An acyl-CoA dehydrogenase is involved in the formation of the Delta cis3 double bond in the acyl residue of the lipopeptide antibiotic friulimicin in Actinoplanes friuliensis.
The lipopeptide antibiotic friulimicin, produced by Actinoplanes friuliensis, is an effective drug against Gram-positive bacteria, such as methicillin-resistant Staphylococcus epidermidis and Staphylococcus aureus strains. Friulimicin consists of a cyclic peptide core of ten amino acids and an acyl residue linked to an exocyclic amino acid. The acyl residue is essential for antibiotic activity, varies in length from C13 to C15, and carries a characteristic double bond at position Delta cis3. Sequencing of a DNA fragment adjacent to a previously described fragment encoding some of the friulimicin biosynthetic genes revealed several genes whose gene products resemble enzymes of lipid metabolism. One of these genes, lipB, encodes an acyl-CoA dehydrogenase homologue. To elucidate the function of the LipB protein, a lipB insertion mutant was generated and the friulimicin derivative (FR242) produced by the mutant was purified. FR242 had antibiotic activity lower than friulimicin in a bioassay. Gas chromatography showed that the acyl residue of wild-type friulimicin contains a double bond, whereas a saturated bond was present in FR242. These results were confirmed by the heterologous expression of lipB in Streptomyces lividans T7, which led to the production of unsaturated fatty acids not found in the S. lividans T7 parent strain. These results indicate that the acyl-CoA dehydrogenase LipB is involved in the introduction of the unusual Delta cis3 double bond into the acyl residue of friulimicin.
Heinzelmann
Eva
E
Fakultät Biologie, Mikrobiologisches Institut, Mikrobiologie/Biotechnologie, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.
Berger
Susanne
S
Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie-Hans-Knöll-Institut, Beutenbergstrasse 11, 07745 Jena, Germany.
Müller
Claudia
C
Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie-Hans-Knöll-Institut, Beutenbergstrasse 11, 07745 Jena, Germany.
Härtner
Thomas
T
Fakultät Biologie, Mikrobiologisches Institut, Mikrobiologie/Biotechnologie, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.
Poralla
Karl
K
Fakultät Biologie, Mikrobiologisches Institut, Mikrobiologie/Biotechnologie, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.
Wohlleben
Wolfgang
W
Fakultät Biologie, Mikrobiologisches Institut, Mikrobiologie/Biotechnologie, Eberhard-Karls-Universität Tübingen, Auf der Morgenstelle 28, 72076 Tübingen, Germany.
Schwartz
Dirk
D
Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie-Hans-Knöll-Institut, Beutenbergstrasse 11, 07745 Jena, Germany.
D042964:Acyl-CoA Dehydrogenase / Q000378:metabolism*; D000900:Anti-Bacterial Agents / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism / Q000494:pharmacology; D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins / Q000235:genetics*; D002849:Chromatography, Gas; D003001:Cloning, Molecular; D004269:DNA, Bacterial / Q000737:chemistry / Q000235:genetics; D024881:Drug Resistance, Bacterial; D005231:Fatty Acids, Unsaturated / Q000032:analysis / Q000302:isolation & purification; D017353:Gene Deletion; D023061:Gene Order; D005798:Genes, Bacterial; D008848:Micromonosporaceae / Q000201:enzymology*; D008969:Molecular Sequence Data; D015394:Molecular Structure; D016254:Mutagenesis, Insertional; D010455:Peptides / Q000737:chemistry / Q000302:isolation & purification / Q000378:metabolism* / Q000494:pharmacology; D017422:Sequence Analysis, DNA; D017386:Sequence Homology, Amino Acid; D048170:Streptomyces lividans / Q000235:genetics / Q000378:metabolism
D000900:Anti-Bacterial Agents; D023181:Antimicrobial Cationic Peptides; D001426:Bacterial Proteins; D004269:DNA, Bacterial; D005231:Fatty Acids, Unsaturated; D010455:Peptides; C416817:friulimicin A; C088618:lipase activator protein, Bacteria; D042964:Acyl-CoA Dehydrogenase