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

다국어 초록 (Multilingual Abstract)

Cytochrome P450 monooxygenases (P450s)are ubiquitous heme-thiolate proteins that have potentialbiotechnological application. Thermostable-P450s that canwithstand hostile industrial conditions, such as high temperatures,extremes of pH and organic solvents, are neededfor biotechnological usage. Here, for the first time, wereport a large number of thermostable-P450s from twothermophilic biomass-degrading fungi, Myceliophthorathermophila and Thielavia terrestris. Genome-wide P450analysis revealed the presence of 79 and 70 P450s(P450ome) in T. terrestris and M. thermophila. AuthenticP450s containing both the P450 signature domains (EXXRand CXG) were classified as follows: T. terrestris (50families and 56 subfamilies) and M. thermophila (49families and 53 subfamilies). Bioinformatics analysis ofP450omes suggested the presence of a large number ofthermostable-P450s. Based on aliphatic index cut-off([90), 14 and 11 P450s were determined to be thermostablein T. terrestris and M. thermophila. Among thethermostable P450s, six P450s from T. terrestris and threefrom M. thermophila had a melting temperature (Tm) of[65 C, suggesting their hyperthermal tolerance. Analysisof the instability index of two ascomycete P450omesrevealed the presence of 12 and 19 in vitro stable P450s inT. terrestris and M. thermophila. Overall, six P450s fromT. terrestris and four from M. thermophila showed boththermal tolerance and in vitro stability. Thermophilicascomycetes P450s are of potential interest from a structural,mechanistic and biotechnological point of view, asfive P450s showed higher thermal tolerance and fiveshowed higher in vitro stability compared to the wellcharacterizedthermostable-P450s CYP175A1 (bacteria)and CYP119 (archaea).

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