Purification and in situ immobilization of papain with aqueous two-phase system
| Autor: | Pengyong You, Xiangyu Gong, Ming Sun, Diansheng Xu, Erzheng Su, Mingliang Li, Dongzhi Wei |
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| Rok vydání: | 2010 |
| Předmět: |
Latex
Hydrolases lcsh:Medicine Plant Science Biochemistry Polyethylene Glycols Chitosan chemistry.chemical_compound Engineering Papain Biological Systems Engineering lcsh:Science Chromatography High Pressure Liquid Chromatography Multidisciplinary Aqueous solution Carica Enzyme Classes Physics Statistics Temperature Chemical Reactions Hydrogen-Ion Concentration Enzymes Chemistry Cross-Linking Reagents Electrophoresis Polyacrylamide Gel Research Article Biotechnology Immobilized enzyme Central composite design Materials Science Biophysics Bioengineering Protein Chemistry Catalysis Natural Materials Hydrolysis PEG ratio Biology lcsh:R Aqueous two-phase system Water Enzymes Immobilized chemistry Statistical Theories Biocatalysis lcsh:Q Plant Biotechnology Mathematics |
| Zdroj: | PLoS ONE PLoS ONE, Vol 5, Iss 12, p e15168 (2010) |
| ISSN: | 1932-6203 |
| Popis: | 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°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. |
| Databáze: | OpenAIRE |
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