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

Priyanka, B. S. and Rastogi, Navin K. and Raghavarao, K. S. M. S. and Thakur, M. S. (2013) Optimization of extraction of luciferase from fireflies (Photinus pyralis) using aqueous two-phase extraction. Separation and Purification Technology, 118. pp. 40-48. ISSN 1383-5866

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Abstract

A central composite rotatable design was employed to study the effect aqueous two-phase extraction conditions namely polyethylene glycol-1500 (PEG) concentration (12–16%), (NH4)2SO4 concentration (14–20%) and pH (6–9) on the extraction of luciferase from Photinus pyralis. Specific activity, activity recovery, purification fold, volume ratio, enzyme and protein partition coefficients were considered as response variables. The significant (p < 0.05) response surface models with high coefficients of determination values (R2) ranging from 0.89 to 0.95 were fitted for the experimental data, which indicated that the polynomial response models fitted well for describing the specific activity, activity recovery, purification fold, volume ratio, enzyme and protein partition coefficients. Based on the design, the optimal conditions were polyethylene glycol concentration 12%, (NH4)2SO4 concentration ranging from 16.48% to 17.86% and pH ranging from 8.17 to 8.43. The graphical optimization of superimposed contour plots fulfilled the conditions to obtain specific activity (Y1) ⩾ 5.5 × 106 RLU/mg, activity recovery (Y2) ⩾ 70%, purification fold (Y3) ⩾ 6.5, volume ratio (Y4) ⩽ 0.77, enzyme partition coefficient, Ke (Y5) ⩾ 95, protein partition coefficient, Kp (Y6) ⩽ 2.5. The study demonstrated that response surface methodology can be utilized for deriving the optimum conditions for optimizing the conditions for the aqueous two-phase extraction of luciferase.

Item Type:	Article
Additional Information:	Unmapped bibliographic data: DA - 2013/10/30/ [EPrints field already has value set] J2 - Separation and Purification Technology [Field not mapped to EPrints]
Uncontrolled Keywords:	Aqueous two-phase extraction, Downstream processing, Luciferase, Purification, Response surface methodology
Subjects:	600 Technology > 08 Food technology > 07 Food Engineering
Divisions:	Fermentation Technology and Bioengineering Food Engineering
Depositing User:	Food Sci. & Technol. Information Services
Date Deposited:	22 Sep 2016 11:17
Last Modified:	29 Dec 2016 13:13
URI:	http://ir.cftri.res.in/id/eprint/11231

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