Downstream processing of luciferase from fireflies (Photinus pyralis) using aqueous two-phase extraction |
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| Authors: | BS Priyanka Navin K Rastogi KSMS Raghavarao MS Thakur |
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| Institution: | 1. Department of Food Engineering, Central Food Technological Research Institute, Mysore 570020, Council of Scientific and Industrial Research, India;2. Department of Fermentation Technology and Bioengineering, Central Food Technological Research Institute, Mysore 570020, Council of Scientific and Industrial Research, India |
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| Abstract: | The firefly luciferase has been extensively used for sensitive detection of bacteria, gene expression and environmental toxins (biosensors). The aim of the present study was to design a simple and more efficient method for the purification and concentration of luciferase using aqueous two-phase extraction (ATPE). Downstream processing of luciferase from North American Firefly Photinus pyralis was carried out, for the first time, using polymer/salt aqueous two phase system (ATPS) at 4 °C. The enzyme was observed to preferentially partition to the polyethylene glycol (PEG) rich top phase. The best results of purification (13.69 fold) and enzyme activity recovery (118.34%) were observed in the system containing 4.0% (w/w) PEG (1500) and 20.5% (w/w) (NH4)2SO4 with a phase volume ratio of 0.21. |
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