Priyanka, B. S. ; Rastogi, Navin K. ; Raghavarao, K. S. M. S. ; Thakur, M. S. (2012) Downstream processing of luciferase from fireflies (Photinus pyralis) using aqueous two-phase extraction Process Biochemistry, 47 (9). pp. 1358-1363. ISSN 1359-5113
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Official URL: http://www.sciencedirect.com/science/article/pii/S...
Related URL: http://dx.doi.org/10.1016/j.procbio.2012.04.022
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.
| Item Type: | Article |
|---|---|
| Source: | Copyright of this article belongs to Elsevier Science. |
| Keywords: | Downstream Processing; Luciferase; Bioseparation; Aqueous Two-Phase Extraction; Purification |
| ID Code: | 104300 |
| Deposited On: | 21 Dec 2017 11:55 |
| Last Modified: | 21 Dec 2017 11:55 |
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