Control of mono-species and multi-species pellet heterogeneity and their implications on product formation in the cell factory Aspergillus niger

单物种和多物种颗粒异质性的控制及其对黑曲霉细胞工厂中产物形成的影响

• 批准号: 427889137
• 负责人: Professor Dr.-Ing. Heiko Briesen
• 金额: --
• 依托单位: Fachgebiet Angewandte und Molekulare Mikrobiologie
• 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/427889137?language=en
• 关键词: Control; mono; species; multi; pellet

Filamentous fungi including Aspergillus niger are cell factories in biotechnology for the production of organic acids, proteins and bioactive compounds. However, the evolution of the fungal macroscopic morphologies such as dispersed mycelia and pellets remains an unpredictable multifactorial process, which strongly limits productivities.The overall objective of the project is to gain mechanistic insights into the heterogeneous macromorphological characteristics of A. niger in stirred tank bioreactor (STR) and rocking-motion bioreactor (RMB) cultivations - within individual and among macromorphological populations. Although heterogeneities within a population and within bioreactor compartments are known for a long time, the genetic basis of filamentous heterogeneity as well as options for rational process design have neither been studied so far in a systematic manner nor are cell-cell and cell-bioreactor interactions comprehensively understood for filamentous fungi. To comprehensively understand the evolution of mono-species (A. niger) and multi-species (A. niger and S. coelicolor) pellet morphologies in lab-scale and large scale bioreactors, their impact on the established product portfolio of A. niger (primary metabolite citric acid, enzyme glucoamylase, secondary metabolite enniatin B) and the induction of new products (secondary metabolites) due to the presence of the filamentous bacterium S. coelicolor, we will test different STR and RMB designs and inoculation schemes. The impact of these factors on growth, physiology, morphology and product formation will be studied using on line and off line microscopic tools, viability analyses, transcriptomics, metabolomics, and X-ray µ-computed tomography, which characterize the 3-dimensional inner structure of pellets. The ultimate goal is in understanding both genetic and process engineering mechanisms behind the development of population heterogeneities that balance hyphal growth with product formation. Understanding the link between gene regulatory and metabolic pathways with changing process environments will ultimately identify gene switches in A. niger whose targeted manipulation will rationally improve productivity of this cell factory during industrial fermentation – not only with respect to product formation, but also with respect to improved macromorphologies. Understanding the effect of process design on the biological system will in turn generate leads for optimized process control. We further propose the establishment of a multiscale modelling platform which couples the evolution of macromorphological heterogeneities of filamentous microorganisms with oxygen supply and product formation. Such model framework will eventually support model-based process design for improved bioprocesses. Hence, rational genetic and process engineering to predict and control macroscopic morphologies of A. niger in axenic or mixed bioprocesses will become possible.

丝状真菌包括尼日尔曲霉是生物技术中用于生产有机酸、蛋白质和生物活性化合物的细胞工厂。然而，真菌宏观形态的进化（如分散的菌丝体和小球）仍然是一个不可预测的多因素过程，这严重限制了生产力。黑曲霉在搅拌罐生物反应器（STR）和摇摆运动生物反应器（RMB）培养-在个人和之间的macromorphological群体。虽然种群内和生物反应器隔室内的异质性是已知的很长一段时间，丝状异质性的遗传基础，以及合理的工艺设计的选择，迄今为止既没有研究过系统的方式，也没有全面了解丝状真菌的细胞-细胞和细胞-生物反应器的相互作用。为了全面了解单物种的进化（A。尼日尔）和多种（A.尼日尔和S. coelicolor）颗粒形态，它们对A.尼日尔（初级代谢产物柠檬酸、葡糖淀粉酶、次级代谢产物恩镰孢菌素B）和由于丝状细菌S. coelicolor，我们将测试不同的STR和RMB设计和接种方案。将使用在线和离线显微镜工具、活力分析、转录组学、代谢组学和X射线微计算机断层扫描（表征颗粒的三维内部结构）研究这些因素对生长、生理学、形态学和产品形成的影响。最终目标是了解平衡菌丝生长与产物形成的群体异质性发展背后的遗传和工艺工程机制。了解基因调控和代谢途径与不断变化的工艺环境之间的联系，将最终确定A。尼日尔，其靶向操作将合理地提高该细胞工厂在工业发酵期间的生产率-不仅在产物形成方面，而且在改进的宏观形态方面。了解工艺设计对生物系统的影响将反过来产生优化工艺控制的线索。我们进一步建议建立一个多尺度的建模平台，该平台将丝状微生物的宏观形态异质性的进化与氧气供应和产物形成相结合。这种模型框架将最终支持基于模型的工艺设计，以改进生物过程。因此，合理的遗传工程和过程工程来预测和控制宏观形态。在纯或混合生物过程中的黑尼日尔将成为可能。