A method for rapid screening of ketone biotransformations: detection of whole cell Baeyer-Villiger monooxygenase activity.
A method for screening of ketone biotransformations was developed and applied to the identification of Baeyer-Villiger monooxygenase (BVMO) activity. The method was based on the formation of a purple coloured product on reaction between an enolizable ketone and 3,5-dinitrobenzoic acid in an alkaline solution. Absorbance of the colour decreased with the size of the cycloketone ring. Stoichiometric ratio between cycloketone and 3,5-dinitrobenzoic acid was 1:1 at maximum absorbance. The method was applied for monitoring the consumption of cyclohexanone by bacteria under aerobic conditions, and was found to be potentially useful for both screening assays and quantitative measurements of BVMO activity. Compared to other existing methods, this method is faster, cheaper and amenable for whole cell assays.
Department of Biotechnology, Center for Chemistry & Chemical Engineering, Lund University, Box 124, SE-221 00 Lund, Sweden.
Javier A
JA
Linares-Pastén
Georgina
G
Chávez-Lizárraga
Rodrigo
R
Villagomez
Gashaw
G
Mamo
Rajni
R
Hatti-Kaul
D001419:Bacteria / Q000166:cytology / Q000201:enzymology* / Q000254:growth & development; D001709:Biotechnology / Q000379:methods*; D001711:Biotransformation; D002849:Chromatography, Gas; D002851:Chromatography, High Pressure Liquid; D003124:Colorimetry; D003512:Cyclohexanones / Q000737:chemistry / Q000378:metabolism; D007659:Ketones / Q000378:metabolism*; D006899:Mixed Function Oxygenases / Q000378:metabolism*; D009579:Nitrobenzoates / Q000378:metabolism; D010084:Oxidation-Reduction; D013379:Substrate Specificity; D013997:Time Factors
D003512:Cyclohexanones; D007659:Ketones; D009579:Nitrobenzoates; C026017:3,5-dinitrobenzoic acid; C036468:cyclohexanone; D006899:Mixed Function Oxygenases