Müller Henri, Deffur Charlotte, Schmideder Stefan, Barthel Lars, Friedrich Tiaan, Mirlach Lukas, Hammel Jörg U, Meyer Vera, Briesen Heiko
School of Life Sciences Weihenstephan, Chair of Process Systems Engineering, Technical University of Munich, Freising, Germany.
Chair of Applied and Molecular Microbiology, Institute of Biotechnology, Technische Universität Berlin, Berlin, Germany.
Biotechnol Bioeng. 2023 Nov;120(11):3244-3260. doi: 10.1002/bit.28506. Epub 2023 Jul 21.
Filamentous fungi produce a wide range of relevant biotechnological compounds. The close relationship between fungal morphology and productivity has led to a variety of analytical methods to quantify their macromorphology. Nevertheless, only a µ-computed tomography (µ-CT) based method allows a detailed analysis of the 3D micromorphology of fungal pellets. However, the low sample throughput of a laboratory µ-CT limits the tracking of the micromorphological evolution of a statistically representative number of submerged cultivated fungal pellets over time. To meet this challenge, we applied synchrotron radiation-based X-ray microtomography at the Deutsches Elektronen-Synchrotron [German Electron Synchrotron Research Center], resulting in 19,940 3D analyzed individual fungal pellets that were obtained from 26 sampling points during a 48 h Aspergillus niger submerged batch cultivation. For each of the pellets, we were able to determine micromorphological properties such as number and density of spores, tips, branching points, and hyphae. The computed data allowed us to monitor the growth of submerged cultivated fungal pellets in highly resolved 3D for the first time. The generated morphological database from synchrotron measurements can be used to understand, describe, and model the growth of filamentous fungal cultivations.
丝状真菌能产生多种相关的生物技术化合物。真菌形态与生产力之间的密切关系催生了多种量化其宏观形态的分析方法。然而,只有基于µ-计算机断层扫描(µ-CT)的方法能够对真菌菌球的三维微观形态进行详细分析。然而,实验室µ-CT的低样品通量限制了对大量具有统计学代表性的深层培养真菌菌球随时间的微观形态演变的追踪。为应对这一挑战,我们在德国电子同步加速器研究中心应用了基于同步辐射的X射线显微断层扫描技术,在黑曲霉48小时深层分批培养过程中的26个采样点获取了19940个经过三维分析的单个真菌菌球。对于每个菌球,我们能够确定其微观形态特征,如孢子、尖端、分支点和菌丝的数量及密度。计算得到的数据使我们首次能够在高分辨率三维空间中监测深层培养真菌菌球的生长。同步辐射测量生成的形态学数据库可用于理解、描述和模拟丝状真菌培养物的生长。