Single cell analysis and quantitative physiology of industrially relevant microorganisms

Viac o knihe

The analysis of single cell behavior is a crucial area of research in industrial biotechnology, allowing for the study of microorganisms without relying on average population values. Lab-on-a-chip (LOC) devices facilitate controlled single cell cultivations, enabling detailed analysis of cellular physiology and extracellular environments. This thesis presents the development of a novel microfluidic LOC device for single cell cultivation and analytical methods to investigate cellular physiology at an individual level. A new manufacturing process created a versatile, chemically inert Envirostat for contactless cultivation, offering exceptional operational stability and defined surface chemistry, which opens up new analytical opportunities for single cell secretome analyses. Various analytical technologies were assessed for their sensitivity to amino acids, key biotechnological products of the bacterium Corynebacterium glutamicum. Methodologies utilizing optical fluorescence readouts were also employed. The study identified individual medium components that either hindered or enhanced L-lysine production by C. glutamicum. Additionally, a regulatory effect of glucose availability on substrate uptake rate was discovered, laying the groundwork for further research into the metabolic control mechanisms of C. glutamicum. Overall, this work developed and applied innovative technologies and methods to explore key parameters of cellular