Formal- and high-structured kinetic process modelling and footprint area analysis of binary imaged cells: Tools to understand and optimize multistage-continuous PHA biosynthesis

Abstract

Abstract Competitive polyhydroxyalkanoate (PHAs) production requires progress in microbial strain performance, feedstock selection, downstream processing, and more importantly according to the process design with process kinetics of the microbial growth phase and the phase of product formation. The multistage continuous production in a bioreactor cascade was described for the first time in a continuously operated, flexible five-stage bioreactor cascade that mimics the characteristics involved in the engineering process of tubular plug flow reactors. This process was developed and used for Cupriavidus necator-mediated PHA production at high volumetric and specific PHA productivity (up to 2.31 g/(Lh) and 0.105 g/(gh), respectively). Based on the experimental data, formal kinetic and high structured kinetic models were established, accompanied by footprint area analysis of binary imaged cells. As a result of the study, there has been an enhanced understanding of the long-term continuous PHA production under balanced, transient, and nutrient-deficient conditions that was achieved on both the micro and the macro kinetic level. It can also be concluded that there were novel insights into the complex metabolic occurrences that developed during the multistage- continuous production of PHA as a secondary metabolite. This development was essential in paving the way for further process improvement. At the same time, a new method of specific growth rate and specific production rate based on footprint area analysis was established by using the electron microscope.