Abstract
Ribulose-1,5-biphosphate carboxylase/oxygenase (RuBisCO) is the most abundant protein on the planet, being present in plants, algae and various species of bacteria, with application in the pharmaceutical, chemical, cosmetic and food industries. However, current extraction methods of RuBisCO do not allow high yields of extraction. Therefore, the development of an efficient and selective RuBisCOs’ extraction method is required. In this work, aqueous solutions of biocompatible ionic liquids (ILs), i.e., ILs derived from choline and analogues of glycine-betaine, were applied in the RuBisCO’s extraction from spinach leaves. Three commercial imidazolium-based ILs were also investigated for comparison purposes. To optimize RuBisCO’s extraction conditions, response surface methodology was applied. Under optimum extraction conditions, extraction yields of 10.92 and 10.57 mg of RuBisCO/g of biomass were obtained with the ILs cholinium acetate ([Ch][Ac]) and cholinium chloride ([Ch]Cl), respectively. Circular dichroism (CD) spectroscopy results show that the secondary structure of RuBisCO is better preserved in the IL solutions when compared to the commonly used extraction solvent. The obtained results indicate that cholinium-based ILs are a promising and viable alternative for the extraction of RuBisCO from vegetable biomass.
Highlights
Ribulose-1,5-biphosphate carboxylase/oxygenase, RuBisCO (EC 4.1.1.39), is the most abundant protein and is responsible for the atmospheric carbon fixation in photosynthetic organisms, such as plants, algae and various species of bacteria [1,2,3,4,5]
The influence of the three independent variables on the concentration of RuBisCO in the extracts obtained are shown in Figure 5A–C and in the Supplementary Materials, Figure S8 and Tables S7 and S8
ionic liquids (ILs) aqueous solutions were investigated as alternative solvent aiming at developing a more sustainable approach for RuBisCO’s extraction from spinach leaves
Summary
Ribulose-1,5-biphosphate carboxylase/oxygenase, RuBisCO (EC 4.1.1.39), is the most abundant protein (enzyme) and is responsible for the atmospheric carbon fixation in photosynthetic organisms, such as plants, algae and various species of bacteria [1,2,3,4,5]. RuBisCO can be applied in several fields, such as food and feed, chemical, pharmaceutical and cosmetic industries [1,6,7,8,9,10,11,12,13,14,15]. RuBisCO is already employed as a protein source through the incorporation of a white protein concentrate, which consists of soluble leaf proteins [4,11]. RuBisCO gels (products from thermal denaturation) have the potential to be used in food formulation [12], while undenatured RuBisCO can be applied in foaming and emulsifying and to form gels [12]. On the other hand, when addressing applications such as cosmetics and pharmaceuticals, high-purity RuBisCO is demanded
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