Aspochalasin D（AD）属于聚酮氨基酸混合天然产物，具有抗癌、抗菌和防污染生物活性。然而，低产量限制了其进一步应用。在这项研究中，AD从Aspergillus flavipes 3.17641分离并鉴定。接下来，在进行单因素实验和响应面方法优化培养条件的同时，采用代谢工程学增加AD产量。结果表明，删去旁路基因aspoA并过表达特定通路调节因子aspoG显著提高AD产量。在优化条件下，其产量达到812.1毫克/升，增加了18.5倍。因此，本研究不仅提供了一般方法来提高其他真菌类似天然产物的产量，而且使AD在农业和医药领域的生物学功能进一步得以开发。关键点：•优化培养条件提高了Aspochalasin D（AD） 的产量。•删去旁路基因aspoA增加了AD的产量。•过表达通路调节因子aspoG进一步提高了AD的产量。 © 2023年作者，独家许可给Springer-Verlag GmbH Germany，Springer Nature的一部分。

Aspochalasin D (AD) belongs to the polyketide-amino acid hybrid natural products with anti-cancer, anti-bacterial, and anti-fouling bioactivities. However, the low production limits its further application. In this study, AD was separated and identified from Aspergillus flavipes 3.17641. Next, besides the optimization of culture conditions using a single-factor experiment and response surface methodology, metabolic engineering was employed to increase the AD production. It shows that the deletion of the shunt gene aspoA and overexpression of the pathway-specific regulator aspoG significantly improve the AD production. Its production reached to 812.1 mg/L under the optimized conditions, with 18.5-fold increase. Therefore, this study not only provides a general method for improving the production of similar natural products in other fungi, but also enables the further biological function development of AD in agriculture and pharmaceutical. KEY POINTS: • The Aspochalasin D (AD) production was improved by optimizing culture conditions. • The deletion of the shunt gene aspoA increased the AD production. • Overexpression of the pathway regulator aspoG further improved the AD production.© 2023. The Author(s), under exclusive licence to Springer-Verlag GmbH Germany, part of Springer Nature.