Section: New Results

Growth control in bacteria and biotechnological applications

The ability to experimentally control the growth rate is crucial for studying bacterial physiology. It is also of central importance for applications in biotechnology, where often the goal is to limit or even arrest growth. Growth-arrested cells with a functional metabolism open the possibility to channel resources into the production of a desired metabolite, instead of wasting nutrients on biomass production. In recent years we obtained a foundation result for growth control in bacteria [6], in that we engineered an E. coli strain where the transcription of a key component of the gene expression machinery, RNA polymerase, is under the control of an inducible promoter. By changing the inducer concentration in the medium, we can adjust the RNA polymerase concentration and thereby switch bacterial growth between zero and the maximal growth rate supported by the medium. The publication also presented a biotechnological application of the synthetic growth switch in which both the wild-type E. coli strain and our modified strain were endowed with the capacity to produce glycerol when growing on glucose. Cells in which growth has been switched off continue to be metabolically active and harness the energy gain to produce glycerol at a twofold higher yield than in cells with natural control of RNA polymerase expression.

The experimental work underlying the growth switch has been continued in several directions in the context of the Maximic project by Célia Boyat. Moreover, in collaboration with colleagues from the BIOCORE project-team, we have formulated the maximization of metabolite production by means of the growth switch as a resource reallocation problem that can be analyzed by means of the self-replicator models of bacterial growth in combination with methods from optimal control theory. In a paper published in the Journal of Mathematical Biology this year [24], we study various optimal control problems by means of a combination of analytical and computational techniques. We show that the optimal solutions for biomass maximization and product maximization are very similar in the case of unlimited nutrient supply, but diverge when nutrients are limited. Moreover, external growth control overrides natural feedback growth control and leads to an optimal scheme consisting of a first phase of growth maximization followed by a second phase of product maximization. This two-phase scheme agrees with strategies that have been proposed in metabolic engineering. More generally, this work shows the potential of optimal control theory for better understanding and improving biotechnological production processes. Extensions concerning the effect on growth and bioproduction of the (biological or technological) costs associated with discontinuous control strategies, and of the time allotted to optimal substrate utilization, were presented at the European Control Conference (ECC 2019) in Naples this year and published in the proceedings [25].