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Please use this identifier to cite or link to this item: http://repository.iitr.ac.in/handle/123456789/26949
Title: Structural insights into the metabolism of 2-chlorodibenzofuran by an evolved biphenyl dioxygenase
Authors: Kumar, Pravindra R.Manish
Mohammadi M.
Dhindwal S.
Pham T.T.M.
Bolin J.T.
Sylvestre M.
Published in: Biochemical and Biophysical Research Communications
Abstract: The biphenyl dioxygenase of Burkholderia xenovorans LB400 (BphAE LB400) is a Rieske-type oxygenase that catalyzes the stereospecific oxygenation of many heterocyclic aromatics including dibenzofuran. In a previous work, we evolved BphAE LB400 and obtained BphAE RR41. This variant metabolizes dibenzofuran and 2-chlorodibenzofuran more efficiently than BphAE LB400. However, the regiospecificity of BphAE RR41 toward these substrates differs. Dibenzofuran is metabolized principally through a lateral dioxygenation whereas 2-chlorodibenzofuran is metabolized principally through an angular dioxygenation. In order to explain this difference, we examined the crystal structures of both substrate-bound forms of BphAE RR41 obtained under anaerobic conditions. This structure analysis, in combination with biochemical data for a Ser283Gly mutant provided evidences that the substrate is compelled to move after oxygen-binding in BphAE RR41:dibenzofuran. In BphAE RR41:2-chlorodibenzofuran, the chlorine atom is close to the side chain of Ser283. This contact is missing in the BphAE RR41:dibenzofuran, and strong enough in the BphAE RR41:2-chlorodibenzofuran to help prevent substrate movement during the catalytic reaction. © 2012 Elsevier Inc.
Citation: Biochemical and Biophysical Research Communications, 421(4): 757-762
URI: https://doi.org/10.1016/j.bbrc.2012.04.078
http://repository.iitr.ac.in/handle/123456789/26949
Issue Date: 2012
Keywords: Biocatalysis
Burkholderia xenovorans LB400
Chlorodibenzofurans
Directed evolution
Enzyme engineering
Rieske-type oxygenase
ISSN: 0006291X
Author Scopus IDs: 55064809000
23390544500
36082537700
54791518500
57197844041
7005984229
Author Affiliations: Kumar, P., Department of Biological Sciences and Center for Cancer Research, Purdue University, West Lafayette, IN 47907, United States, Department of Biotechnology, Indian Institute of Technology, Roorkee 247667, India
Mohammadi, M., Institut National de la Recherche Scientifique, INRS-Institut Armand-Frappier, Laval, QC, H7V 1B7, Canada, Département de génie informatique et génie logiciel, École Polytechnique de Montréal, C.P. 6079, succ. Centre-ville, Montreal, QC, H3C 3A7, Canada
Dhindwal, S., Department of Biotechnology, Indian Institute of Technology, Roorkee 247667, India
Pham, T.T.M., Institut National de la Recherche Scientifique, INRS-Institut Armand-Frappier, Laval, QC, H7V 1B7, Canada
Bolin, J.T., Department of Biological Sciences and Center for Cancer Research, Purdue University, West Lafayette, IN 47907, United States
Sylvestre, M., Institut National de la Recherche Scientifique, INRS-Institut Armand-Frappier, Laval, QC, H7V 1B7, Canada
Funding Details: This work was supported by the Natural Sciences and Engineering Research Council of Canada (NSERC) (Grant # RGPIN/39579-2007 ). X-ray diffraction data were collected at APS using Southeast Regional Collaborative Access Team (SER-CAT) 22-ID beamline; supporting institutions may be found at http://ser-cat.org/members.html . PK thanks the DRDO, India for providing the financial support and also thank MCU facility at IIC, IIT Roorkee for carrying out structure determination and analysis. Natural Sciences and Engineering Research Council of Canada, NSERC: RGPIN/39579-2007; Defence Research and Development Organisation, DRDO
Corresponding Author: Sylvestre, M.; Institut National de la Recherche Scientifique (INRS-Institut Armand-Frappier), Laval, QC, H7V 1B7, Canada; email: Michel.Sylvestre@iaf.inrs.ca
Appears in Collections:Journal Publications [BT]

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