使用响应面法优化水相两相系统中酸性蛋白酶分配和纯化的条件。
Optimization of conditions for acid protease partitioning and purification in aqueous two-phase systems using response surface methodology.
作者信息
Pericin Draginja M, Madarev-Popović Senka Z, Radulović-Popović Ljiljana M
机构信息
Department of Applied and Engineering Chemistry, Faculty of Technology, University of Novi Sad, Bulevar Cara Lazara 1, Novi Sad, Serbia.
出版信息
Biotechnol Lett. 2009 Jan;31(1):43-7. doi: 10.1007/s10529-008-9830-2. Epub 2008 Sep 5.
Aspergillopepsin I, an acid protease, was purified using an aqueous two-phase system that comprised various combinations of polyethylene glycol (PEG), NaH2PO4 and NaCl. Partition of the enzyme depended upon the molecular mass of the PEG and the presence of NaCl. With PEG 1500, 4000 and 6000, the partition coefficients were increased by 1,500-, 1,800- and 560-fold compared to values without NaCl. The presence of NaCl (8.75%, w/w) increased purification by 3.8, 9.5 and 2.8 times into these respective PEGs. The optimal aqueous two-phase system for acid protease purification was developed using response surface methodology. This system contained 17.3% of PEG 4000 (w/w), 15% NaH2PO4 (w/w) and 8.75% NaCl (w/w) and provided the best partition coefficient (Ke > 1,100) and yield over 99% in the same phase. The optimal ATPS purification factor of acid protease was over 5.
曲霉胃蛋白酶I是一种酸性蛋白酶，使用由聚乙二醇（PEG）、NaH2PO4和NaCl的各种组合构成的双水相系统进行纯化。该酶的分配取决于PEG的分子量和NaCl的存在。对于PEG 1500、4000和6000，与不存在NaCl时的值相比，分配系数分别增加了1500倍、1800倍和560倍。NaCl（8.75%，w/w）的存在使进入这些相应PEG的纯化倍数分别提高了3.8倍、9.5倍和2.8倍。使用响应面法开发了用于酸性蛋白酶纯化的最佳双水相系统。该系统含有17.3%的PEG 4000（w/w）、15%的NaH2PO4（w/w）和8.75%的NaCl（w/w），并在同一相中提供了最佳分配系数（Ke>1100）和超过99%的产率。酸性蛋白酶的最佳双水相系统纯化因子超过5。
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