title: Bacillus megaterium CYP102A1 oxidation of acyl homoserine lactones and acyl homoserines.
abstract: Quorum sensing, the ability of bacteria to sense their own population density through the synthesis and detection of small molecule signals, has received a great deal of attention in recent years. Acyl homoserine lactones (AHLs) are a major class of quorum sensing signaling molecules. In nature, some bacteria that do not synthesize AHLs themselves have developed the ability to degrade these compounds by cleaving the amide bond or the lactone ring. By inactivating this signal used by competing bacteria, the degrading microbe is believed to gain a competitive advantage. In this work we report that CYP102A1, a widely studied cytochrome P450 from Bacillus megaterium, is capable of very efficient oxidation of AHLs and their lactonolysis products acyl homoserines. The previously known substrates for this enzyme, fatty acids, can also be formed in nature by hydrolysis of the amide of AHLs, so CYP102A1 is capable of inactivating the active parent compound and the products of both known pathways for AHL inactivation observed in nature. AHL oxidation primarily takes place at the omega-1, omega-2, and omega-3 carbons of the acyl chain, similar to this enzyme's well-known activity on fatty acids. Acyl homoserines and their lactones are better substrates for CYP102A1 than fatty acids. Bioassay of the quorum sensing activity of oxidation products reveals that the subterminally hydroxylated AHLs exhibit quorum sensing activity, but are 18-fold less active than the parent compound. In vivo, B. megaterium inactivates AHLs by a CYP102A1 dependent mechanism that must involve additional components that further sequester or metabolize the products, eliminating their quorum sensing activity. Cytochrome P450 oxidation of AHLs represents an important new mechanism of quorum quenching.
authors.affiliation: Department of Chemistry, University of Texas at Dallas, Richardson, Texas 75080, USA.
authors.forename: Puneet K
authors.initials: PK
authors.lastname: Chowdhary
authors.forename: Neela
authors.initials: N
authors.lastname: Keshavan
authors.forename: Hien Q
authors.initials: HQ
authors.lastname: Nguyen
authors.forename: Julian A
authors.initials: JA
authors.lastname: Peterson
authors.forename: Juan E
authors.initials: JE
authors.lastname: González
authors.forename: Donovan C
authors.initials: DC
authors.lastname: Haines
mesh_terms: D054742:Acyl-Butyrolactones / Q000737:chemistry / Q000378:metabolism*; D001410:Bacillus megaterium / Q000201:enzymology / Q000378:metabolism*; D001426:Bacterial Proteins / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism*; D008401:Gas Chromatography-Mass Spectrometry; D006714:Homoserine / Q000737:chemistry / Q000378:metabolism*; D007700:Kinetics; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D015394:Molecular Structure; D009251:NADPH-Ferrihemoprotein Reductase; D010084:Oxidation-Reduction; D011485:Protein Binding; D053038:Quorum Sensing
chemical_list: D054742:Acyl-Butyrolactones; D001426:Bacterial Proteins; D006714:Homoserine; D003577:Cytochrome P-450 Enzyme System; D006899:Mixed Function Oxygenases; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases