An endophytic Hypoxylon sp. (BS15) has recently been found to produce volatile organic compounds (VOCs) that have potential relevance as hydrocarbon fuels (“mycodiesel”). Traditionally, a refined carbohydrate (e.g., sucrose) was required as a diet source. Recently, a solvent free green chemistry mechanocatalytic method involving ball milling in the solid state has been used to convert cellulose waste products to simple carbohydrate. The growth of BS15 was obtained on this degraded cellulose as well as the more traditional potato dextrose broth. The volatile compounds produced from both media were largely the same. Solid-phase extraction was used to extract BS15 medium in order to reduce the solvent waste and more efficiently retain compounds having low vapor pressures, which retained a significantly larger variety of the volatile compounds than the more usual liquid–liquid extraction or solid-phase micro-extraction. Unfortunately, it is observed that long term in vitro growth of the fungus results in diminished VOC production. The VOC production was partially restored by cultivating BS15 in growth media containing finely ground woody tissue from the original host plant (Taxodium distichum). Extracts from this woody tissue were made by sequentially extracting with dichloromethane, methanol, and water with a goal of isolating VOC production modulators. Both the dichloromethane and water extracts place on bio-mimicking filter paper were found to modulate VOC production, while the methanol extract had no significant impact. Surprisingly, the woody tissue remaining after exhaustive extraction also acted as a VOC production modulator when combined with the growth media, with noticeable changes in the production of four compounds. This woody tissue also induced production of two compounds not observed in the original BS15 extract. Remarkably filter paper had the same modulating effect as exhaustively extracted woody tissue, suggesting the modulation was partially due to cellulose degradation products. As a result, this study demonstrated that VOC production in BS15 can be influenced by multiple compounds in the woody tissue rather than a single modulator.

Isolation of endophytic fungus Hypoxylon sp. BS15 producing volatile organic compounds (VOCs) with fuel potential and epigenetically altering VOC production over time

2019 Spring National Meeting

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