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Single-step purified R-phycoerythrin transmits cellular imaging functionalities in vitro.

Malairaj Sathuvan1, Ramar Thangam2, Gopal Venkateshbabu3

  • 1Department of Biology & Guangdong Provincial Key Laboratory of Marine Biotechnology, Institute of Marine Sciences, College of Science, Shantou University, Shantou, Guangdong 515063, PR China.

International Journal of Biological Macromolecules
|November 23, 2021
PubMed
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This study presents a rapid chromatographic method for purifying Gracilaria corticata R-phycoerythrin (R-PE) using polyacrylamide gel electrophoresis. The purified R-PE demonstrates excellent stability and effective cellular imaging capabilities in vitro.

Area of Science:

  • Marine Biology
  • Biochemistry
  • Analytical Chemistry

Background:

  • Phycobiliproteins, such as R-phycoerythrin (R-PE), are vital light-harvesting pigments found in red algae.
  • Efficient purification methods are crucial for unlocking the potential of R-PE in various biotechnological applications.

Purpose of the Study:

  • To develop a rapid, single-step purification method for Gracilaria corticata R-phycoerythrin (R-PE).
  • To characterize the purified R-PE and assess its stability and cellular imaging properties.
  • To evaluate the feasibility of using R-PE for in vitro cell imaging applications.

Main Methods:

  • Purification of R-PE from Gracilaria corticata using polyacrylamide gel electrophoresis (PAGE).
  • Characterization of R-PE using UV-Vis spectroscopy, fluorescence spectroscopy, and SDS-PAGE.
Keywords:
Gracilaria corticataIn vitro cell imagingPhycobiliproteinPhysico-chemical propertiesR-phycoerythrinSDS-PAGE

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  • Assessment of R-PE stability under varying pH, light, and temperature conditions.
  • Evaluation of R-PE's cellular imaging potential on Vero and Hep-2 cell lines using fluorescence microscopy and flow cytometry.

    • Main Results:

    • A rapid chromatographic method yielded purified R-PE with a purity index of 4.2 and a recovery of 44.3%.
    • The purified R-PE exhibited characteristic spectral properties and was composed of α, β, and γ subunits (18, 21, and 31 kDa).
    • Optimal R-PE stability was observed at pH 7.0, 3000 lx light exposure for 6 hours, and 20°C.
    • R-PE demonstrated effective cellular imaging capabilities without compromising cell proliferation in vitro.

    Conclusions:

    • The developed PAGE-based method offers a time-efficient and effective approach for purifying phycobiliproteins like R-PE.
    • The purified R-PE possesses favorable stability and significant potential for applications in cellular imaging.
    • This study validates the utility of Gracilaria corticata R-PE as a fluorescent probe in biological research.