Analysis and effect of conventional flasks in shaking culture of Escherichia coli

The circulation direct monitoring and sampling system (CDMSS) is used as a monitoring device for CO₂ and O₂ concentrations of bypass type in shake-culture flask. The CDMSS could measure k_La, an index for evaluating the performance of aerobic culture incubators, and k_G, an indicator of the degree of CO₂ ventilation in the flask gas phase. We observed that cylindrical flasks provided a different culture environment, yielded a much higher k_G than the Erlenmeyer and Sakaguchi flasks, and yielded k_La equivalent to that by Erlenmeyer flask by setting the ring-type baffle appropriately. Baffled cylindrical flask used for Escherichia coli K12 IFO3301 shake culture maintained lower CO₂ concentrations in the headspace than conventional flasks; therefore, CO₂ accumulation in the culture broth could be suppressed. Cell growth in baffled cylindrical flask (with k_La equivalent to that of the Erlenmeyer flask) was about 1.3 and 1.4 times that in the Erlenmeyer and Sakaguchi flasks, respectively. This study focused on the batch culture at the flask scale and designed the headspace environment with low CO₂ accumulation. Therefore, we conclude that redesign of flasks based on k_La and k_G may contribute to a wide range of fields employing microorganism culture.