Analysis and effect of conventional flasks in shaking culture of Escherichia coli
Masato Takahashi, Hideki Aoyagi
Abstract
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Masato Takahashi, Hideki Aoyagi
Abstract
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Abstract 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 kLa, an index for evaluating the performance of aerobic culture incubators, and kG, 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 kG than the Erlenmeyer and Sakaguchi flasks, and yielded kLa 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 kLa 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 kLa and kG may contribute to a wide range of fields employing microorganism culture.
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Abstract 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 kLa, an index for evaluating the performance of aerobic culture incubators, and kG, 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 kG than the Erlenmeyer and Sakaguchi flasks, and yielded kLa 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 kLa 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 kLa and kG may contribute to a wide range of fields employing microorganism culture.
Key concepts: Erlenmeyer flask, Laboratory flask, Chromatography, Bioreactor, Microbiology, Chemistry, Biology, Botany