Specific and sensitive plate assay for bacterial lipases.
A plate assay to detect bacterial lipase (EC 3.1.1.3) in a medium containing trioleoylglycerol and the fluorescent dye rhodamine B is presented. Substrate hydrolysis causes the formation of orange fluorescent halos around bacterial colonies visible upon UV irradiation. The logarithm of lipase activity from cell-free culture supernatants is linearly correlated with the diameter of halos, thereby allowing quantitation of lipase activities ranging from 1 to 30 nkat.
D001412:Bacillus subtilis / Q000201:enzymology; D001419:Bacteria / Q000201:enzymology*; D004926:Escherichia coli / Q000201:enzymology; D008049:Lipase / Q000032:analysis*; D000069463:Olive Oil; D010938:Plant Oils; D011550:Pseudomonas aeruginosa / Q000201:enzymology*; D012235:Rhodamines; D012706:Serratia marcescens / Q000201:enzymology; D013211:Staphylococcus aureus / Q000201:enzymology
D000069463:Olive Oil; D010938:Plant Oils; D012235:Rhodamines; D008049:Lipase; C029773:rhodamine B