Processing-Independent Extracellular Production of High Purity C-Phycocyanin from Spirulina platensis

Abstract

C-Phycocyanin (C-PC) is a natural blue colored protein of Spirulina platensis. Several methods based on chemical, physical, and enzymatic processes have been reported for the extraction of C-PC. However, most of the processes are either costly and/or time-consuming and/or produce C-PC with less purity. In view of this, a very simple and effective method was developed to extract C-PC with high purity and without involving any energy-consuming process from S. platensis. Wet biomass of S. platensis was harvested and incubated in an optimized 2-(N-morpholino)­ethanesulfonic acid buffer with no supplement of any other compounds under dark, anaerobic, and still conditions. These conditions facilitated the leaching out of C-PC from the cells. The purity ratio of the blue color C-PC supernatant obtained was enhanced by precipitation and dialysis. The purity ratio of C-PC produced was 0.644 and increased to 1.345 upon simple dialysis of the crude C-PC. The purity of C-PC was 6.17 upon purifying through column chromatography. Hence, a simple process was developed for producing C-PC of much higher purity than therapeutic grade (>4). Furthermore, the molecular regulation of the extracellular release of C-PC from S. platensis cells was monitored through relative expression levels of α and β subunits of C-PC genes during specific buffer vis-à-vis water as a control. Expression levels of genes encoding α and β subunits of C-PC were found to be upregulated in the case of the 2-(N-morpholino)­ethanesulfonic acid buffer treated algal cells compared to water treated algal cells. The higher level expression of these genes documented the higher and specific production of C-PC by the S. platensis upon 2-(N-morpholino)­ethanesulfonic acid buffer treatment.

WITHDRAWN: Spirulina C-phycocyanin: extraction, purification, stabilization, and inhibitory potential of key enzymes rRelated to pancreas inflammation, carbohydrate digestion, and glucose metabolism

“…The Sp was suspended in a phosphate buffer and subjected to multiple cycles of freezing (-20°C) and thawing (4°C) in the dark [21], resulting in the rupture of Sp cells and the release of CPC [22]. The extraction of CPC was carried out in the presence of 25 mM d 2-(N-morpholino)ethanesulfonic acid (MES) buffer (pH 6.5) [23], which was determined to be the optimal buffer for this purpose. The homogenate was then centrifuged at 200 rpm for 15 min to eliminate any remaining Sp [22].…”

“…Following the extraction and puri cation of betanin, the pure betanin and commercial betanin were subjected to infrared spectroscopy within the IR range of 3500 to 500 cm − 1. The FTIR spectra were analyzed to determine the position of functional groups, as per the methodology outlined in reference [23].…”

Ultrasound-Assisted Extraction and Assessment of Biological Activity of Phycobiliprotein-Rich Aqueous Extracts from Wild Cyanobacteria (Aphanizomenon flos-aquae)

“…For these reasons, we decided to include in our calculations the corrective formulas for A 620 and A 650 , as suggested by Lauceri et al Certainly, these equations are rather limited to Microcystis aeruginosa and do not consider other chlorophylls as well. Moreover, although various chromatographic techniques could be applied for further characterization and more accurate quantitative determination of the PBP content, spectrophotometric protocols as a result of their robustness, availability, low cost, and ease of operation remain the go-to choice for PBP quantification, as also depicted in the recent literature. − Nevertheless, we consider this approach as a step forward to a more accurate spectrophotometric determination of PC and APC contents upon development of various extraction protocols. The obtained results are discussed in the following sections.…”

Advances in delivery methods of Arthrospira platensis (spirulina) for enhanced therapeutic outcomes

“…Chlorophyll-a [65], C-phycocyanin (C-PC) [40,66,67], α-Glucans [68] and lastly a creative approach in using eutectic solvents for the extraction of natural products [69] for in vivo and in vitro studies.…”