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Published on: September 11, 2018
Processing-Independent Extracellular Production of High Purity C-Phycocyanin from Spirulina platensis
Anjali Purohit1, Varun Kumar1, Manisha Chownk1
1Center of Innovative and Applied Bioprocessing (CIAB), Sector-81, Knowledge City, Mohali 140306, Punjab, India.
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.
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