Acylhomoserine lactone synthase activity of the Vibrio fischeri AinS protein.
Acylhomoserine lactones, which serve as quorum-sensing signals in gram-negative bacteria, are produced by members of the LuxI family of synthases. LuxI is a Vibrio fischeri enzyme that catalyzes the synthesis of N-(3-oxohexanoyl)-L-homoserine lactone from an acyl-acyl carrier protein and S-adenosylmethionine. Another V. fischeri gene, ainS, directs the synthesis of N-octanoylhomoserine lactone. The AinS protein shows no significant sequence similarity with LuxI family members, but it does show sequence similarity with the Vibrio harveyi LuxM protein. The luxM gene is required for the synthesis of N-(3-hydroxybutyryl)-L-homoserine lactone. To gain insights about whether AinS and LuxM represent a second family of acylhomoserine lactone synthases, we have purified AinS as a maltose-binding protein (MBP) fusion protein. The purified MBP-AinS fusion protein catalyzed the synthesis of N-octanoylhomoserine lactone from S-adenosylmethionine and either octanoyl-acyl carrier protein or, to a lesser extent, octanoyl coenzyme A. With the exception that octanoyl coenzyme A served as an acyl substrate for the MBP-AinS fusion protein, the substrates for and reaction kinetics of the MBP-AinS fusion protein were similar to those of the several LuxI family members previously studied. We conclude that AinS is an acylhomoserine lactone synthase and that it represents a second family of such enzymes.
Hanzelka
B L
BL
Department of Microbiology, University of Iowa, Iowa City, Iowa 52242, USA.
Parsek
M R
MR
Val
D L
DL
Dunlap
P V
PV
Cronan
J E
JE
Greenberg
E P
EP
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism*; D007700:Kinetics; D007783:Lactones / Q000378:metabolism*; D008956:Models, Chemical; D011993:Recombinant Fusion Proteins / Q000302:isolation & purification / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D013379:Substrate Specificity; D014733:Vibrio / Q000235:genetics / Q000378:metabolism*
C096878:AinS protein, Vibrio fischeri; D001426:Bacterial Proteins; D007783:Lactones; D011993:Recombinant Fusion Proteins; C084288:luxM protein, Vibrio harveyi; D012436:S-Adenosylmethionine