Han Hongbee, Liu Rundong, Woo Jung-Jae, Hur Jae-Seoun, Kim Wonyong
Korean Lichen Research Institute, Sunchon National University, Suncheon, South Korea.
Department of Applied Biology, College of Agriculture and Life Sciences, Chonnam National University, Gwangju, South Korea.
Mycobiology. 2025 Feb 13;53(2):225-235. doi: 10.1080/12298093.2025.2460292. eCollection 2025.
The plant pathogenic fungus produces solanapyrones, which are polyketide-derived secondary metabolites, during its saprobic growth on straws. Previously, we utilized as a heterologous host to produce a lichen-derived polyketide with anticancer activities. This study aimed to establish as a sustainable biorefinery for production of biologically active compounds through two objectives: optimizing culture conditions on agricultural waste substrates for polyketide production and generating a clean host using the Cre-loxP system for reusable antibiotic resistance markers. We found that soy hull is the most effective substrate among lignocellulosic materials. Neither light nor the addition of extra divalent cations were required for solanapyrone production in . Production of solanapyrones peaked 18 days after culture on soy hull pellets. To generate a clean host, we deleted the gene, involved in melanin biosynthesis, and the gene, responsible for solanapyrone production. Antibiotic resistance markers used in genetic transformation were recycled by establishing a Cre-loxP system in . In this system, Cre recombinase was expressed under the control of a promoter inducible during sporulation to mitigate the cytotoxicity of Cre. Here, we set the groundwork for developing as an environmentally-friendly biorefinery by generating a clean host with a Cre-loxP marker recycling system and optimizing growth conditions with soy hull pellets.
这种植物病原真菌在秸秆上腐生生长时会产生茄病镰刀菌烯酮,它们是聚酮化合物衍生的次生代谢产物。此前,我们利用[具体名称未给出]作为异源宿主来生产一种具有抗癌活性的地衣衍生聚酮化合物。本研究旨在通过两个目标将[具体名称未给出]建立为生产生物活性化合物的可持续生物精炼厂:优化农业废弃物底物上的聚酮化合物生产培养条件,以及使用Cre-loxP系统生成一个干净的宿主以实现可重复使用的抗生素抗性标记。我们发现大豆皮是木质纤维素材料中最有效的底物。在[具体名称未给出]中生产茄病镰刀菌烯酮既不需要光照也不需要添加额外的二价阳离子。在大豆皮颗粒上培养18天后,茄病镰刀菌烯酮的产量达到峰值。为了生成一个干净的宿主,我们删除了参与黑色素生物合成的[具体基因未给出]基因和负责茄病镰刀菌烯酮生产的[具体基因未给出]基因。通过在[具体名称未给出]中建立Cre-loxP系统,遗传转化中使用的抗生素抗性标记得以循环利用。在这个系统中,Cre重组酶在孢子形成过程中可诱导的启动子控制下表达,以减轻Cre的细胞毒性。在此,我们通过生成具有Cre-loxP标记循环系统的干净宿主并优化大豆皮颗粒的生长条件,为将[具体名称未给出]发展成为环境友好型生物精炼厂奠定了基础。