Identification and exploitation of biosynthetic pathways from Ascocoryne sarcoides

真菌Ascocoryne sarcoides能产生多种挥发性有机化合物(VOCs)，包括烷烃、支链醇类、酮类、Esthers等。这些VOCs是典型的低分子代谢物，由于它们作为燃料替代品、平台化学品和香气成分的潜在用途而引起人们的极大兴趣。然而，许多导致真菌中VOC生物合成的代谢途径仍然未知，这阻碍了它们在生物技术应用中的开发。已知的五个密切相关的结节杆菌分离株产生不同的VOC谱。我们已经对其中四个分离株的基因组进行了测序，第五个是公开可用的。我们将把我们的基因组知识与生化分析(以及适当的转录和/或蛋白质组学分析)相结合，以阐明与工业相关的生物合成途径。候选的生物合成途径将转移到合适的微生物宿主上。这提供了三个好处：i)代谢途径的重建提供了正确识别基因(S)的证据；ii)它允许对基因产物(S)进行进一步的表征；iii)这些技术可以通过候选基因的组合表达来识别未知的途径。DNA合成和文库组装方法极大地促进了这一点，从而消除了传统克隆的需要。

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.