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Archives of Microbiology

, Volume 108, Issue 2, pp 195–202 | Cite as

Ribulose bisphosphate carboxylase from Thiobacillus A2. Its purification and properties

  • A. M. Charles
  • B. White
Article

Abstract

Ribulose bisphosphate carboxylase (EC 4.1.1.39) from Thiobacillus A2 has been purified to homogeneity on the basis of polyacrylamide gel electrophoresis and U.V. analysis during sedimentation velocity studies. The enzyme had an optimum pH of about 8.2 with Tris-HCl buffers. The molecular weight was about 521000 with an Srel. of 16.9. Km for RuBP was 122 μM, for total “CO2” it was 4.17 mM, and for Mg2+ 20.0 μM. The absolute requirement for a divalent cation was satisfied by Mg2+ which was replaceable to a certain extent by Mn2+. Activity was not significantly affected by SO 4 2- , SO 3 2- , or S2O 3 2- at 1.0 mM. At this concentration S2- caused a 27% stimulation. All mercurials tested were inhibitory. pHMB was the most potent causing about 60% inhibition at 0.01 mM. This inhibition was reversible by low concentrations of cysteine. Cyanide was also inhibitory. Its mode of inhibition with respect to RuBP was un-competitive and with a Ki of 20 μM. Lost activity could be restored partially by GSH or Cu2+. Although azide at the concentration tested had no significant effect on enzyme activity, 2,4-dinitrophenol at 1.0 mM caused 91% inhibition. Finally, activity was also affected by energy charge.

Key words

Enzymology CO2 fixation Thiobacilli Ribulose bisphosphate carboxylase Regulation Chemolithotrophy 

Abbreviations

ATP

adenosine-5′-triphosphate

GAPDH

glyceraldehyde phosphate dehydrogenase

GSH

(reduced) glutathione

G6P

glucose-6-phosphate

NAD+

nicotinamide adenine dinucleotide

NADP+

nicotinamide adenine dinucleotide phosphate

pHMB

parahydroxymercuribenzoate

6PG

6-phosphogluconate

3-PGA

3-phosphoglycerate

PGK

phosphoglyceratekinase

RuBP

ribulose-1,5-bisphosphate

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Copyright information

© Springer-Verlag 1976

Authors and Affiliations

  • A. M. Charles
    • 1
  • B. White
    • 1
  1. 1.Department of BiologyUniversity of WaterlooWaterlooCanada

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