1887

Abstract

Pyruvate decarboxylase (PDC), an enzyme central to homoethanol fermentation, catalyses the non-oxidative decarboxylation of pyruvate to acetaldehyde with release of carbon dioxide. PDC enzymes from diverse organisms have different kinetic properties, thermal stability and codon usage that are likely to offer unique advantages for the development of desirable Gram-positive biocatalysts for use in the ethanol industry. To examine this further, genes from bacteria to yeast were expressed in the Gram-positive host . The PDC activity and protein levels were determined for each strain. In addition, the levels of -specific mRNA transcripts and stability of recombinant proteins were assessed. From this analysis, the gene of Gram-positive was found to be the most advantageous for engineering high-level synthesis of PDC in a Gram-positive host. This gene was thus selected for transcriptional coupling to the alcohol dehydrogenase gene () of . The resulting Gram-positive ethanol production operon was expressed at high levels in . Extracts from this recombinant were shown to catalyse the production of ethanol from pyruvate.

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2005-12-01
2019-10-19
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