Single Step Purification of Bromelain from Ananas Comosus Residues by PEG/Ammonium Sulphate Integrated Aqueous Two-Phase Systems

D. Coêlho, L. G. Spir, Letícia Celia de Lencastre Novaes, P. Mazzola, E. Silveira, E. Tambourgi, A. Pessoa
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引用次数: 1

Abstract

This work evaluates a single step purification of bromelain using a PEG/Ammonium Sulphate integrated aqueous two-phase systems (ATPS). This new process integrates the ammonium sulphate precipitation of unwanted proteins along with purification of target molecule by ATPS. The results, analysed by response surface methodology, showed that higher values of protein yield and enzyme activity recovery could be achieved by desirability optimization. Thus, it was possible to obtain a biological activity recovery over than 87.3% with a purification factor of 11.8 fold, using a system composed by 10.86% PEG 4,000 and 36.21% ammonium sulphate saturation. The purified bromelain showed an increased stability when supplemented with PEG, without any loss of activity after 7 hours of incubation
聚乙二醇/硫酸铵集成双水相体系单步纯化凤梨残渣中的菠萝蛋白酶
本研究评估了聚乙二醇/硫酸铵集成水两相系统(ATPS)对菠萝蛋白酶的单步纯化。这种新工艺结合了硫酸铵沉淀不需要的蛋白质和ATPS纯化的目标分子。响应面法分析结果表明,优化后的蛋白产量和酶活回收率均可达到较高的水平。因此,使用10.86% PEG 4000和36.21%硫酸铵饱和度组成的体系,可以获得超过87.3%的生物活性回收率和11.8倍的纯化系数。当添加PEG后,纯化的菠萝蛋白酶显示出更高的稳定性,在7小时的孵育后没有任何活性损失
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