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Genome-wide identification, annotation and characterization of novel thermostable cytochrome P450 monooxygenases from the thermophilic biomass-degrading fungi Thielavia terrestris and Myceliophthora thermophila

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Abstract

Cytochrome P450 monooxygenases (P450s) are ubiquitous heme-thiolate proteins that have potential biotechnological application. Thermostable-P450s that can withstand hostile industrial conditions, such as high temperatures, extremes of pH and organic solvents, are needed for biotechnological usage. Here, for the first time, we report a large number of thermostable-P450s from two thermophilic biomass-degrading fungi, Myceliophthora thermophila and Thielavia terrestris. Genome-wide P450 analysis revealed the presence of 79 and 70 P450s (P450ome) in T. terrestris and M. thermophila. Authentic P450s containing both the P450 signature domains (EXXR and CXG) were classified as follows: T. terrestris (50 families and 56 subfamilies) and M. thermophila (49 families and 53 subfamilies). Bioinformatics analysis of P450omes suggested the presence of a large number of thermostable-P450s. Based on aliphatic index cut-off (>90), 14 and 11 P450s were determined to be thermostable in T. terrestris and M. thermophila. Among the thermostable P450s, six P450s from T. terrestris and three from M. thermophila had a melting temperature (Tm) of >65 °C, suggesting their hyperthermal tolerance. Analysis of the instability index of two ascomycete P450omes revealed the presence of 12 and 19 in vitro stable P450s in T. terrestris and M. thermophila. Overall, six P450s from T. terrestris and four from M. thermophila showed both thermal tolerance and in vitro stability. Thermophilic ascomycetes P450s are of potential interest from a structural, mechanistic and biotechnological point of view, as five P450s showed higher thermal tolerance and five showed higher in vitro stability compared to the well-characterized thermostable-P450s CYP175A1 (bacteria) and CYP119 (archaea).

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Acknowledgments

Syed thanks the Central University of Technology (CUT) for a grant from the University Research and Innovation Fund. Shale thanks the Medical Research Council (MRC) of South Africa. Krasevec thanks the Slovenian Research Agency Program P1-0104: Functional genomics and biotechnology for health. The authors thank Ms Barbara Bradley, Pretoria, South Africa for English language editing.

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The authors declare that there are no conflict of interest.

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Authors and Affiliations

  1. Department of Life Sciences, Faculty of Health and Environmental Sciences, Central University of Technology, Bloemfontein 9300, Free State, South Africa

    Khajamohiddin Syed & Karabo Shale

  2. Department of Health Sciences, Faculty of Health and Environmental Sciences, Central University of Technology, Bloemfontein 9300, Free State, South Africa

    Samson Sitheni Mashele

  3. School of Basic Studies, Yeungnam University, Gyeongsan, 712-749, South Korea

    K. H. M. Nazmul Hussain Nazir

  4. National Institute of Chemistry, Hajdrihova 19, 1000, Ljubljana, Slovenia

    Nada Krasevec

  5. Department of Chemical Engineering, University of Alberta, Edmonton, AB, Canada

    Nataraj Sekhar Pagadala

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  1. Khajamohiddin Syed
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Correspondence to Khajamohiddin Syed.

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Syed, K., Shale, K., Nazir, K.H.M.N.H. et al. Genome-wide identification, annotation and characterization of novel thermostable cytochrome P450 monooxygenases from the thermophilic biomass-degrading fungi Thielavia terrestris and Myceliophthora thermophila . Genes Genom 36, 321–333 (2014). https://doi.org/10.1007/s13258-013-0170-9

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