Institute of Medicinal Biotechnology, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, 100050, China.
National Key Laboratory of Chinese Medicine Modernization, State Key Laboratory of Component-Based Chinese Medicine, Institute of Traditional Chinese Medicine, Tianjin University of Traditional Chinese Medicine, Tianjin, 301617, China.
Appl Microbiol Biotechnol. 2024 Sep 12;108(1):462. doi: 10.1007/s00253-024-13282-4.
Bioinformatic analysis revealed that the genomes of ubiquitous Penicillium spp. might carry dozens of biosynthetic gene clusters (BGCs), yet many clusters have remained uncharacterized. In this study, a detailed investigation of co-culture fermentation including the basidiomycete Armillaria mellea CPCC 400891 and the P. brasilianum CGMCC 3.4402 enabled the isolation of five new compounds including two bisabolene-type sesquiterpenes (arpenibisabolanes A and B), two carotane-type sesquiterpenes (arpenicarotanes A and B), and one polyketide (arpenichorismite A) along with seven known compounds. The assignments of their structures were deduced by the extensive analyses of detailed spectroscopic data, electronic circular dichroism spectra, together with delimitation of the biogenesis. Most new compounds were not detected in monocultures under the same fermentation conditions. Arpenibisabolane A represents the first example of a 6/5-fused bicyclic bisabolene. The bioassay of these five new compounds exhibited no cytotoxic activities in vitro against three human cancer cell lines (A549, MCF-7, and HepG2). Moreover, sequence alignments and bioinformatic analysis to other metabolic pathways, two BGCs including Pb-bis and Pb-car, responsible for generating sesquiterpenoids from co-culture were identified, respectively. Furthermore, based on the chemical structures and deduced gene functions of the two clusters, a hypothetic metabolic pathway for biosynthesizing induced sesquiterpenoids was proposed. These results demonstrated that the co-culture approach would facilitate bioprospecting for new metabolites even from the well-studied microbes. Our findings would provide opportunities for further understanding of the biosynthesis of intriguing sesquiterpenoids via metabolic engineering strategies. KEY POINTS: • Penicillium and Armillaria co-culture facilitates the production of diverse secondary metabolites • Arpenibisabolane A represents the first example of 6/5-fused bicyclic bisabolenes • A hypothetic metabolic pathway for biosynthesizing induced sesquiterpenoids was proposed.
生物信息学分析表明，普遍存在的青霉属（Penicillium spp.）的基因组可能携带数十个生物合成基因簇（BGCs），但许多簇仍未被描述。在这项研究中，对共生培养发酵的详细研究包括担子菌蜜环菌（Armillaria mellea CPCC 400891）和青霉（P. brasilianum CGMCC 3.4402），使我们能够分离出五种新化合物，包括两种倍半萜（arpenibisabolanes A 和 B）、两种胡萝卜烷型倍半萜（arpenicarotanes A 和 B）和一种聚酮化合物（arpenichorismite A）以及七种已知化合物。通过详细的光谱数据分析、电子圆二色性光谱以及生物发生的限定，推断出它们的结构。在相同的发酵条件下，在单培养中没有检测到大多数新化合物。Arpenibisabolane A 代表了第一个 6/5 稠合双环倍半萜的例子。这些五种新化合物的生物测定显示，它们在体外对三种人类癌细胞系（A549、MCF-7 和 HepG2）没有细胞毒性。此外，通过序列比对和生物信息学分析，确定了负责从共生培养物中产生倍半萜的两个 BGCs，分别是 Pb-bis 和 Pb-car。此外，基于两个簇的化学结构和推断的基因功能，提出了一个假设的生物合成诱导倍半萜的代谢途径。这些结果表明，共生培养方法有助于从研究良好的微生物中发现新的代谢产物。我们的发现为通过代谢工程策略进一步了解有趣的倍半萜生物合成提供了机会。
青霉属和蜜环菌共生培养促进了多种次生代谢产物的产生。
Arpenibisabolane A 代表了第一个 6/5 稠合双环倍半萜的例子。
提出了一个假设的生物合成诱导倍半萜的代谢途径。
