Chapter 8
Analytical Protocols in Phycobiliproteins
Analysis
Milan R. Nikolic, Simeon Minic, Mirjana Macvanin,
Dragana Stanic-Vucinic, and Tanja Cirkovic Velickovic
Abstract The aim of this chapter is to review and discuss methodology and protocols
in the analysis of phycobiliproteins (phycocyanins, allophycocyanins, and phycoerythrins) and their chromophores. Due to the presence of multiple covalently bound
open-chain tetrapyrrole chromophores, phycobiliproteins are colored and strongly
fluorescent molecules, with high absorption coefficients (10
5 to 10
6 ) and excellent
fluorescent quantum yield (0.51 up to 0.98). Therefore, a vast number of methods
for phycobiliproteins analysis is based on these spectral characteristics, whereas
assessment of their bioactivity is related to their exceptional redox and metalchelating properties. This chapter is dedicated to methods used for isolation and
purification, structure analysis, physicochemical properties and stability characterization, quantification, as well as in vitro and in vivo biological activities evaluation. In addition, emerging approaches related to phycobiliproteins analysis are also
reviewed including interactions with other biomolecules and ions and identification
of phycobiliprotein (chromo)peptides by mass spectrometry.
M. R. Nikolic · S. Minic · D. Stanic-Vucinic · T. Cirkovic Velickovic (B)
Faculty of Chemistry, Center of Excellence for Molecular Food Sciences
and Department of Biochemistry, University of Belgrade, Belgrade, Serbia
e-mail: tcirkov@chem.bg.ac.rs
M. R. Nikolic
e-mail: mnikolic@chem.bg.ac.rs
S. Minic
e-mail: sminic@chem.bg.ac.rs
D. Stanic-Vucinic
e-mail: dstanic@chem.bg.ac.rs
M. Macvanin
Faculty of Chemistry, Center of Excellence for Molecular Food Sciences, University of Belgrade,
Belgrade, Serbia
e-mail: mirjana.macvanin@gmail.com
T. Cirkovic Velickovic
Serbian Academy of Sciences and Arts, Belgrade, Serbia
Ghent University Global Campus, Incheon, South Korea
Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
© Springer Nature Switzerland AG 2020
E. Jacob-Lopes et al. (eds.), Pigments from Microalgae Handbook,
https://doi.org/10.1007/978-3-030-50971-2_8
179
Analytical Protocols in Phycobiliproteins
Analysis
Milan R. Nikolic, Simeon Minic, Mirjana Macvanin,
Dragana Stanic-Vucinic, and Tanja Cirkovic Velickovic
Abstract The aim of this chapter is to review and discuss methodology and protocols
in the analysis of phycobiliproteins (phycocyanins, allophycocyanins, and phycoerythrins) and their chromophores. Due to the presence of multiple covalently bound
open-chain tetrapyrrole chromophores, phycobiliproteins are colored and strongly
fluorescent molecules, with high absorption coefficients (10
5 to 10
6 ) and excellent
fluorescent quantum yield (0.51 up to 0.98). Therefore, a vast number of methods
for phycobiliproteins analysis is based on these spectral characteristics, whereas
assessment of their bioactivity is related to their exceptional redox and metalchelating properties. This chapter is dedicated to methods used for isolation and
purification, structure analysis, physicochemical properties and stability characterization, quantification, as well as in vitro and in vivo biological activities evaluation. In addition, emerging approaches related to phycobiliproteins analysis are also
reviewed including interactions with other biomolecules and ions and identification
of phycobiliprotein (chromo)peptides by mass spectrometry.
M. R. Nikolic · S. Minic · D. Stanic-Vucinic · T. Cirkovic Velickovic (B)
Faculty of Chemistry, Center of Excellence for Molecular Food Sciences
and Department of Biochemistry, University of Belgrade, Belgrade, Serbia
e-mail: tcirkov@chem.bg.ac.rs
M. R. Nikolic
e-mail: mnikolic@chem.bg.ac.rs
S. Minic
e-mail: sminic@chem.bg.ac.rs
D. Stanic-Vucinic
e-mail: dstanic@chem.bg.ac.rs
M. Macvanin
Faculty of Chemistry, Center of Excellence for Molecular Food Sciences, University of Belgrade,
Belgrade, Serbia
e-mail: mirjana.macvanin@gmail.com
T. Cirkovic Velickovic
Serbian Academy of Sciences and Arts, Belgrade, Serbia
Ghent University Global Campus, Incheon, South Korea
Faculty of Bioscience Engineering, Ghent University, Ghent, Belgium
© Springer Nature Switzerland AG 2020
E. Jacob-Lopes et al. (eds.), Pigments from Microalgae Handbook,
https://doi.org/10.1007/978-3-030-50971-2_8
179