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- Title
An Improved Procedure for the Purification of Catalytically Active Alkane Hydroxylase from Pseudomonas putida GPo1.
- Authors
Xie, Meng; Alonso, Hernan; Roujeinikova, Anna
- 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 K value of 0.2 mM.
- Subjects
ALKANES; PSEUDOMONAS; ENZYMES; HYDROCARBONS; BIODEGRADATION
- Publication
Applied Biochemistry & Biotechnology, 2011, Vol 165, Issue 3/4, p823
- ISSN
0273-2289
- Publication type
Article
- DOI
10.1007/s12010-011-9300-5