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. 1970 Oct;104(1):211–218. doi: 10.1128/jb.104.1.211-218.1970

Characterization and Properties of the Pyruvate Phosphorylation System of Acetobacter xylinum

Moshe Benziman 1, A Palgi 1
PMCID: PMC248202  PMID: 4319721

Abstract

The enzyme responsible for the direct phosphorylation of pyruvate during gluconeogenesis in Acetobacter xylinum has been purified 46-fold from ultrasonic extracts and freed from interfering enzyme activities. The enzyme was shown to catalyze the reversible Mg2+ ion-dependent conversion of equimolar amounts of pyruvate, adenosine triphosphate (ATP), and orthophosphate (Pi) into phosphoenolpyruvate (PEP), adenosine monophosphate (AMP), and pyrophosphate (PP). The optimal pH for PEP synthesis was pH 8.2; for the reversal it was pH 6.5. The ratio between the initial rates of the reaction in the forward and reverse directions was 5.1 at pH 8.2 and 0.45 at pH 6.5. The apparent Km values of the components of the system in the forward reaction were: pyruvate, 0.2 mm; ATP, 0.4 mm; Pi, 0.8 mm; Mg2+, 2.2 mm; and for the reverse reaction: PEP, 0.1 mm; AMP, 1.6 μm; PP, 0.067 mm; Mg2+, 0.87 mm. PEP formation was inhibited by AMP and PP. The inhibition by AMP was competitive with regard to ATP (Ki = 0.2 mm). The reverse reaction was inhibited competitively by ATP and noncompetitively by pyruvate. The enzyme was strongly inhibited by p-hydroxymercuribenzoate. The inhibition was reversed by dithiothreitol and glutathione. The properties of the enzyme are discussed in relation to the regulation of the opposing enzymatic activities involved in the interconversion of PEP and pyruvate in A. xylinum.

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Selected References

These references are in PubMed. This may not be the complete list of references from this article.

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