Artículos de revistas
A fluorescence-based assay for Baeyer-Villiger monooxygenases, hydroxylases and lactonases
Registro en:
Advanced Synthesis & Catalysis. Wiley-blackwell, v. 347, n. 41858, n. 1041, n. 1050, 2005.
1615-4150
WOS:000229760500019
10.1002/adsc.200505040
Autor
Sicard, R
Chen, LS
Marsaioli, AJ
Reymond, JL
Institución
Resumen
Alkylation of umbelliferone and nitrophenol with chloroacetone, 3-chlorobutanone, 2-chlorocyclopentanone and 2-chlorocyclohexanone gave the corresponding 2-coumaryloxy and 2-nitrophenoxy ketones. The 2-coumarytoxy ketones were used as fluorogenic substrates to detect Baeyer-Villiger monooxygenases activities of microbial cultures in high-throughput using microtiter plates. The 2-coumaryloxy ketones were oxidized by microorganisms producing Baeyer-Villiger monooxygenases (BVMO), releasing umbelliferone as a fluorescent signal. The substrates were also biotransformed by a microbial monooxygenase (Trichosporon cutaneum). Chemical Baeyer-Villiger oxidation of 2-coumaryloxy ketones using meta-chloroperbenzoic acid proceeded regioselectively to the corresponding acyloxyalkyl derivatives of umbelliferone and nitrophenol. These chiral lactones underwent a fluorogenic and chromogenic reaction upon hydrolysis by esterases, in particular pig liver esterase. Enantioselectivity of the ester hydrolysis reaction was determined by chiral-phase analysis of the unreacted lactones. 347 41858 1041 1050