1887

Abstract

grows in a remarkable range of mesophilic environments from pH 2 to pH 10. During growth in acidic environments, where acetate is toxic, expression of pyruvate decarboxylase (PDC) serves to direct the flow of pyruvate into ethanol during fermentation. PDC is rare in bacteria and absent in animals, although it is widely distributed in the plant kingdom. The gene from is the first to be cloned and characterized from a Gram-positive bacterium. In , the recombinant gene from was poorly expressed due to differences in codon usage that are typical of low-G+C organisms. Expression was improved by the addition of supplemental codon genes and this facilitated the 136-fold purification of the recombinant enzyme as a homo-tetramer of 58 kDa subunits. Unlike PDC, which exhibits Michaelis–Menten kinetics, PDC is activated by pyruvate and exhibits sigmoidal kinetics similar to fungal and higher plant PDCs. Amino acid residues involved in the allosteric site for pyruvate in fungal PDCs were conserved in PDC, consistent with a conservation of mechanism. Cluster analysis of deduced amino acid sequences confirmed that PDC is quite distant from PDC and plant PDCs. PDC appears to have diverged very early from a common ancestor which included most fungal PDCs and eubacterial indole-3-pyruvate decarboxylases. These results suggest that the gene is quite ancient in origin, in contrast to the , which may have originated by horizontal transfer from higher plants.

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2001-09-01
2019-10-23
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