Institut für Verfahrenstechnik, Umwelttechnik und Techn. Biowissenschaften
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Unraveling the linkage of morphology and physiology in filamentous fungi by combining PAT tools with automated high–throughput microscopy

 

Wir bitten um Verständnis, dass die Informationen über dieses Projekt derzeit nur in englischer Sprache verfügbar sind.

 

Project Content:

Today, the vast majority of industrial production processes employing filamentous fungi are based on hyper producing strains generated by classical random mutation strain improvement techniques. While recent years have seen the development of target-oriented techniques such as genetic engineering, production strains are still selected by empirical trial-and-error screening procedures inevitably limiting the design space of the production process to suboptimal parameter settings. As a result, complex fungal growth morphology - the unique and highly relevant feature of filamentous fungi - is neglected due to the lack of tools enabling reliable, quantitative analysis.

In order to fulfill the pressing need for a quantitative approach enabling both, physiological and morphological behavior; this work aims at establishing a science-based approach for process development in filamentous fungi meeting QbD (Quality by Design) and PAT (Process Analytical Technology) guidelines linking process dynamics and morphological analysis.

Using P.chrysogenum as a model organism, the adaptation of different industrial candidate strains to altering cultivation conditions in transient cultures is investigated. Dynamic experiments are designed using DoE (Design of Experiment) strategies and carried out in fully instrumented bioreactors coupled to a state-of-the-art process management system allowing online process control and evaluation. Offline data is gathered by high performance liquid and gas chromatography; identification of unknown compounds is enabled by coupling the respective high-resolution method to mass spectrometry. Analysis of fungal morphology is performed using an utterly automated microscopic imaging technique, while morphological image analysis, data integration and statistical data evaluation are facilitated by our newly developed toolbox suitable for general applicability based on the programming language Matlab.

This work presents a methodology allowing accurate quantification of the linkage between filamentous growth morphology and metabolic regulation. By coupling quantitative morphological and metabolic data, modeling and control strategies for complex cellular regulation processes linked to specific morphological structures can be developed.

Our approach combines process technology, growth morphology, metabolism and productivity and delivers a robust, comprehensive model allowing for an optimized design of cultivation process parameters. It will thus be a valuable tool in process development enabling quantification, and - on the other hand - control of production for the wide range of value-added products from filamentous fungi.

 

Industrial Partner:

Biopharmaceutical Producer

 

Scientific Output

 

Kontakt:

Univ.Prof. Dipl.-Ing. Dr.techn. Christoph Herwig

Mitarbeiter:

Dipl.-Ing. Dr.techn. Andreas Posch