Spatiotemporal monitoring of intracellular metabolic dynamics by resonance Raman microscopy with isotope labeling

Yusuke Yonamine, Kotaro Hiramatsu, Takuro Ideguchi, Takuro Ito, Tomomi Fujiwara, Yoshiko Miura, Keisuke Goda, Yu Hoshino

Research output: Contribution to journalArticlepeer-review

Abstract

Cellular metabolites are valuable in a diverse range of applications. For example, the unicellular green algaHaematococcus lacustrisproduces as a secondary metabolite the carotenoid pigment astaxanthin (AXT), which is widely used in nutraceutical, cosmetic, and food industries due to its strong antioxidant activity. In order to enhance the productivity ofH. lacustris, spatial and temporal understanding of its metabolic dynamics is essential. Here we show spatiotemporal monitoring of AXT production inH. lacustriscells by resonance Raman microscopy combined with stable isotope labeling. Specifically, we incorporated carbon dioxide (13CO2) labeled with a stable isotope (13C) intoH. lacustriscells through carbon fixation and traced its conversion to13C-AXT using our resonance Raman microscope. We incubatedH. lacustriscells under various conditions by switching, pulsing, and replacing13CO2and12CO2. By measurement of these cells we determined the fixation time of13C-carbon, visualized the intracellular localization of13C- and12C-AXTs, and revealed the dynamic consumption-production equilibrium of the accumulated AXT. This work is a valuable step in the development of effective screening criteria for high AXT-producingH. lacustriscells.

Original languageEnglish
Pages (from-to)16679-16686
Number of pages8
JournalRSC Advances
Volume10
Issue number28
DOIs
Publication statusPublished - Apr 28 2020

All Science Journal Classification (ASJC) codes

  • Chemistry(all)
  • Chemical Engineering(all)

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