Reduce, Reuse and Recycle in Protein Chromatography: Development of an Affinity Adsorbent from Waste Paper and Its Application for the Purification of Proteases from Fish By-Products

Abstract

In the present study, we report the development of a cellulose-based affinity adsorbent and its application for the purification of proteases from fish by-products. The affinity adsorbent was synthesized using cellulose microfibers as the matrix, isolated from recycled newspapers using the acid precipitation method. As an affinity ligand, the triazine dye Cibacron Blue 3GA (CB3GA) was used and immobilized directly onto the cellulose microfibers. Absorption equilibrium studies and frontal affinity chromatography were employed to evaluate the chromatographic performance of the adsorbent using as model proteins bovine serum albumin (BSA) and lysozyme (LYS). Absorption equilibrium studies suggest that the adsorption of both proteins obeys the Langmuir isotherm model. The kinetics of adsorption obey the pseudo-second-order model. The affinity adsorbent was applied for the development of a purification procedure for proteases from Sparus aurata by-products (stomach and pancreas). A single-step purification protocol for trypsin and chymotrypsin was developed and optimized. The protocol afforded enzymes with high yields suitable for technical and industrial purposes.

Recent trends in sorbents for bioaffinity chromatography

“…Hence, HSA was effectively isolated using the developed sorbent [121]. In recent studies, different dye ligands have been employed on different supports like nanofiber membranes [34], chitosan matrix [122], streamline matrix [37], cellulose microfibers [123], cryogel membrane [124], silica [125], and various polymers [121,126,127] for separation of different biological molecules. A schematic representation of the interaction between cibacron blue F3G-A as a typical dye ligand and albumin is shown in Figure 6 [121].…”

Prediction of the binding interactions between rosmarinic acid and cysteinyl leukotriene receptor type 1 by molecular docking and immobilized receptor chromatography

“…[1][2][3][4] Among these, the binding affinity investigations between the drugs and proteins have become the most interesting subject because of their crucial role in determining the pharmacological activity of the ligand. [5][6][7][8][9] Thus, increased attention has been drawn to the essential role of binding affinity investigations between drugs and proteins in discovering lead compounds.…”

Application of Biotechnological Techniques Aimed to Obtain Bioactive Compounds from Food Industry By-Products

“…Beyond vegetable by-products, the approaches aimed at valorizing animal food matrices such as meat and fish residues are also a current trend. Concerning that, Premetis and Labrou, 2020 [ 8 ], reported the development of a cellulose-based affinity adsorbent isolated from recycled newspapers and triazine dye Cibacron Blue 3GA as ligand able to purify proteases from fish by-products. The affinity adsorbent was applied for the development of a purification procedure for proteases from gilt-head bream ( Sparus aurata ) by-products such as stomach and pancreas.…”