Isolation, purification and properties of an R-Phycocyanin from the phycobilisomes of a marine red macroalga Polysiphonia urceolata

Phycobilisomes were prepared from a marine red macroalga Polysiphonia urceolata (P. urceolata) by sucrose step-gradient ultracentrifugation. From the prepared phycobilisomes, an R-phycocyanin was isolated by gel filtration on Sephadex G-150 and then purified by ion exchange chromatography on DEAE-Se...

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Published in:PloS one 2014-02, Vol.9 (2), p.e87833-e87833
Main Authors: Wang, Lu, Qu, Yanyan, Fu, Xuejun, Zhao, Mingri, Wang, Shumei, Sun, Li
Format: Article
Language:English
Subjects:
Algae
Anabaena
Analysis
Biology
China
Chromatography
Chromatography, Ion Exchange
Cyanobacteria
Dextrans
Electrophoresis, Polyacrylamide Gel
Energy
Filtration
Fingerprinting
Gel electrophoresis
Gel filtration
Isoelectric focusing
Laboratories
Life sciences
Light
Mass Spectrometry
Mass spectroscopy
Mastigocladus laminosus
Peptide mapping
Peptides - chemistry
pH effects
Phycobilisomes
Phycobilisomes - chemistry
Phycocyanin
Phycocyanin - chemistry
Phycocyanin - isolation & purification
Phycocyanins
Polypeptides
Polysiphonia urceolata
Purification
Rhodophyta
Rhodophyta - chemistry
Seaweeds
Sodium lauryl sulfate
Spectrometry, Fluorescence
Spectroscopy
Sucrose
Sugar
Synechococcus
Ultracentrifugation
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creator Wang, Lu
Qu, Yanyan
Fu, Xuejun
Zhao, Mingri
Wang, Shumei
Sun, Li
description Phycobilisomes were prepared from a marine red macroalga Polysiphonia urceolata (P. urceolata) by sucrose step-gradient ultracentrifugation. From the prepared phycobilisomes, an R-phycocyanin was isolated by gel filtration on Sephadex G-150 and then purified by ion exchange chromatography on DEAE-Sepharose Fast Flow and native polyacrylamide gel electrophoresis (PAGE) performed in neutral buffer systems. The purified R-phycocyanins showed not only a homogeneous trimer of 136 kDa in gel filtration and a single band in native PAGE, but also exhibited one band at about pH 5.7 in native isoelectric focusing (IEF). By a gradient SDS-PAGE the purified R-phycocyanin was determined to contain one α subunit of 17.5 kDa (α (17.5)) and two β subunits of 21.3 kDa and 22.6 kDa (β (21.3) and β (22.6)). The analysis from denaturing isoelectric focusing and two-dimension PAGE demonstrated that α (17.5), β (21.3) and β (22.6) had their pIs of 6.4, 5.3 and 5.4, respectively. Furthermore, mass spectroscopy analysis of β (21.3) and β (22.6) by MALDI-TOF mass spectrometry demonstrated the two β subunits had differences in peptide mass fingerprinting. These results revealed that the prepared R-phycocyanins were composed of one α and two β subunits. and , which have a structural foundation to show their pIs too close for them to be definitely resolved by native IEF, are postulated to be the most possible trimeric forms of the R-phycocyanins prepared from the phycobilisomes of P. urceolata.
doi_str_mv 10.1371/journal.pone.0087833
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These results revealed that the prepared R-phycocyanins were composed of one α and two β subunits. and , which have a structural foundation to show their pIs too close for them to be definitely resolved by native IEF, are postulated to be the most possible trimeric forms of the R-phycocyanins prepared from the phycobilisomes of P. urceolata.</description><subject>Algae</subject><subject>Anabaena</subject><subject>Analysis</subject><subject>Biology</subject><subject>China</subject><subject>Chromatography</subject><subject>Chromatography, Ion Exchange</subject><subject>Cyanobacteria</subject><subject>Dextrans</subject><subject>Electrophoresis, Polyacrylamide Gel</subject><subject>Energy</subject><subject>Filtration</subject><subject>Fingerprinting</subject><subject>Gel electrophoresis</subject><subject>Gel filtration</subject><subject>Isoelectric focusing</subject><subject>Laboratories</subject><subject>Life sciences</subject><subject>Light</subject><subject>Mass Spectrometry</subject><subject>Mass spectroscopy</subject><subject>Mastigocladus laminosus</subject><subject>Peptide mapping</subject><subject>Peptides - chemistry</subject><subject>pH effects</subject><subject>Phycobilisomes</subject><subject>Phycobilisomes - chemistry</subject><subject>Phycocyanin</subject><subject>Phycocyanin - chemistry</subject><subject>Phycocyanin - isolation &amp; 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From the prepared phycobilisomes, an R-phycocyanin was isolated by gel filtration on Sephadex G-150 and then purified by ion exchange chromatography on DEAE-Sepharose Fast Flow and native polyacrylamide gel electrophoresis (PAGE) performed in neutral buffer systems. The purified R-phycocyanins showed not only a homogeneous trimer of 136 kDa in gel filtration and a single band in native PAGE, but also exhibited one band at about pH 5.7 in native isoelectric focusing (IEF). By a gradient SDS-PAGE the purified R-phycocyanin was determined to contain one α subunit of 17.5 kDa (α (17.5)) and two β subunits of 21.3 kDa and 22.6 kDa (β (21.3) and β (22.6)). The analysis from denaturing isoelectric focusing and two-dimension PAGE demonstrated that α (17.5), β (21.3) and β (22.6) had their pIs of 6.4, 5.3 and 5.4, respectively. Furthermore, mass spectroscopy analysis of β (21.3) and β (22.6) by MALDI-TOF mass spectrometry demonstrated the two β subunits had differences in peptide mass fingerprinting. These results revealed that the prepared R-phycocyanins were composed of one α and two β subunits. and , which have a structural foundation to show their pIs too close for them to be definitely resolved by native IEF, are postulated to be the most possible trimeric forms of the R-phycocyanins prepared from the phycobilisomes of P. urceolata.</abstract><cop>United States</cop><pub>Public Library of Science</pub><pmid>24504114</pmid><doi>10.1371/journal.pone.0087833</doi><tpages>e87833</tpages><oa>free_for_read</oa></addata></record>
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subjects Algae
Anabaena
Analysis
Biology
China
Chromatography
Chromatography, Ion Exchange
Cyanobacteria
Dextrans
Electrophoresis, Polyacrylamide Gel
Energy
Filtration
Fingerprinting
Gel electrophoresis
Gel filtration
Isoelectric focusing
Laboratories
Life sciences
Light
Mass Spectrometry
Mass spectroscopy
Mastigocladus laminosus
Peptide mapping
Peptides - chemistry
pH effects
Phycobilisomes
Phycobilisomes - chemistry
Phycocyanin
Phycocyanin - chemistry
Phycocyanin - isolation & purification
Phycocyanins
Polypeptides
Polysiphonia urceolata
Purification
Rhodophyta
Rhodophyta - chemistry
Seaweeds
Sodium lauryl sulfate
Spectrometry, Fluorescence
Spectroscopy
Sucrose
Sugar
Synechococcus
Ultracentrifugation
title Isolation, purification and properties of an R-Phycocyanin from the phycobilisomes of a marine red macroalga Polysiphonia urceolata
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