کد مقاله کد نشریه سال انتشار مقاله انگلیسی نسخه تمام متن
5218516 1383330 2012 6 صفحه PDF دانلود رایگان
عنوان انگلیسی مقاله ISI
A high-throughput assay method to quantify Baeyer-Villiger monooxygenase activity
موضوعات مرتبط
مهندسی و علوم پایه شیمی شیمی آلی
پیش نمایش صفحه اول مقاله
A high-throughput assay method to quantify Baeyer-Villiger monooxygenase activity
چکیده انگلیسی

An assay for the spectrophotometric determination of Baeyer-Villiger monooxygenase (BVMO) activity is described. Baeyer-Villiger oxidation of p-nitroacetophenone generates the corresponding acetate and subsequent hydrolysis of this ester by an esterase or NaOH results in the formation of p-nitrophenolate. This chromophore can be easily quantified spectrophotometrically at 410 nm. The assay can be performed in a microtiter plate format and is applicable to whole Escherichia coli cells containing recombinant BVMO, crude cell extract as well as using purified enzyme as exemplified for the 4-hydroxyacetophenone monooxygenase (HAPMO) from Pseudomonas putida JD1. Furthermore, the assay was used to identify more active HAPMO variants within enzyme mutant libraries generated by error-prone PCR or site-saturation mutagenesis.

ناشر
Database: Elsevier - ScienceDirect (ساینس دایرکت)
Journal: Tetrahedron - Volume 68, Issue 37, 16 September 2012, Pages 7575-7580
نویسندگان
, , , , , , , ,