School of Life Sciences, University of Warwick, Coventry, CV4 7AL, UK.
Department of Chemistry, University of Warwick, Coventry, CV4 7AL, UK.
BMC Genomics. 2024 Nov 26;25(1):1141. doi: 10.1186/s12864-024-11064-w.
Fungi are talented producers of secondary metabolites with applications in the pharmaceutical and agrochemical sectors. Aspergillus wentii CBS 141173 has gathered research interest due to its ability to produce high-value norditerpenoid compounds, including anticancer molecules. In this study, we aimed to expand the genomic information available for A. wentii to facilitate the identification of terpenoid biosynthetic genes that may be involved in the production of bioactive molecules.
Long-read genome sequencing of Aspergillus wentii CBS 141173 was conducted using Oxford Nanopore Technologies (ONT) MinION MK1C. In addition, paired-end stranded RNA-seq data from two time points, 7 days and 30 days, was used for functional annotation of the assembled genome. Overall, we assembled a genome of approximately 31.2 Mb and identified 66 biosynthetic gene clusters from the annotated genome. Metabolic extracts of A. wentii were analysed and the production of the bioactive terpenoid asperolide A was confirmed. We further mined the assembled and annotated genome for BGCs involved in terpenoid pathways using a combination of antiSMASH and local BlastP and identified 16 terpene synthases. Phylogenetic analysis was conducted and allowed us to establish relationships with other characterised terpene synthases. We identified two terpene clusters potentially involved in pimarane-like diterpenoid biosynthesis. Finally, the analysis of the 16 terpene synthases in our 7-day and 30-day transcriptomic data suggested that only four of them were constitutively expressed under laboratory conditions.
These results provide a scaffold for the future exploration of terpenoid biosynthetic pathways for bioactive molecules in A. wentii. The terpenoid clusters identified in this study are candidates for heterologous gene expression and/or gene disruption experiments. The description and availability of the long-read genome assembly of A. wentii CBS 141173 further provides the basis for downstream genome analysis and biotechnological exploitation of this species.
真菌是具有应用价值的次生代谢产物的优秀生产者，在制药和农用化学品领域都有涉及。由于其生产高价值的北二萜类化合物（包括抗癌分子）的能力，曲霉属去甲二萜类化合物 CBS 141173 引起了研究兴趣。在这项研究中，我们旨在扩展 A. wentii 的基因组信息，以促进鉴定可能参与生物活性分子生产的萜类生物合成基因。
使用牛津纳米孔技术（ONT）MinION MK1C 对曲霉属去甲二萜 CBS 141173 进行了长读长基因组测序。此外，还使用两个时间点（7 天和 30 天）的配对末端 RNA 测序数据对组装基因组进行了功能注释。总体上，我们组装了一个约 31.2 Mb 的基因组，并从注释的基因组中鉴定了 66 个生物合成基因簇。分析了曲霉属去甲二萜的代谢物提取物，并确认了生物活性萜烯 Asperolide A 的产生。我们进一步使用 antiSMASH 和局部 BlastP 组合对组装和注释的基因组进行挖掘，以鉴定参与萜烯途径的 BGCs，并鉴定了 16 个萜烯合酶。进行了系统发育分析，使我们能够与其他特征化的萜烯合酶建立关系。我们鉴定了两个可能参与 Pimarane 类二萜生物合成的萜烯簇。最后，对我们的 7 天和 30 天转录组数据中的 16 个萜烯合酶的分析表明，在实验室条件下只有其中 4 个是组成型表达的。
这些结果为进一步探索曲霉属去甲二萜生物合成途径提供了一个框架，以获得生物活性分子。本研究中鉴定的萜烯簇是异源基因表达和/或基因敲除实验的候选物。曲霉属去甲二萜 CBS 141173 的长读长基因组组装的描述和可用性进一步为该物种的下游基因组分析和生物技术利用提供了基础。
