molecular diagnose due to its strong fluorescence. However, the
industrial production of PC is limited due to challenging and
unstable purification process [6]. Appropriate control of temperature, pH, and ionic strength during the extraction and purification
of PC is important for the stability of PC. Consequently, an efficient, eco-friendly, and economical PC purification methods is
essential to attain the desired yield, quality, and purity and also to
preserve its properties [7]. A number of methods for extraction and
purification of PC from cyanobacteria have been reported in the
literature [6, 8].
The selection of suitable cell disruption method and appropriate buffer for phycocyanin extraction are also very crucial factors for
obtaining the high yield and purity. Several cell disruption methods
including mechanical, chemical, and enzymatic disruption have
been used for PC extraction from wet/dry biomass of cyanobacteria [8–11]. The efficiency of cell disruption method depends on
cell wall characteristics of species and operating conditions. Generally, mechanical disruption is the preferred method for PC extraction because of high cell disruption efficiency compared with other
methods, and also it avoids chemical contamination. Methods
reported for the purification of PC from cyanobacteria involve the
combination of chromatographic and non-chromatographic methods such as ultrafiltration, aqueous two-phase extraction, expanded
bed chromatography, gel filtration, and ion-exchange chromatography [11–14]. Cost, complexity, and selectivity are the main factors that should be considered while choosing suitable purification
method for PC. This chapter describes simple, economical, and
effective procedure to extract and purify analytical grade PC from
cyanobacteria using three-step sequential method consisting of
extraction of PC, fractional ammonium sulfate precipitation, and
chromatographic purification by using sephadex G-25 gel filtration
and DEAE-sephadex ion-exchange column chromatography.
2 Materials
2.1 Equipments
1. Ultrasonic homogenizer.
2. Refrigerated centrifuge.
3. UV-visible spectrophotometer.
4. Vortex.
5. Magnetic stirrer.
2.2 Extraction Buffer
Na-phosphate buffer (50 mM, pH 7.0): Weigh 3.29 g of NaH 2PO 4 ·2H 2 O and 7.73 g of Na 2 HPO 4 ·7H 2 O, and transfer to the 1 L
glass beaker containing 250 mL deionized water. Mix well to
dissolve salts completely. Further, add water to a volume of
174
Mahammed Ilyas Khazi et al.
industrial production of PC is limited due to challenging and
unstable purification process [6]. Appropriate control of temperature, pH, and ionic strength during the extraction and purification
of PC is important for the stability of PC. Consequently, an efficient, eco-friendly, and economical PC purification methods is
essential to attain the desired yield, quality, and purity and also to
preserve its properties [7]. A number of methods for extraction and
purification of PC from cyanobacteria have been reported in the
literature [6, 8].
The selection of suitable cell disruption method and appropriate buffer for phycocyanin extraction are also very crucial factors for
obtaining the high yield and purity. Several cell disruption methods
including mechanical, chemical, and enzymatic disruption have
been used for PC extraction from wet/dry biomass of cyanobacteria [8–11]. The efficiency of cell disruption method depends on
cell wall characteristics of species and operating conditions. Generally, mechanical disruption is the preferred method for PC extraction because of high cell disruption efficiency compared with other
methods, and also it avoids chemical contamination. Methods
reported for the purification of PC from cyanobacteria involve the
combination of chromatographic and non-chromatographic methods such as ultrafiltration, aqueous two-phase extraction, expanded
bed chromatography, gel filtration, and ion-exchange chromatography [11–14]. Cost, complexity, and selectivity are the main factors that should be considered while choosing suitable purification
method for PC. This chapter describes simple, economical, and
effective procedure to extract and purify analytical grade PC from
cyanobacteria using three-step sequential method consisting of
extraction of PC, fractional ammonium sulfate precipitation, and
chromatographic purification by using sephadex G-25 gel filtration
and DEAE-sephadex ion-exchange column chromatography.
2 Materials
2.1 Equipments
1. Ultrasonic homogenizer.
2. Refrigerated centrifuge.
3. UV-visible spectrophotometer.
4. Vortex.
5. Magnetic stirrer.
2.2 Extraction Buffer
Na-phosphate buffer (50 mM, pH 7.0): Weigh 3.29 g of NaH 2PO 4 ·2H 2 O and 7.73 g of Na 2 HPO 4 ·7H 2 O, and transfer to the 1 L
glass beaker containing 250 mL deionized water. Mix well to
dissolve salts completely. Further, add water to a volume of
174
Mahammed Ilyas Khazi et al.