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Comparison of the substrate specificity of two ring-hydroxylating dioxygenases from Sphingomonas sp. VKM B-2434 to polycyclic aromatic hydrocarbons

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Abstract

The genes of two ring-hydroxylating dioxygenases (RHDs) of Sphingomonas sp. VKM B-2434 were cloned and expressed in Escherichia coli. The relative values of the RHD specificity constants were estimated for six polycyclic aromatic hydrocarbons (PAHs) based on the kinetics of PAH mixture conversion by the recombinant strains. The substrate specificity profiles of the enzymes were found to be very different. Dioxygenase ArhA was the most specific to acenaphthylene and showed a low specificity to fluoranthene. Dioxygenase PhnA was the most specific to anthracene and phenanthrene and showed a considerable specificity to fluoranthene. Knockout derivatives of Sphingomonas sp. VKM B-2434 lacking ArhA, PhnA, and both dioxygenases were constructed. PAH degradation by the single-knockout mutants was in agreement with the substrate specificity of the RHD remaining intact. Double-knockout mutant lacking both enzymes was unable to oxidize PAHs. A mutant form of dioxygenase ArhA with altered substrate specificity was described.

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Acknowledgments

The reported study was partially supported by the Russian Foundation for Basic Research (RFBR), Research Project No. 11-04-00831-a.

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Authors and Affiliations

  1. G.K. Skryabin Institute of Biochemistry and Physiology of Microorganisms RAS, Prospekt Nauki 5, Pushchino, Moscow Region, Russia

    Mikhail Baboshin, Tanya Ivashina, Alexey Chernykh & Ludmila Golovleva

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  1. Mikhail Baboshin
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  2. Tanya Ivashina
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  3. Alexey Chernykh
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  4. Ludmila Golovleva
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Correspondence to Ludmila Golovleva.

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Baboshin, M., Ivashina, T., Chernykh, A. et al. Comparison of the substrate specificity of two ring-hydroxylating dioxygenases from Sphingomonas sp. VKM B-2434 to polycyclic aromatic hydrocarbons. Biodegradation 25, 693–703 (2014). https://doi.org/10.1007/s10532-014-9692-3

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  • DOI: https://doi.org/10.1007/s10532-014-9692-3

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