Purification, characterization and comparison of phycoerythrins from three different marine cyanobacterial cultures
BRD School of Biosciences, Sardar Patel Maidan, Vadtal Road, Satellite Campus, Post Box No. 39, Sardar Patel University, Vallabh Vidyanagar 388 120, Anand, Gujarat, India. Bioresource Technology
(Impact Factor: 4.49).
09/2010; 102(2):1795-802. DOI: 10.1016/j.biortech.2010.09.025
The present study is focused on purification, characterization and comparison of phycoerythrins from three different marine cyanobacterial cultures--hormidium sp. A27 DM, Lyngbya sp. A09 DM and Halomicronema sp. A32 DM. 'Phycoerythrin' was successfully purified and characterized. On SDS-PAGE, the PE purified from all three young cultures showed four bands--corresponding to α and β subunits of each of PE-I and PE-II. However, phycoerythrin purified after prolonged growth of Phormidium sp. A27 DM and Halomicronema sp. A32DM showed only one band corresponding to 14 kDa whereas Lyngbya sp. A09 DM continued to produce uncleaved phycoerythrin. The absorption spectra of purified PEs from all the three young and old cultures showed variations however the fluorescence studies of the purified PEs in all cases gave the emission spectra at around 580 nm. The described work is of great importance to understand the role of phycoerythrin in adapting cyanobacteria to stress conditions.
Available from: Rajeshwar P Sinha
- "Each subunit heterodimer has one to four (rings A, B, C and D) linear tetrapyrrole chromophores which are covalently attached to cysteines in the apoproteins with a thioether bond at the C-3 position on ring A and in some cases by an additional thioether bond at the C-18 position on ring D. The pathway of energy transfer within the subunits of PBPs starts from PE to PC to APC and finally reaches Chl a for photosynthesis[4,5]. Cyanobacteria can synthesize PBPs up to 50 % of its total soluble protein. "
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ABSTRACT: The wide applications of phycobiliproteins (PBPs) are essentially dependent on the purity ratio for the production of quality products. In this study, we present a novel method for the separation and purification of PBPs subunits such as phycocyanin (PC) and phycoerythrin (PE) from a rice-field cyanobacterium Nostoc sp. strain HKAR-11 using gel filtration and single step hydrophobic interaction chromatography. The separation of PBPs subunits was done using cycles of gel filtration chromatography (Sephacryl-S100 HR) with the addition of 0.5 M NaCl in 50 mM phosphate buffer. The final purification of PE and PC was performed using a HiTrap Phenyl FF column and yielded a quite high purity ratio of 11.53 and 5.75 with 83 and 73 % remaining concentrations for PE and PC, respectively. The purity of PE and PC was confirmed by emission fluorescence with excitation at 563 and 615 nm, respectively. The native molecular weight of purified PE (44 kDa) and PC (34 kDa) was determined by gel filtration chromatography. The purified PE and PC were analyzed by SDS-PAGE that showed the presence of only two bands of α and β subunits in comparison to the bands of crude PBPs. To the best of our knowledge, this is the most efficient methodology for obtaining quite high purity index of PC and PE from any rice-field cyanobacterial source.
Available from: Tonmoy Ghosh
- "A32DM, Pseudanabaena tenuis , Spirulina fusiformis , Arthronema africanum , Calothrix sp., Oscillatoria quadripunctulata , Pseudanabaena sp. Minkova et al. ( 2007 ), Santiago- Santos et al. ( 2004 ), Soni et al. ( 2010 ), Minkova et al. ( 2007 ), Mishra et al. ( 2008 ), Su et al. ( 2010 ), Cano-Europa et al. ( 2010 ), Parmar et al. ( 2011 ) and Mishra et al. ( 2011 ) Sonication Phycoerythrin Cyanosarcina sp. SK40, Phormidium sp. "
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ABSTRACT: Microalgae, one of the largest global primary producers, are a potential source of bioactive compounds. They are unique in producing superfine chemicals that can be used in various industrial sectors like pharmaceuticals, neutraceuticals and cosmeceuticals. The chapter is intended to provide an insight to two of the most important pigments obtained from them, phycobiliproteins and carotenoids having species specificity which can be used as a chemo- taxonomic marker. Their unique structural properties plays properties play a crucial role in their biological functions. The water soluble phycobiliproteins are used as fluorescent tags in flow cytometry and immunochemistry while liposoluble carotenoids are potential alternatives to synthetic dyes in the food industry.
Available from: Ravi Raghav Sonani
- "The filamentous cyanobacterium Phormidium rubidum A09DM (formally known as Lyngbya sp. A09DM) was isolated from sea shores of Okha, Gujarat, India, and identified by 16S rRNA gene sequence analysis (accession No. HM446280) (Parmar et al., 2011). The morphology of the alga was observed by using a light (BX41TF, Olympus Corporation, Tokyo, Japan) and scanning electron microscopy (Philips XL 30 ESEM). "
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ABSTRACT: The functionality and stability of phycobiliproteins (PBPs) phycoerythrin (PE), phycocyanin (PC) and allophycocyanin (APC) were investigated under various temperatures, pHs and oxidative stressors. All PBPs were thermostable up to 4-40°C; however, their concentration decreased rapidly at 60-80°C. The maximum stability of all PBPs was in the pH range 6.0-7.0. Decrease in PBPs content was found under high acidic (pH 2-4) and alkaline conditions (pH 8-12). The oxidizing agent (0.1-0.6%) showed the least effect on the stability of PBPs; however, 0.8-1.0% H2O2 caused significant loss of PBPs. Contrary to PE, PC and APC was more susceptible to an oxidizing agent. The chromophore associated with α- and β-subunit of PBPs and thus, their functionality (fluorescence) was severely affected under high temperature (60-80°C), and oxidizing agent, as well as low (2-4) and high (8-12) pH. Contrary to PC and APC, functionality of PE was surprisingly maintained even at pHs 6-12 and under oxidative stress.
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