A single mutation in P450BM-3 enhances acyl homoserine lactone: acyl homoserine substrate binding selectivity nearly 250-fold.
Quorum sensing is the process by which bacteria alter gene regulation in response to their population density. The enzymatic inactivation of quorum signals has shown promise for use in genetically modified organisms resistant to pathogens. We recently characterized the ability of a cytochrome P450, P450BM-3, to oxidize the quorum sensing signals known as acyl homoserine lactones. The oxidation of the acyl homoserine lactones reduced their activity as quorum signals. The enzyme also oxidized the inactive lactonolysis products, acyl homoserines. The enzyme showed similar binding affinity for the acyl homoserine lactones and acyl homoserines. The latter reaction may lead to problems when lactonases and the P450-dependent system are used in tandem, as oxidation of the acyl homoserines produced by lactonolysis in vivo may compete with acyl homoserine lactone oxidation by the cytochrome P450. We report here that a single mutation (R47S) in P450BM-3 is capable of increasing the acyl homoserine lactone: acyl homoserine substrate binding selectivity of the enzyme nearly 250-fold, reducing the potential for competition by acyl homoserines and significantly enhancing the potential for use of P450BM-3 as part of a pathogen resistance system in genetically modified crops.
Chowdhary
Puneet K
PK
Department of Chemistry, University of Texas at Dallas, Richardson, TX 75080, United States.
Stewart
Larissa
L
Lopez
Claudia
C
Haines
Donovan C
DC
D054742:Acyl-Butyrolactones / Q000378:metabolism*; D001120:Arginine; D001410:Bacillus megaterium / Q000201:enzymology*; D001426:Bacterial Proteins / Q000235:genetics* / Q000378:metabolism*; D003577:Cytochrome P-450 Enzyme System / Q000235:genetics* / Q000378:metabolism*; D005998:Glycine / Q000031:analogs & derivatives / Q000737:chemistry; D006714:Homoserine / Q000378:metabolism*; D007700:Kinetics; D009154:Mutation / Q000235:genetics*; D009251:NADPH-Ferrihemoprotein Reductase / Q000235:genetics* / Q000378:metabolism*; D010169:Palmitic Acids / Q000737:chemistry; D013056:Spectrophotometry, Ultraviolet; D013379:Substrate Specificity
D054742:Acyl-Butyrolactones; D001426:Bacterial Proteins; D010169:Palmitic Acids; C095773:N-palmitoylglycine; D006714:Homoserine; D003577:Cytochrome P-450 Enzyme System; D001120:Arginine; D009251:NADPH-Ferrihemoprotein Reductase; C052423:flavocytochrome P450 BM3 monoxygenases; D005998:Glycine