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Associate Professor Hirofumi Ichinose and his research group have constructed a functional library of oxidative enzymes from a mushroom.
Rapid screening of the latent functions of cytochrome P450 monooxygenases in Trametes versicolor
Points
- To understand and utilize the diverse biological functions of mushrooms, it is essential to rapidly identify the functions of cytochrome P450* monooxygenases.
- We successfully constructed a semi-comprehensive functional library of cytochrome P450 enzymes from Trametes versicolor.
- This library is expected to accelerate the elucidation of biosynthetic pathways of valuable natural products, as well as the development of pharmaceuticals and agrochemicals.
Abstract
Mushrooms (basidiomycetes) are eukaryotic microorganisms with significant potential for industrial applications, as they produce bioactive natural products that are relevant to the pharmaceutical field, and are capable of degrading woody biomass and environmental pollutants. This could help achieve a sustainable society. These unique functions are supported by oxidative enzymes known as cytochrome P450s. A typical mushroom encodes between 100-200 genes of P450, and each enzyme exhibits different functions. However, it remains challenging to determine their functions from the genomic information, even in the post-genomic era. Therefore, experimental platforms that enable the rapid and direct identification of P450 functions have to be developed to advance mushroom-based biotechnology.
In this study, we constructed a large-scale functional library of P450 enzymes from the white-rot basidiomycete** Trametes versicolor using yeast. Using the functional library, a large-scale screening and the rapid identification of P450 functions is now possible.
The research group led by Associate Professor Hirofumi Ichinose at the Faculty of Agriculture, Kyushu University, successfully isolated full-length cDNAs for approximately 80% (145 genes) of the P450 encoded in Trametes versicolor. Furthermore, they achieved heterologous expression of these genes in yeast and constructed a semi-comprehensive functional library covering the majority of T. versicolor P450s. They applied this library and identified a P450 enzyme that plays a key role in the biosynthesis of sesquiterpenoids*** for the first time. The successful example contributes to a better understanding of the biosynthesis of valuable natural products.
The functional library developed in this study is highly versatile and can be applied to a wide range of target substrates. It facilitates the discovery of P450 enzymes capable of catalyzing oxidative reactions that are challenging to achieve through conventional organic synthesis, thereby offering promising applications in the industrial synthesis of pharmaceuticals and agrochemicals.
This research was published on April 21, 2025, in the Elsevier journal Enzyme and Microbial Technology.
Researcher's Comment
Through this study, we have established an experimental platform that enables the rapid and direct exploration of latent functions of basidiomycete cytochrome P450 enzymes. We hope that the P450 functional library will serve as a valuable resource for facilitating a wide range of collaborative research efforts.(Hirofumi Ichinose)
Glossary
* Cytochrome P450 (P450):
A class of oxidative enzymes found in a wide range of organisms, responsible for catalyzing various oxidation reactions.
** White-rot basidiomycetes:
A group of fungi capable of degrading lignin in wood, resulting in a bleached appearance. They are highly diverse and widely distributed across the globe.
*** Sesquiterpenoids:
Natural products produced by plants and microorganisms, many of which are known to be bioactive compouns.
Paper Information
Journal: Enzyme Microbial Technology
Title: Functional analysis of cytochrome P450 monooxygenases from the white-rot basidiomycete Trametes versicolor: CYP502B15 decorates cadinane-type sesquiterpenoids
Authors: Hirofumi Ichinose, Megumu Honda, Tsuyoshi Isogai, Chen Chen, Takuya Kitaoka
DOI：10.1016/j.enzmictec.2025.110658
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