Comparison of Purification Methods to Purify Recombinant Bromelain from Escherichia coli BL21-A1
Recombinant bromelain is a cysteine protease that can be exploited for its protease activity in food and pharmaceutical applications. The aim of this study was to compare different purification methods aqueous two-phase system (ATPS), ammonium sulphate precipitation,...
| Main Authors: | , , , , , , |
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| Format: | Article |
| Language: | English |
| Published: |
The Malaysian Analytical Sciences Society
2017
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| Subjects: | |
| Online Access: | http://umpir.ump.edu.my/id/eprint/19928/ http://umpir.ump.edu.my/id/eprint/19928/ http://umpir.ump.edu.my/id/eprint/19928/ http://umpir.ump.edu.my/id/eprint/19928/1/fkksa-2017-zatul-comparison%20of%20purification%20method.pdf |
| Summary: | Recombinant bromelain is a cysteine protease that can be exploited for its protease activity in food and pharmaceutical applications. The aim of this study was to compare different purification methods aqueous two-phase system (ATPS), ammonium sulphate precipitation, ion exchange, affinity, and gel filtration chromatography) for the recombinant bromelain purification from Escherichia coli BL21-A1. From the SDS-PAGE analysis, all methods produced band with molecular weight between 50 to 55 kDa. Among the methods used, the ATPS purification consisting of 13% (w/w) of PEG6000 and 11% (w/w) potassium phosphate at pH 7.0 was chosen as the best purification method. The method produced 16.39 ± 0.03% of yield, purification fold of 5.35 ± 0.11, and specific activity of 3.47 ± 0.11 unit/mg of recombinant bromelain. This proposed study can be used as a platform for large-scale downstream processing of recombinant bromelain in industry. |
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