Acylated homoserine lactone detection in Pseudomonas aeruginosa biofilms by radiolabel assay.
We describe the development of a new radioactive assay for acyl-HSL production by bacterial cultures. The assay is based on the uptake of radiolabeled methionine and conversion of the radiolabel into SAM. The radiolabeled SAM is then incorporated into acyl-HSL by an acyl-HSL synthase. This assay is faster than previously used bioassays and shows no bias for the detection of acyl-HSLs of a particular length or side chain substitution. Acyl-HSL production can be monitored over a wide range of growth conditions in liquid culture. This assay can also be used in conjunction with a tube biofilm reactor to monitor acyl-HSL production by biofilm cultures. Ultimately this assay will allow comparison of acyl-HSL production by cells subjected to a variety of physiological conditions.
Schaefer
A L
AL
Department of Microbiology, College of Medicine, University of Iowa, Iowa City, Iowa 52242, USA.
Greenberg
E P
EP
Parsek
M R
MR
D015107:4-Butyrolactone / Q000031:analogs & derivatives* / Q000737:chemistry / Q000378:metabolism*; D000215:Acylation; D018441:Biofilms; D001681:Biological Assay; D002250:Carbon Radioisotopes; D015870:Gene Expression; D011550:Pseudomonas aeruginosa / Q000166:cytology / Q000235:genetics / Q000378:metabolism*; D011869:Radioligand Assay / Q000379:methods*
D002250:Carbon Radioisotopes; C088386:homoserine lactone; D015107:4-Butyrolactone