Functional analysis of the gene cluster involved in production of the bacteriocin circularin A by Clostridium beijerinckii ATCC 25752.
A region of 12 kb flanking the structural gene of the cyclic antibacterial peptide circularin A of Clostridium beijerinckii ATCC 25752 was sequenced, and the putative proteins involved in the production and secretion of circularin A were identified. The genes are tightly organized in overlapping open reading frames. Heterologous expression of circularin A in Enterococcus faecalis was achieved, and five genes were identified as minimally required for bacteriocin production and secretion. Two of the putative proteins, CirB and CirC, are predicted to contain membrane-spanning domains, while CirD contains a highly conserved ATP-binding domain. Together with CirB and CirC, this ATP-binding protein is involved in the production of circularin A. The fifth gene, cirE, confers immunity towards circularin A when expressed in either Lactococcus lactis or E. faecalis and is needed in order to allow the bacteria to produce bacteriocin. Additional resistance against circularin A is conferred by the activity of the putative transporter consisting of CirB and CirD.
Kemperman
Robèr
R
Department of Molecular Genetics, Biomolecular Sciences and Biotechnology Institute, University of Groningen, Groningen, The Netherlands.
Jonker
Marnix
M
Nauta
Arjen
A
Kuipers
Oscar P
OP
Kok
Jan
J
D000595:Amino Acid Sequence; D001426:Bacterial Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics; D001430:Bacteriocins / Q000096:biosynthesis* / Q000235:genetics / Q000494:pharmacology; D001483:Base Sequence; D003013:Clostridium / Q000187:drug effects / Q000235:genetics / Q000254:growth & development / Q000378:metabolism*; D024881:Drug Resistance, Bacterial / Q000235:genetics; D013293:Enterococcus faecalis / Q000235:genetics / Q000378:metabolism; D013294:Lactococcus lactis / Q000235:genetics / Q000378:metabolism; D008826:Microbial Sensitivity Tests; D008969:Molecular Sequence Data; D005810:Multigene Family; D016366:Open Reading Frames; D017422:Sequence Analysis, DNA
D001426:Bacterial Proteins; D001430:Bacteriocins