An improved procedure for the purification of catalytically active alkane hydroxylase from Pseudomonas putida GPo1

Meng Xie, Hernan Alonso, Anna Roujeinikova

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7 Citations (Scopus)

Abstract

Bacterial alkane hydroxylases are of high interest for bioremediation applications as they allow some bacteria to grow in oil-contaminated environments. Furthermore, they have tremendous biotechnological potential as they catalyse the stereo- and regio-specific hydroxylation of chemically inert alkanes, which can then be used in the synthesis of pharmaceuticals and other high-cost chemicals. Despite their potential, progress on the detailed characterization of these systems has so far been slow mainly due to the lack of a robust procedure to purify its membrane protein component, monooxygenase AlkB, in a stable and active form. This study reports a new method for isolating milligramme amounts of recombinant Pseudomonas putida GPo1 AlkB in a folded, catalytically active form to purity levels above 90%. AlkB solubilised and purified in the detergent lauryldimethylamine oxide was demonstrated to be active in catalysing the epoxidation reaction of 1-octene with an estimated Km value of 0.2 mM.
Original languageEnglish
Pages (from-to)823-831
Number of pages9
JournalApplied Biochemistry and Biotechnology - Part A Enzyme Engineering and Biotechnology
Volume165
Issue number3
DOIs
Publication statusPublished - Oct 2011

Keywords

  • biocatalysis
  • alkane hydroxylase
  • biomodification of hydrocarbons
  • alkane biodegradation

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