C-Phycocyanin from Oscillatoria tenuis exhibited an antioxidant and in vitro antiproliferative activity through induction of apoptosis and G0/G1 cell cycle arrest
Graphical abstract
Highlights
► Efficient and low-cost method developed for purification of C-phycocyanin (CPC) from Oscillatoria tenuis. ► The purity ratio of CPC was 4.88, which is analytical grade value for pure CPC. ► CPC from O. tenuis potentially scavenges free radicals. ► CPC from O. tenuis can also potentially inhibit cancer cell growth in vitro. ► CPC induces apoptotic characteristics of membrane blebs, apoptosis and G0/G1 cell cycle arrest.
Introduction
Chemoprevention is an effective way to reduce cancer risk. Natural products have been the mainstay of cancer chemotherapy for the past 30 years. Blue-green algae are the most primitive life forms on earth with nutrient-dense, edible forms like Nostoc, Spirulina, Aphanizomenon species, etc. The cyanobacterial phycocyanin (CPC) is the major phycobiliprotein in cyanobacteria. This blue colour red fluorescing biliprotein was first reported in 1928 by Lemberg (Patil et al., 2006). It consist of a protein component and a chromophore, and the protein moiety consists of α and β subunit (Patil et al., 2006). Phycocyanin is a natural blue colourant, has uses as a food colourant for chewing gum, sorbets, soft drinks, candies and cosmetics, including lipstick and eyeliners. Small quantities are also used as biochemical tracers in immunoassays due to its fluorescent properties (Silveira, Burket, Costa, Burket, & Kalil, 2007).
Phycocyanin has been proven to have therapeutic properties including antioxidant, anti-inflammatory and anti-cancer activities (Eriksen, 2008, Madhyastha et al., 2009). Recent studies have demonstrated antioxidant, anti-mutagenic, antiviral, anticancer, anti-allergic, immune enhancing, hepatoprotective, blood vessel-relaxing and blood lipid-lowering effects of Spirulina extracts. The biological and pharmacological properties of Spirulina were attributed mainly to calcium-spirulan and CPC (Subhashini et al., 2004). The cost of phycocyanin products varies widely and is dependent on the purity ratio, which is defined as the ratio of absorbance at 620 and 280 nm (A620/A280).
CPC has been purified using a number of combinations of chromatographic steps such as ion exchange chromatography, gel filtration chromatography, chromatography on hydroxyapatite and expanded bed adsorption chromatography (Bermejo et al., 2003, Santiago-Santos et al., 2004, Soni et al., 2006, Wang, 2002). In most cases, it has been purified by sequential application of either cation or anion exchange chromatography, followed by size exclusion chromatography as a final polishing step. Though purification was achieved in the above methods, high purity methods were very expensive while low-cost methods were time-consuming. Moreover, the combination of several steps decreases the yield considerably, requiring large culture volumes for purification (Jobby & Sharma, 2003). Hence the present study attempted to simplify the purification process as well as reduce the cost of process. Additionally we were interested to study the biological activities of CPC from Oscillatoria tenuis.
Section snippets
Chemicals and reagents
Seralose 6B gel filtration column, Griess reagent and ascorbic acid were purchased from SRL Chemicals, Mumbai, India. 1,1-Diphenyl-2-picrylhydrazyl (DPPH) and Dulbecco’s Modified Eagles Medium (DMEM) were purchased from Hi Media, Mumbai, India. Nitroblue tetrazolium (NBT), nicotinamide adenine dinucleotide (NADH), (4-(2-hydroxyethyl)-1-piperazineethane sulphonic acid) (HEPES), penicillin, streptomycin, propidium iodide, ethylenediaminetetraacetic acid (EDTA), WST-8 kit, and dimethyl sulphoxide
Purification and characterisation of CPC from O. tenuis
The extraction of phycobiliproteins in O. tenuis revealed CPC as the major component with respect to other phycobiliproteins, such as APC and PE, which makes the purification process easy as well as efficient (Fig. 1 and Table 1). Because PE is present in very low amount, this molecule does not interfere in the purification process. This may be due to O. tenuis specificity in the phycobiliprotein content and the culture conditions.
During the process of extraction, cyanobacterial cells were
References (33)
- et al.
Preparative purification of B-phycoerythrin from the microalga Porphyridium cruentum by expanded-bed adsorption chromatography
Journal of Chromatography B
(2003) - et al.
Biological activities of sulfated polysaccharides from tropical seaweeds
Biomedicine & Pharmacotherapy
(2010) - et al.
Action of phenolic derivatives (acetaminophen, salicylate and 5-aminosalicylate) as inhibitors of membrane lipid peroxidation and as peroxyl radical scavengers
Archives of Biochemistry and Biophysics
(1994) - et al.
Effects of non-steroidal anti-inflammatory drugs on proliferation and on induction of apoptosis in colon cancer cells by a prostaglandin-independent pathway
Biochemical Pharmacology
(1996) - et al.
Rapid purification of recombinant βB2-crystallin using hydrophobic interaction chromatography
Protein Expression and Purification
(2003) - et al.
Free radical-scavenging and inhibition of nitric oxide production by four grades of pine mushroom (Tricholoma matsutake Sing)
Food Chemistry
(2007) - et al.
One-step chromatography method for efficient separation and purification of R-phycoerythrin from Polysiphonia urceolata
Journal of Biotechnology
(2005) - et al.
C-phycocyanin from Spirulina fussiformis exposed to blue light demonstrates higher efficacy of in vitro antioxidant activity
Biochemical Engineering Journal
(2009) - et al.
Propolis induces cell cycle arrest and apoptosis in human leukemic U937 cells through Bcl-2/Bax regulation
Environmental Toxicology and Pharmacology
(2008) - et al.
Purification and characterization of C-phycocyanin from cyanobacterial species of marine and freshwater habitat
Protein Expression and Purification
(2005)
C-Phycocyanin, a selective cyclooxygenase-2 inhibitor, induces apoptosis in lipopolysaccharide-stimulated RAW 264.7 macrophages
Biochemical and Biophysical Research Communications
Isolation and characterization of macromolecular protein R-Phycoerythrin from Portieria hornemannii
International Journal of Biological Macromolecules
Optimization of phycocyanin extraction from Spirulina platensis using factorial design
Bioresource Technology
Extraction, purification and characterization of phycocyanin from Oscillatoria quadripunctulata – Isolated from the rocky shores of Bet-Dwarka, Gujarat, India
Process Biochemistry
Molecular mechanisms in C-Phycocyanin induced apoptosis in human chronic myeloid leukemia cell line-K562
Biochemical Pharmacology
A novel disintegrin protein from Naja naja venom induces cytotoxicity and apoptosis in human cancer cell lines in vitro
Process Biochemistry
Cited by (109)
Bioactive C-phycocyanin exerts immunomodulatory and antitumor activity in mice with induced melanoma
2024, Toxicology and Applied PharmacologyProperties and potential applications of bioconjugates of R-phycoerythrin with Ag° or CdS nanoparticle synthesized in its tunnel cavity: A review
2024, International Journal of Biological MacromoleculesCyanobacterial interactions and symbiosis
2023, Cyanobacteria: Metabolisms to Molecules