Metabolism of chlorobiphenyls by a variant biphenyl dioxygenase exhibiting enhanced activity toward dibenzofuran

Biochemical and Biophysical Research Communications
Jean-François VigerM Sylvestre

Abstract

The biphenyl dioxygenase of Burkholderia xenovorans LB400 (BphAE(LB400)) catalyzes the dihydroxylation of biphenyl and of several polychlorinated biphenyls (PCBs) but it poorly oxidizes dibenzofuran. In this work we showed that BphAE(RR41), a variant which was previously found to metabolize dibenzofuran more efficiently than its parent BphAE(LB400), metabolized a broader range of PCBs than BphAE(LB400). Hence, BphAE(RR41) was able to metabolize 2,6,2',6'-, 3,4,3',5'- and 2,4,3',4'-tetrachlorobiphenyl that BphAE(LB400) is unable to metabolize. BphAE(RR41) was obtained by changing Thr335Phe336Asn338Ile341Leu409 of BphAE(LB400) to Ala335Met336Gln338Val341Phe409. Site-directed mutagenesis was used to create combinations of each substitution, in order to assess their individual contributions. Data show that the same Asn338Glu/Leu409Phe substitution that enhanced the ability to metabolize dibenzofuran resulted in a broadening of the PCB substrates range of the enzyme. The role of these substitutions on regiospecificities toward selected PCBs is also discussed.

References

Jul 9, 1999·Biotechnology and Bioengineering·F Brühlmann, W Chen
Jun 26, 2002·Journal of Bacteriology·Diane BarriaultMichel Sylvestre
Sep 6, 2003·Applied and Environmental Microbiology·Michael SeegerSilvia Sepúlveda-Boza
Aug 5, 2004·Journal of Bacteriology·Kensuke FurukawaMasatoshi Goto
Sep 20, 2005·Biochemical and Biophysical Research Communications·Daniel J FerraroS Ramaswamy
Jan 5, 2006·Organic & Biomolecular Chemistry·Derek R Boyd, Timothy D H Bugg
Jun 10, 2011·The Journal of Biological Chemistry·Mahmood MohammadiMichel Sylvestre

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Citations

May 2, 2012·Biochemical and Biophysical Research Communications·Pravindra KumarMichel Sylvestre
Oct 27, 2012·Current Opinion in Biotechnology·Ana Segura, Juan Luis Ramos
Jul 28, 2020·Applied and Environmental Microbiology·Junde LiXiaoke Hu

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