文献类型：期刊文献

英文题名：Purification and In Situ Immobilization of Papain with Aqueous Two-Phase System

作者：Li, Mingliang[1];Su, Erzheng[1];You, Pengyong[1];Gong, Xiangyu[1];Sun, Ming[1];Xu, Diansheng[1];Wei, Dongzhi[1]

机构：[1]E China Univ Sci & Technol, New World Inst Biotechnol, State Key Lab Bioreactor Engn, Shanghai 200237, Peoples R China

年份：2010

卷号：5

期号：12

外文期刊名：PLOS ONE

收录：;WOS:【SCI-EXPANDED(收录号:WOS:000285246900032)】；

基金：This work was supported by the National Basic Research Program (973 Program) of China (2007CB714300), the National Special Fund for State Key Laboratory of Bioreactor Engineering (Grant No. 2060204) and the Specialized Research Fund for the Doctoral Program (New Teachers) of Higher Education (20090074120015). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

语种：英文

摘要：Papain was purified from spray-dried Carica papaya latex using aqueous two-phase system (ATPS). Then it was recovered from PEG phase by in situ immobilization or preparing cross-linked enzyme aggregates (CLEAs). The Plackett-Burman design and the central composite design (CCD) together with the response surface methodology (RSM) were used to optimize the APTS processes. The highly purified papain (96-100%) was achieved under the optimized conditions: 40% (w/w) 15 mg/ml enzyme solution, 14.33-17.65% (w/w) PEG 6000, 14.27-14.42% (w/w) NaH2PO4/K2HPO4 and pH 5.77-6.30 at 20 degrees C. An in situ enzyme immobilization approach, carried out by directly dispersing aminated supports and chitosan beads into the PEG phase, was investigated to recover papain, in which a high immobilization yield (>90%) and activity recovery (>40%) was obtained. Moreover, CLEAs were successfully used in recovering papain from PEG phase with a hydrolytic activity hundreds times higher than the carrier-bound immobilized papain.