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d-Ribulose-1,5-biphosphate carboxylase fromThiobacillus neapolitanus

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Abstract

d-Ribulose-1,5-bisphosphate carboxylase fromThiobacillus neapolitanus was isolated by differential centrifugation, sucrose density gradient centrifugation, and DEAE-Sephadex column chromatography. The specific activity of the purified enzyme was 2.8 μmol CO2 fixed/min/mg protein. The enzyme's homogeneity was indicated by a single migrating band during polyacrylamide disc gel electrophoresis and as a single symmetrical schlieren peak that sedimented at a constant rate during ultracentrifugation. TheS 20,w was 18.2; the molecular weight, 500,000±20.000. Sodium dodecyl sulfate polyacrylamide disc gel electrophoresis resolved two polypeptide chains of 55,000 and 11,000 daltons. The pH optimum 0f 7.75 with 9 mM MgCl2 shifted to 7.45 with 59 mM MgCl2. Enzyme dialyzed free of Mg++ was inactive and no other divalent cation substituted for Mg++. TheK m (Mg++),K m (CO2), andK m (RuBP) were 0.59 mM, 0.85 mM, and 0.092 mM, respectively. The inhibition by 6-phosphogluconate was competitive and no stimulation of activity could be demonstrated.

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  1. Department of Biochemistry, Clemson University, 29631, Clemson, South Carolina, USA

    R. M. Snead & J. M. Shively

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  1. R. M. Snead
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  2. J. M. Shively
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Snead, R.M., Shively, J.M. d-Ribulose-1,5-biphosphate carboxylase fromThiobacillus neapolitanus . Current Microbiology 1, 309–314 (1978). https://doi.org/10.1007/BF02601689

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