Takekana Musashi, Yoshida Takanobu, Yoshida Erika, Ono Sumika, Horie Shinnosuke, Vavricka Christopher J, Hiratani Moe, Tsuge Kenji, Ishii Jun, Hayakawa Yoshihiro, Kondo Akihiko, Hasunuma Tomohisa
Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan.
Graduate School of Science, Technology and Innovation, Kobe University, Kobe, Japan; Research Institute for Bioscience Products & Fine Chemicals. Ajinomoto Co., Inc. Kanagawa, Japan.
J Chromatogr B Analyt Technol Biomed Life Sci. 2023 Jan 15;1215:123588. doi: 10.1016/j.jchromb.2022.123588. Epub 2022 Dec 28.
Conventional analysis of microbial bioproducers requires the extraction of metabolites from liquid cultures, where the culturing steps are time consuming and greatly limit throughput. To break through this barrier, the current study aims to directly evaluate microbial bioproduction colonies by way of supercritical fluid extraction-supercritical fluid chromatography-triple quadrupole mass spectrometry (SFE-SFC-MS/MS). The online SFE-SFC-MS/MS system offers great potential for high-throughput analysis due to automated metabolite extraction without any need for pretreatment. This is the first report of SFE-SFC-MS/MS as a method for direct colony screening, as demonstrated in the high-throughput screening of (-)-limonene bioproducers. Compared with conventional analysis, the SFE-SFC-MS/MS system enables faster and more convenient screening of highly productive strains.
对微生物生物生产者的传统分析需要从液体培养物中提取代谢物，其中培养步骤耗时且极大地限制了通量。为突破这一障碍，当前研究旨在通过超临界流体萃取-超临界流体色谱-三重四极杆质谱联用技术（SFE-SFC-MS/MS）直接评估微生物生物生产菌落。在线SFE-SFC-MS/MS系统由于无需任何预处理即可自动进行代谢物提取，因此具有高通量分析的巨大潜力。这是首次报道将SFE-SFC-MS/MS作为直接菌落筛选方法，如在(-)-柠檬烯生物生产者的高通量筛选中所展示的那样。与传统分析相比，SFE-SFC-MS/MS系统能够更快、更方便地筛选高产菌株。
