Accessing the hidden biosynthetic capabilities of fungi

摘要 丝状真菌产生大量具有生物活性的次生代谢物 这对医学和农业的进步至关重要。例如，真菌短信 表现出细胞抑制、免疫抑制、降脂或抗菌特性。快速发展 在丝状真菌的基因组测序中，发现了非常大量的推测 没有已知代谢物的生物合成途径，表明发现 新的化合物和活性。然而，对真菌充分表征的重大障碍 BGC多样性存在：(A)许多Sms是由“非规范的”生物合成基因簇合成的 生物信息学算法无法识别(BGC)，(B)许多BGC未在标准中表达 (C)某些生物合成途径的基因不是 所有基因都聚集在一起，涉及一个以上的基因座。此外，(D)对 功能性BGC的起源。我们最近的研究结果清楚地表明，非正则大气球揭示了 真正新颖的结构或不寻常的生物化学，这种特定的真菌分化信号 诱导全球BGC表达，重组热点产生BGC多样性。在……里面 这笔赠款，我们将(I)表征异氰化物合成酶(ICS)BGC，一种最近发现的 未被当前软件算法识别的非规范真菌BGC家族 有新的和令人兴奋的生物化学的初步结果，(Ii)使用真菌 差异信号用于转录识别不同来源的“隐形”BGC 真菌分类群和(Iii)解决了基因组重组和重组热点的假设 横向转移催生了新的BGC。基于非正则基因的前提 团簇特别可能产生高度结构和结构的化学实体 功能新颖性与两项先进技术相结合，能够识别活跃的BGC以及BGC如何 形成了我们的比较转录学工具集，高级内源和异源 表达平台和最近开发的比较代谢组学平台将 提供丰富的新结构、生物合成途径和生物活性 真菌BGC的生物学、遗传学和化学研究的拓展。此外，对BGC的洞察 全球BGC诱导信号的产生和鉴定呈现出真正新的过程 进一步介绍了真菌BGC的发现范围和功能注释。

ABSTRACT Filamentous fungi produce a vast universe of secondary metabolites (SM) with biological activities that are of central importance for progress in medicine and agriculture. For example, fungal SMs exhibit cytostatic, immunosuppressant, lipid lowering, or antimicrobial properties. Rapid progress in sequencing the genomes of filamentous fungi has revealed a very large number of putative biosynthetic pathways with no known metabolites, suggesting a vast potential for the discovery of new compounds and activities. However, significant impediments to full characterization of fungal BGC diversity exist: (a) many SMs are synthesized by ‘non-canonical’ biosynthetic gene clusters (BGCs) not recognized by bioinformatic algorithms, (b) many BGCs are not expressed in standard laboratory conditions (e.g. ‘cryptic’ BGCs), and (c) genes for some biosynthetic pathways are not all clustered and involve more than one locus. Further, there is (d) little understanding of the genesis of functional BGCs. Our recent results clearly indicate that non-canonical BGCs reveal genuinely novel structures or unusual biochemistry, that specific fungal differentiation signals induce global BGC expression and that “hot spots” of recombination generate BGC diversity. In this grant, we will (i) characterize isocyanide synthase (ICS) BGCs, a recently discovered family of noncanonical fungal BGCs not recognized by current software algorithms for which we have preliminary results demonstrating new and exciting biochemistry, (ii) use a fungal differentiation signal for transcriptomic identification of ‘invisible' BGCs across diverse fungal taxa and (iii) address the hypothesis that genomic “hot spots” of recombination and horizontal transfer give birth to new BGCs. Based on the premise that non-canonical gene clusters are particularly likely to produce chemical entities with a high degree of structural and functional novelty coupled with two advances able to identify active BGCs and how BGC are formed, our tool set of comparative transcriptomics, advanced endogenous and heterologous expression platforms and a recently developed platform for comparative metabolomics will provide a wealth of new structures, biosynthetic pathways, and biological activities through expansion of the biology, genetics and chemistry of fungal BGCs. Moreover, insight into BGC genesis and identification of global BGC induction signals present genuinely new processes to further the scope of fungal BGC discovery and functional annotation.