Efficient Extraction of the RuBisCO Enzyme from Spinach Leaves Using Aqueous Solutions of Biocompatible Ionic Liquids

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.

Towards the sustainable extraction and purification of non-animal proteins from biomass using alternative solvents

“…Br). 22 The authors applied a response surface methodology (RSM) to optimize the conditions for RuBisCO extraction. Under these conditions, extraction yields of 10.92 and 10.57 mg of RuBisCO per g of biomass were obtained with [Ch][Ac] and [Ch] Cl, respectively.…”

Enabling Sustainable Chemistry with Ionic Liquids and Deep Eutectic Solvents: A Fad or the Future?

“…As components of the human diet, these bioderived ILs and DESs eliminate the risk of immunogenicity, which is possible if the bacteria‐derived protein A, G, A/G, or L ligand leaches and contaminates the IgG during ligand‐based affinity chromatographic purification. Choline, amino acids, and fatty acids exemplify these bioderived components of DESs increasingly used to extract and purify biomolecules [188, 136, 198–204] . Various bioderived ILs combined with poly(propylene glycol) extract IgG from rabbit serum into the IL‐rich phase, with the recovery yield reaching 85 % [205] .…”

“…Various bioderived ILs combined with poly(propylene glycol) extract IgG from rabbit serum into the IL‐rich phase, with the recovery yield reaching 85 % [205] . The purified antibody or protein preserves its native structure without denaturation or degradation better in IL‐based ABSs than in conventional extraction solvents [198, 201] …”

Valorisation of red beet waste: one-step extraction and separation of betalains and chlorophylls using thermoreversible aqueous biphasic systems

“…11 Moreover, aqueous solutions of ILs have shown a remarkable performance in the extraction of a plethora of biomolecules, such as alkaloids and flavonoids from complex sources like biomass. 12…”

“…11 Moreover, aqueous solutions of ILs have shown a remarkable performance in the extraction of a plethora of biomolecules, such as alkaloids and flavonoids from complex sources like biomass. 12 Biomass is a complex matrix, requiring several unit operations to extract and separate high-value compounds. However, there is a high demand to develop integrated extraction-separation processes, or stimuli-responsive ones, combining multiple unit operations that could make them more economical and sustainable.…”