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Phosphoenolpyruvate carboxylase from Acetobacter aceti

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Abstract

In Acetobacter aceti growing on pyruvate as the only source of carbon and energy, oxaloacetate (OAA) is produced by a phosphoenolpyruvate (PEP) carboxylase (EC 4.1.1.31). The enzyme was purified 122-fold and a molecular weight of about 380,000 was estimated by gel filtration.

The optimum pH was 7.5 and the K m values for PEP and NaHCO3 were 0.49 mM and about 3 mM, respectively. The enzyme needed a divalent cation; the K m for Mn2+, Co2+ and Mg2+ were 0.12, 0.26 and 0.77 mM, respectively. Maximal activity was only obtained with Mg2+. Mn2+ and Co2+ became inhibitory at high concentrations.

The activity was inhibited by succinate and, to a lesser extent, by fumarate, citrate, α-ketoglutarate, aspartate and glutamate.

As compared with the corresponding enzyme from A. xylinum, the PEP carboxylase of A. aceti showed the following differences: a) It had an absolute requirement for acetyl CoA (K a 0.18 mM) or propionyl CoA (K a 0.2 mM). b) It was not affected by ADP. c) It was sensitive to thiol blocking agents.

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Abbreviations

PEP:

phosphoenolpyruvate

OAA:

oxaloacetate

MW:

molecular weight

TEMG buffer:

50 mM Tris-HCl, pH 7.5, 1 mM EDTA, 5 mM MgCl2, 1 mM glutathione

HEPES:

N-2-hydroxyethylpiperazine-N′-ethanesulfonic acid

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Author notes

  1. J. -P. Schwitzguébel

    Present address: Department of Botany, Imperial College, SW7 2BB, London, England

Authors and Affiliations

  1. Eidgenössische Technische Hochschule, ETH-Zentrum, Mikrobiologisches Institut, CH-8092, Zürich, Switzerland

    J. -P. Schwitzguébel & L. Ettlinger

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  1. J. -P. Schwitzguébel
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  2. L. Ettlinger
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Schwitzguébel, J.P., Ettlinger, L. Phosphoenolpyruvate carboxylase from Acetobacter aceti . Arch. Microbiol. 122, 109–115 (1979). https://doi.org/10.1007/BF00408053

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  • DOI: https://doi.org/10.1007/BF00408053

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