Jung Minseo, Lee Jinwon, Park Si Jae, Na Jeong-Geol
Department of Chemical and Biomolecular Engineering Sogang University Mapo-gu Seoul Republic of Korea.
Division of Chemical Engineering and Materials Science Ewha Womans University Seodaemun-gu Seoul Republic of Korea.
Eng Life Sci. 2024 Mar 29;24(7):e2300243. doi: 10.1002/elsc.202300243. eCollection 2024 Jul.
Shake flask cultivation, a cornerstone in bioprocess research encounters limitations in supplying sufficient oxygen and exchanging gases, restricting its accuracy in assessing microbial growth and metabolic activity. In this communication, we introduce an innovative gas supply apparatus that harnesses the rotational motion of a shaking incubator to facilitate continuous air delivery, effectively overcoming these limitations. We measured the mass transfer coefficient (ka) and conducted batch cultures of H36LsGAD using various working volumes to assess its performance. Results demonstrated that the gas supply apparatus significantly outperforms conventional silicone stoppers regarding oxygen delivery, with ka values of 2531.7 h compared to 20.25 h at 230 rpm. Moreover, in batch cultures, the gas supply apparatus enabled substantial improvements in microbial growth, maintaining exponential growth even at larger working volumes. Compared to the existing system, an increase in final cell mass by a factor of 3.4-fold was observed when utilizing 20% of the flask's volume, and a remarkable 9-fold increase was achieved when using 60%. Furthermore, the gas supply apparatus ensured consistent oxygen supply and efficient gas exchange within the flask, overcoming challenges associated with low working volumes. This approach offers a simple yet effective solution to enhance gas transfer in shake flask cultivation, bridging the gap between laboratory-scale experiments and industrial fermenters. Its broad applicability holds promise for advancing research in bioprocess optimization and scale-up endeavors.
摇瓶培养是生物过程研究的基石，但在提供充足氧气和气体交换方面存在局限性，限制了其评估微生物生长和代谢活性的准确性。在本通讯中，我们介绍了一种创新的气体供应装置，该装置利用振荡培养箱的旋转运动促进连续空气输送，有效克服了这些局限性。我们测量了传质系数（ka），并使用不同的工作体积对H36LsGAD进行了分批培养，以评估其性能。结果表明，在氧气输送方面，该气体供应装置明显优于传统的硅胶塞，在230转/分钟时，其ka值为2531.7小时，而传统硅胶塞的ka值为20.25小时。此外，在分批培养中，该气体供应装置显著改善了微生物生长，即使在较大的工作体积下也能保持指数生长。与现有系统相比，当使用20%的摇瓶体积时，最终细胞质量增加了3.4倍，当使用60%的摇瓶体积时，最终细胞质量显著增加了9倍。此外，该气体供应装置确保了摇瓶内氧气的持续供应和高效气体交换，克服了与低工作体积相关的挑战。这种方法为增强摇瓶培养中的气体传递提供了一种简单而有效的解决方案，弥合了实验室规模实验与工业发酵罐之间的差距。其广泛的适用性有望推动生物过程优化和放大研究的进展。
