Optimisation of the glycerol-to-ornithine molar ratio in the feed medium for the continuous production of clavulanic acid by Streptomyces clavuligerus

Amino acids are well metabolized by Streptomyces clavuligerus during the production of clavulanic acid using glycerol as main carbon and energy source. However, only a few amino acids such as arginine and ornithine are favorable for CA biosynthesis. The aim of this work was to optimize the glycerol:ornithine molar ratio in the feed medium containing only these compounds to maximize CA production in continuous cultivation. A minimum number of experiments were performed by means of a simple two-level full-factorial central composite design to investigate the combined effect of glycerol and ornithine feeding on the CA concentration during the intermittent and continuous process in shake-flasks. Statistical analysis of the experimental data using the response surface methodology showed that a glycerol-to-ornithine molar ratio of approximately 40:1 in the feed medium resulted in the highest CA concentration when fermentation was stopped. Under these optimized conditions, in bench-scale fermentor runs, the CA concentration reached more than double the concentration obtained in shake-flasks runs.