Downstream processing of luciferase from fireflies (Photinus pyralis) using aqueous two-phase extraction

作者：Priyanka B S; Rastogi Navin K; Raghavaraoar K S M S^*; Thakur M S

来源：Process Biochemistry, 2012, 47(9): 1358-1363.

DOI：10.1016/j.procbio.2012.04.022

• Downstream processing
• Luciferase
• Bioseparation
• Aqueous two-phase extraction
• Purification

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 degrees 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)(2)SO4 with a phase volume ratio of 0.21.

• 出版日期2012-9