Biotransformation of Androstenedione by Filamentous Fungi Isolated from Cultural Heritage Sites in the State Tretyakov Gallery

SIMPLE SUMMARY: Microorganisms are able to grow on substrates of the most diverse nature. One of the most practical habitats, in terms of cultural heritage conservation, is fine art objects such as tempera or oil paintings on canvas. Since tempera paints are produced on the basis of egg yolk, which is one of the richest sources of cholesterol in nature (up to 2% of dry weight), and in the process of aging of tempera materials, changes in cholesterol do not affect the core structure of the steroid nucleus, the group of fungi that we have isolated are tempera painting destructors is seen as a promising object for screening for their possible steroid-transforming activities. In this regard, the purpose of our work was to determine the ability to transform pharmaceutically significant steroids with dominant fungi-destructors of tempera paintings, previously isolated in the State Tretyakov Gallery. Consequently, we have demonstrated for the first time that fungi-destructors of tempera paintings have steroid-transforming activity and are promising microorganisms for screening for biotechnologically significant transformations of steroids with further industrial use. ABSTRACT: The transformation of steroids by microorganisms is widely used in medical biotechnology. A huge group of filamentous fungi is one of the most promising taxa for screening new biocatalytic reactions in order to obtain pharmaceutically significant steroids. In this work, we screened 10 filamentous fungi-destructors of egg tempera for the ability to biotransform androst-4-en-3,17-dione (AD) during cultivation in a liquid nutrient medium or in a buffer solution. These taxonomically unrelated strains, belonging to the classes Eurotiomycetes, Dothideomycetes and Sordariomycetes, are dominant representatives of the microbiome from halls where works of tempera painting are stored in the State Tretyakov Gallery (STG, Moscow, Russia). Since the binder of tempera paints, egg yolk, contains about 2% cholesterol, these degrading fungi appear to be a promising group for screening for steroid converting activity. It turned out that all the studied fungi-destructors are able to transform AD. Some strains showed transformation efficiency close to the industrial strain Curvularia lunata RNCIM F-981. In total, 33 steroids formed during the transformation of AD were characterized, for 19 of them the structure was established by gas chromatography/mass spectrometry analysis. In this work, we have shown for the first time that fungi-destructors of tempera paintings can efficiently transform steroids.