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

doi:10.1039/d0ra02803g

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 journal › Article › peer-review

3 Citations (Scopus)

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 language	English
Pages (from-to)	16679-16686
Number of pages	8
Journal	RSC Advances
Volume	10
Issue number	28
DOIs	https://doi.org/10.1039/d0ra02803g
Publication status	Published - Apr 28 2020

All Science Journal Classification (ASJC) codes

Chemistry(all)
Chemical Engineering(all)

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10.1039/d0ra02803g

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title = "Spatiotemporal monitoring of intracellular metabolic dynamics by resonance Raman microscopy with isotope labeling",

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.",

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

note = "Funding Information: This work was supported primarily by the ImPACT Program of the CSTI (Cabinet Office, Government of Japan) and partly by the JSPS Core-to-Core Program and White Rock Foundation. The authors thank Prof. Tomohisa Hasunuma at Kobe University for discussion. Publisher Copyright: {\textcopyright} The Royal Society of Chemistry 2020.",

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AU - Yonamine, Yusuke

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AU - Fujiwara, Tomomi

AU - Miura, Yoshiko

AU - Goda, Keisuke

AU - Hoshino, Yu

N1 - Funding Information: This work was supported primarily by the ImPACT Program of the CSTI (Cabinet Office, Government of Japan) and partly by the JSPS Core-to-Core Program and White Rock Foundation. The authors thank Prof. Tomohisa Hasunuma at Kobe University for discussion. Publisher Copyright: © The Royal Society of Chemistry 2020.

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N2 - 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.

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

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