Filamentous co-cultures: from screening to bioprocess development (Co-Pilot)

丝状共培养物：从筛选到生物工艺开发（Co-Pilot）

• 批准号: 427899901
• 负责人: Professorin Dr. Miriam Agler-Rosenbaum
• 金额: --
• 依托单位: Leibniz-Institut für Naturstoff-Forschung und Infektionsbiologie e. V. Hans-Knöll-Institut
• 依托单位国家: 德国
• 项目类别: Priority Programmes
• 资助国家: 德国
• 项目状态: 未结题
• 来源: https://gepris.dfg.de/gepris/projekt/427899901?language=en
• 关键词: Filamentous; co; cultures; screening; bioprocess

The co-cultivation of cellulolytic filamentous fungi and filamentous soilbacteria on cellulosic substrates represents a promising approach forthe discovery and production of natural products. Due to thecomplexity, filamentous co-cultures are poorly studied andcharacterized. In the proposed project, the cellulose degraderTrichoderma reesei is partnered with various Streptomyces strains tosystematically unlock the production of novel natural products in thelatter genus. The work is based on our experience with a model cocultureof T. reesei and Streptomyces coelicolor, which is well knownto produce pigments and antibiotics. For this co-culture, weestablished a fluorescence-based method to follow populationdynamics and non-invasive methods for analysis of metabolism andpigment formation. To enable the screening of population dynamics ofmany different co-cultures using non-tagged Streptomyces strains, amethod based on autofluorescence profiling in combination withfluorescently-tagged T. reesei will be established. By meta-analysis ofaxenic and co-cultures of different Streptomyces strains, correlationsbetween the axenic performance indicators (e.g. growth rate) and coculturebehavior can be established. Thereby, we especially expectgreat performance differences depending on morphology. Tosystematically analyze the influence of morphology, co-cultures of adispersed growing knockout strain of our model bacterium S.coelicolor will be compared to our established co-culture of the pelletgrowing wildtype strain of S. coelicolor. Thereby, population stability,product formation, morphology, and rheology will be monitored andcorrelated to the different morphology:performance results of theStreptomyces screening. This will allow to estimate to what extentStreptomyces co-culture behavior can be generalized and predicted,which will aid in future design of filamentous co-cultures.Subsequently, a systematic up-scaling from microtiter plate to stirredfermenter will be performed for the most promising co-cultivationsdiscovered during the screening. Due to the morphologicalcomplexity, the co-culture morphology and performance is assumed to be sensitive towards changes in fluid dynamics. Therefore, scaleupwill be compared based on maximum local and average volumetricpower input by assessment of different impellers under closeinvestigation of morphology and rheology. All measurementtechniques already applied in small scale will be used to study howthe biosynthetic performance of filamentous co-cultures changeacross scales. With this information, we will be able to steer theinteractions of co-culture partners and enable a straightforwardmorphology-based scale-up to the pilot scale.

纤维素分解丝状真菌和土壤丝状细菌在纤维素基质上的共培养代表了发现和生产天然产物的一种有前途的方法。由于复杂性，丝状共培养物的研究和表征很少。在拟议的项目中，纤维素降解剂里氏木霉与各种链霉菌菌株合作，系统地解锁后一种属的新型天然产物的生产。这项工作是基于我们的经验与模型cocultureof T。reesei和天蓝色链霉菌，这是众所周知的产生色素和抗生素。对于这种共培养，我们建立了一种基于荧光的方法来跟踪种群动态和非侵入性方法来分析代谢和色素形成。为了能够筛选使用非标记链霉菌菌株的许多不同共培养物的群体动力学，基于自发荧光谱结合荧光标记的T.将建立瑞泽。通过对不同链霉菌菌株的无菌和共培养物的荟萃分析，可以建立无菌性能指标（如生长速率）与共培养行为之间的相关性。因此，我们特别期待巨大的性能差异取决于形态。为了系统地分析形态学的影响，将我们的模式细菌天蓝色链霉菌的分散生长敲除菌株的共培养物与我们建立的生长颗粒的野生型菌株的共培养物进行比较。天蓝色。因此，群体稳定性、产物形成、形态和流变学将被监测并与链霉菌筛选的不同形态：性能结果相关联。这将允许估计在何种程度上链霉菌共培养行为可以概括和预测，这将有助于未来的丝状共培养物的设计。随后，将进行系统的放大，从微量滴定板搅拌发酵罐筛选过程中发现的最有前途的共培养物。由于形态的复杂性，共培养的形态和性能被认为是对流体动力学的变化敏感。因此，将根据最大局部和平均体积功率输入，通过在形态和流变学的密切调查下评估不同的叶轮来比较放大。所有已经在小规模应用的测量技术将被用来研究丝状共培养物的生物合成性能如何在不同规模上变化。有了这些信息，我们将能够引导共培养伙伴的相互作用，并使基于形态学的直接规模扩大到中试规模。