Identification and exploitation of biosynthetic pathways from Ascocoryne sarcoides

The fungi Ascocoryne sarcoides produce a diverse and extensive range of volatile organic compounds (VOCs), including alkanes, branched-chain alcohols, ketones, esthers. These VOCs are typically low molecular weight metabolites and are of considerable interest be-cause of their potential use as fuel substitutes, platform chemicals and aroma components. However, many of the metabolic pathways leading to VOC biosynthesis in fungi remain un-known, preventing their exploitation in biotechnological applications.

Five closely related A. sarcoides isolates are known to generate different VOC profiles. We have sequenced the genomes of four of these isolates, the fifth is publically available. We will combine our genomic knowledge with biochemical analysis (and where appropriate transcriptional and/or proteomic analysis) to elucidate biosynthetic pathways of industrial relevance. Candidate biosynthetic pathways will be transferred to a suitable microbial host. This provides three benefits: i) reconstitution of metabolic pathways provides proof that gene(s) have been correctly identified; ii) it allows further characterisation of the gene product(s); and iii) these technologies can be leveraged to identify obscure pathways through combinatorial expression of candidate genes. This is greatly facilitated by DNA syn-thesis and library assembly methodologies obviating the need for traditional cloning.