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.
► Development of a rapid screening assay for biotransformation of cycloketones. ► Assay validated for screening of Baeyer–Villiger monooxygenase activity in whole cells. ► Assay compared with the known GC and HPLC methods for determination of BVMO activity.
Journal: Enzyme and Microbial Technology - Volume 50, Issue 2, 10 February 2012, Pages 101–106