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.

中文翻译：

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蛋白质色谱中的减少、再利用和再循环：利用废纸开发亲和吸附剂及其在鱼副产品中纯化蛋白酶中的应用。

在本研究中，我们报告了基于纤维素的亲和吸附剂的开发及其在从鱼副产品中纯化蛋白酶中的应用。亲和吸附剂以纤维素微纤维为基质合成，采用酸沉淀法从回收报纸中分离出来。使用三嗪染料 Cibacron Blue 3GA (CB3GA) 作为亲和配体，并将其直接固定在纤维素微纤维上。采用吸收平衡研究和正面亲和色谱法来评估吸附剂的色谱性能，使用牛血清白蛋白（BSA）和溶菌酶（LYS）作为模型蛋白。吸收平衡研究表明两种蛋白质的吸附均遵循朗缪尔等温线模型。吸附动力学服从准二阶模型。该亲和吸附剂用于开发黑鲷副产品（胃和胰腺）中蛋白酶的纯化程序。开发并优化了胰蛋白酶和糜蛋白酶的一步纯化方案。该方案提供了适合技术和工业目的的高产率酶。