Genetic and biochemical characterization of the α and β components of a propionyl-CoA carboxylase complex of A3(2)

The GenBank accession numbers for the , and sequences determined in this work are AF113603, AF113604 and AF113605, respectively.

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Abstract

Two genes, and , with nearly identical nucleotide sequences were cloned from A3(2). The deduced amino acid sequences of the product of these two genes showed high similarity to BcpA2 of and other biotin-containing proteins from different organisms assumed to be the α subunit of a propionyl-CoA carboxylase. A gene, , encoding the carboxyl transferase subunit of this enzyme complex was also characterized. Strains disrupted in did not show any change in acetyl- or propionyl-CoA carboxylase activity, whilst cell-free extracts of a mutant strain contained a reduced level of propionyl-CoA carboxylase. No mutants in could be isolated, suggesting that the gene may be essential. Heterologous expression of , and in and reconstitution of enzyme activity confirmed that PccB is the β subunit of a propionyl-CoA carboxylase and that either AccA1 or AccA2 could act as the α component of this enzyme complex. The fact that mutants appear to be inviable suggests that this gene encodes a biotinylated protein that might be shared with other carboxyl transferases essential for the growth of .

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1999-11-01
2024-03-29
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