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A novel class of fungal lipoxygenases

  • Applied genetics and molecular biotechnology
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Abstract

Lipoxygenases (LOXs) are well-studied enzymes in plants and mammals. However, fungal LOXs are less studied. In this study, we have compared fungal LOX protein sequences to all known characterized LOXs. For this, a script was written using Shell commands to extract sequences from the NCBI database and to align the sequences obtained using Multiple Sequence Comparison by Log-Expectation. We constructed a phylogenetic tree with the use of Quicktree to visualize the relation of fungal LOXs towards other LOXs. These sequences were analyzed with respect to the signal sequence, C-terminal amino acid, the stereochemistry of the formed oxylipin, and the metal ion cofactor usage. This study shows fungal LOXs are divided into two groups, the Ile- and the Val-groups. The Ile-group has a conserved WRYAK sequence that appears to be characteristic for fungal LOXs and has as a C-terminal amino acid Ile. The Val-group has a highly conserved WL-L/F-AK sequence that is also found in LOXs of plant and animal origin. We found that fungal LOXs with this conserved sequence have a Val at the C-terminus in contrast to other LOXs of fungal origin. Also, these LOXs have signal sequences implying these LOXs will be expressed extracellularly. Our results show that in this group, in addition to the Gaeumannomyces graminis and the Magnaporthe salvinii LOXs, the Aspergillus fumigatus LOX uses manganese as a cofactor.

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Acknowledgments

We would like to thank Dr. Ir. P.J. Schaap for his help writing the scripts used in this study, Ing. P. R. Nobels for his help with the ICP-AES, and C. Nebel for her help with the HPLC experiment. The authors gratefully acknowledge the financial support provided by the European Research Project (Novel enzyme tools for production of functional oleochemicals from unsaturated lipids (ERA-NOEL), ERA-IB/BIO/0001/2008).

Conflict of interest

The authors declare that they have no competing interests.

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Authors and Affiliations

  1. Laboratory of Systems and Synthetic Biology, Microbial Systems Biology, Wageningen University, Dreijenplein 10, 6703 HB, Wageningen, The Netherlands

    Ruud Heshof & Leo H. de Graaff

  2. VTT Biotechnology, P.O. Box 1500, 02044 VTT, Espoo, Finland

    Sirpa Jylhä

  3. AB Enzymes GmbH, Feldbergstrasse 78, 64293, Darmstadt, Germany

    Thomas Haarmann

  4. Faculty of Science and Technology, Department of Food Science, Aarhus University, Blichers Allé 20, P.O. Box 50, 8830, Tjele, Denmark

    Ann Louise Worsøe Jørgensen & Trine Kastrup Dalsgaard

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  1. Ruud Heshof
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  2. Sirpa Jylhä
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  4. Ann Louise Worsøe Jørgensen
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  5. Trine Kastrup Dalsgaard
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  6. Leo H. de Graaff
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Correspondence to Leo H. de Graaff.

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Heshof, R., Jylhä, S., Haarmann, T. et al. A novel class of fungal lipoxygenases. Appl Microbiol Biotechnol 98, 1261–1270 (2014). https://doi.org/10.1007/s00253-013-5392-x

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  • DOI: https://doi.org/10.1007/s00253-013-5392-x

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