Archives of Microbiology

, Volume 146, Issue 2, pp 105–110

Pyruvate decarboxylase from Zymomonas mobilis. Isolation and partial characterization

  • S. Bringer-Meyer
  • K. -L. Schimz
  • H. Sahm
Original Papers

DOI: 10.1007/BF00402334

Cite this article as:
Bringer-Meyer, S., Schimz, K.L. & Sahm, H. Arch. Microbiol. (1986) 146: 105. doi:10.1007/BF00402334

Abstract

Pyruvate decarboxylase (EC 4.1.1.1) from the ethanol producing bacterium Zymomonas mobilis was purified to homogeneity. This enzyme is an acidic protein with an isoelectric point of 4.87 and has an apparent molecular weight of 200,000±10,000. The enzyme showed a single band in sodium dodecylsulfate gel electrophoresis with a molecular weight of 56,500±4,000 which indicated that the enzyme consists of four probably identical subunits. The dissociation of the cofactors Mg2+ and thiamine pyrophosphate at pH 8.9 resulted in a total loss of enzyme activity which could be restored to 99.5% at pH 6.0 in the presence of both cofactors. For the apoenzyme the apparent Km values for Mg2+ and thiamine pyrophosphate were determined to be 24 μM and 1.28 μM. The apparent Km value for the substrate pyruvate was 0.4 mM. Antiserum prepared against this purified pyruvate decarboxylase failed to crossreact with cell extracts of the reportedly pyruvate decarboxylase positive bacteria Sarcina ventriculi, Erwinia amylovora, or Gluconobacter oxydans, or with cell extracts of Saccharomyces cerevisiae.

Key words

Pyruvate decarboxylase Zymomonas mobilis Purification Molecular weight Isoelectric point Cofactor dissociation Immunological studies 

Abbreviations

Tris-buffer

0,01 M tris-HCl buffer, containing 1 mM MgCl2 0.1 mM EDTA, 1.0 mM thiamine pyrophosphate, 2 mM mercaptopropanediol, pH 7.0

Copyright information

© Springer-Verlag 1986

Authors and Affiliations

  • S. Bringer-Meyer
    • 1
  • K. -L. Schimz
    • 1
  • H. Sahm
    • 1
  1. 1.Institut für Biotechnologie 1 der Kernforschungsanlage Jülich GmbHJülichGermany