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通过非传统的双水相体系(PEG/硫酸铵)对菠萝蛋白酶进行纯化。

Bromelain purification through unconventional aqueous two-phase system (PEG/ammonium sulphate).

机构信息

Departamento de Engenharia de Sistemas Químicos, Faculdade de Engenharia Química, Universidade Estadual de Campinas, Cidade Universitária "Zeferino Vaz", São Paulo, Brazil.

出版信息

Bioprocess Biosyst Eng. 2013 Feb;36(2):185-92. doi: 10.1007/s00449-012-0774-5. Epub 2012 Jul 5.

Abstract

This paper focuses on the feasibility of unconventional aqueous two-phase systems for bromelain purification from pineapple processing waste. The main difference in comparison with conventional systems is the integration of the liquid-liquid extraction technique with fractional precipitation, which can decrease the protein content with no loss of biological activity by removing of unwanted molecules. The analysis of the results was based on the response surface methodology and revealed that the use of the desirability optimisation methodology (DOM) was necessary to achieve higher purification factor values and greater bromelain recovery. The use of DOM yielded an 11.80-fold purification factor and 66.38 % biological activity recovery using poly(ethylene glycol) (PEG) with a molar mass of 4,000, 10.86 % PEG concentration (m/m) and 36.21 % saturation of ammonium sulphate.

摘要

本文研究了非传统双水相体系在菠萝加工废弃物中菠萝蛋白酶纯化的可行性。与传统体系的主要区别在于,液-液萃取技术与分级沉淀相结合,可以通过去除不需要的分子,在不损失生物活性的情况下降低蛋白质含量。结果分析基于响应面法,结果表明,需要使用理想优化方法 (DOM) 才能获得更高的纯化因子值和更大的菠萝蛋白酶回收率。使用摩尔质量为 4000 的聚乙二醇 (PEG)、10.86% (m/m) 的 PEG 浓度和 36.21% 的硫酸铵饱和度,通过 DOM 得到了 11.80 倍的纯化因子和 66.38% 的生物活性回收率。

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