; D013447:Sulfites / Q000378:metabolism; D013885:Thiosulfates / Q000378:metabolism; D014461:Ultracentrifugation; D014508:Urea / Q000494:pharmacology
D001426:Bacterial Proteins; D001894:Borohydrides; D013440:Sulfides; D013447:Sulfites; D013885:Thiosulfates; D002248:Carbon Monoxide; D006859:Hydrogen; D014508:Urea; D010088:Oxidoreductases