Analysis of effects of flask gas phase on microorganisms during shaking culture and proposal of new shaking culture method

2021 Fiscal Year Final Research Report

• Project/Area Number: 20K15097
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 27040:Biofunction and bioprocess engineering-related
• Research Institution: University of Tsukuba
• Principal Investigator: Takahashi Masato 筑波大学, 生命環境系, 研究員 (60826965)
• Project Period (FY): 2020-04-01 – 2022-03-31
• Keywords: 液内振盪培養 / 振盪フラスコ培養法 / モニタリングデバイス / 二酸化炭素換気能 / 酸素供給能 / 微生物 / ガス制御

Outline of Final Research Achievements

This study was challenged to "clarify the conventional culture environment" and "create the unexplored culture environment". In order to understand the microbial culture environment that can be set by the conventional shake-flask culture conditions, the oxygen (O2) supply capacity and carbon dioxide (CO2) ventilation capacity of various flask conditions, focusing on the shape of the flasks and the type of culture-stoppers were quantified. As a result, it was cleared that CO2 ventilation capacity was much smaller than O2 supply capacity during shaking culture used by Erlenmeyer flasks and Sakaguchi flasks. Therefore, novel device was developed for enhancing ventilation to shake-flask systems. The small device does not require additional electrical power.

Free Research Field

培養工学

Academic Significance and Societal Importance of the Research Achievements

振盪フラスコ培養法は、微生物を用いた生産技術の開発や改善を図るうえで、作業しやすいスケールで低コストかつ多検体を検討できる重要な液内培養技術である。これまで意図せず換気不十分で使用されてきたため、非意図的な二酸化炭素の影響が潜在していたことが明らかとなった。開発した小型の換気デバイス（外部電力が不要で、フラスコを設置する振盪基盤のデッドスペースに外付けできる）を付与することで、液内振盪培養中のフラスコ気相部の培養環境を制御でき、バイオプロセス開発上流の培養条件の検討や、培養を介した微生物種および物質生産のスクリーニングに期待できる。