A high-throughput assay method to quantify Baeyer-Villiger monooxygenase activity

  • Saß S
  • Kadow M
  • Geitner K
 et al. 
  • 16

    Readers

    Mendeley users who have this article in their library.
  • 6

    Citations

    Citations of this article.

Abstract

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. © 2012 Elsevier Ltd. All rights reserved.

Author-supplied keywords

  • Baeyer-Villiger monooxygenase
  • Biocatalysis
  • HAPMO
  • Spectrophotometric assay
  • Whole cell

Get free article suggestions today

Mendeley saves you time finding and organizing research

Sign up here
Already have an account ?Sign in

Find this document

Authors

  • Stefan Saß

  • Maria Kadow

  • Kristian Geitner

  • Mark L. Thompson

  • Lea Talmann

  • Dominique Böttcher

Cite this document

Choose a citation style from the tabs below

Save time finding and organizing research with Mendeley

Sign up for free