Single-step purification of C-phycocyanin from Arthrospira platensis using aqueous two-phase system based on natural dee
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Single-step purification of C-phycocyanin from Arthrospira platensis using aqueous two-phase system based on natural deep eutectic solvents Quanfu Wang 1
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Nana Wei 1 & Yifan Wang 1 & Yanhua Hou 1
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Xiulian Ren 1 & Qifeng Wei 1
Received: 12 December 2019 / Revised and accepted: 18 August 2020 # Springer Nature B.V. 2020
Abstract Natural deep eutectic solvents (NADESs) are an emerging type of green ionic liquid that has been used in aqueous two-phase systems (ATPSs). C-phycocyanin (C-PC) is an important fluorescent protein found in cyanobacteria with multiple applications. In this study, NADES [choline chloride-urea, ChCl-U]-ATPS (NADES-ATPS) was screened as an optimum system for extracting C-PC from Arthrospira platensis. Under the optimal purification conditions (crude extracted C-PC, 500 μg; [ChClU], 3.25 g; K2HPO4, 0.90 g mL−1; extraction time, 2 h; NaCl, 2% and 25 °C), C-PC with a purity and recovery of 3.383 and 65.64%, respectively, was obtained by single-step extraction. The purity of C-PC could increase to 4.100 after four extraction steps. The purified C-PC was further analyzed via sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), which showed α and β subunits with molecular weights of 18.0 and 21.0 kDa, respectively. Importantly, the NADES can successfully be recycled and reused for at least five pretreatment cycles. Furthermore, no structural changes in C-PC were detected, and the aggregation phenomenon of NADES with C-PC may be the extraction mechanism. Such a green, easily synthesized, recyclable and efficient method using NADES-ATPS could hold promise for wide applications for the purification of C-PC and other proteins. Keywords Ionic liquids . Natural deep eutectic solvents . Aqueous two-phase system . C-phycocyanin . Recyclable
Introduction Arthrospira platensis, previously known as Spirulina platensis, is a cyanobacterium that has attracted attention due to its nutritional value and high economical value. Cphycocyanin (C-PC) comprises as much as 40–60% of the total soluble protein in cells. Under natural conditions, CPC with a strong red fluorescence contains α and β subunits expressed as (αβ)3 or (αβ)6 aggregates (RodríguezSánchez et al. 2012). C-PC has been widely applied as an antioxidant (Patel et al. 2018), immune enhancer, and antitumor agent that induces apoptosis and inhibit cell proliferation (Cian et al. 2012; Saini and Sanyal 2015). In our previous study, strong fluorescence of C-PC can be processed into a fluorescent probe to detect mercury ions in
* Yanhua Hou [email protected] 1
School of Marine Science and Technology, Harbin Institute of Technology, Weihai 264209, People’s Republic of China
seafood (Hou et al. 2017) and fluorescent reagents in medical diagnostic bioengineering and other fields. Therefore, as a natural coloring and safety agent, C-PC with high purity can be extensively used in industrial fields such as food, cosmetics, and pharmaceuticals (Abd El Baky et al. 2015; Kovač et al. 2017; Figueira et al. 2018; Mogany et al. 2019). In view of t
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