Optimization of fermentation medium composition in substrate-controlled continuous stirred tank reactors

A 'carbon source controlled shift technique' was developed for fermentation medium optimization in continuous culture with the objective to maximize growth rate and growth-linked product formation of a biological system. An automatic culture medium preparation system was operated together with 2 parallel stirred tank reactors and a HPLC system for on-line analysis of the carbon source concentration in the reactors. A genetic algorithm was applied for experimental design. The concentrations of 7 medium components (mineral salts and vitamins) were optimized automatically within 40 continuous experiments to result in a maximum growth rate of the methylotrophic yeast Candida boidinii and growth-linked production of the formate dehydrogenase enzyme (FDH). The specific growth rate of Candida boidinii and the specific activity of the FDH enzyme at a set-point of 420 mM methanol in the reactor were, thus, improved by 19% to 0.16 h^-1 and 26% to 164 U g^-1, respectively, compared to the previously used medium, which has already been previously optimized in shake flask experiments. The results of the continuous medium optimization were evaluated with a full second order seven-dimensional polynomial model (regression coefficient 96.8%).