Propionyl-CoA carboxylase from Streptomyces coelicolor A3(2): cloning of the gene encoding the biotin-containing subunit.
In Streptomyces coelicolor A3(2), polyketides are made from malonyl-CoA, which is presumed to be derived from acetyl-CoA by the action of acetyl-CoA carboxylase (ACC). No ACC activity was found in cell-free extracts of S. coelicolor. However, propionyl-CoA carboxylase (PCC) activity was detected at substantial levels. Fixation of CO2 by ACC and PCC occurs by covalent bonding of CO2 to a biotin-containing protein. Most bacteria have a single small biotinylated protein of approximately 22 kDa, but S. coelicolor contains three larger biotin-containing proteins (approximately 145, 88 and 70 kDa). To determine which biotinylated protein was associated with PCC activity, the enzyme was purified and shown to comprise an alpha subunit (biotin-containing) of 88 kDa and a beta subunit of 66 kDa. The N-terminal sequences of these proteins were determined and, using an oligonucleotide probe, the gene for the alpha subunit (pccA) was cloned.
Bramwell
Helena
H
Divisions of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.|Divisions of Biochemistry & Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Hunter
Lain S
LS
Divisions of Molecular Genetics, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Coggins
John R
JR
Divisions of Biochemistry & Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
Nimmo
Hugh G
HG
Divisions of Biochemistry & Molecular Biology, Institute of Biomedical and Life Sciences, University of Glasgow, Glasgow G12 8QQ, UK.
D000595:Amino Acid Sequence; D001483:Base Sequence; D001710:Biotin; D002262:Carboxy-Lyases / Q000235:genetics*; D003001:Cloning, Molecular; D043586:Methylmalonyl-CoA Decarboxylase; D008969:Molecular Sequence Data; D010446:Peptide Fragments / Q000235:genetics; D015183:Restriction Mapping; D013302:Streptomyces / Q000201:enzymology* / Q000235:genetics
D010446:Peptide Fragments; D001710:Biotin; D002262:Carboxy-Lyases; D043586:Methylmalonyl-CoA Decarboxylase