Comparison of Purification Methods to Purify Recombinant Bromelain from Escherichia coli BL21-A1

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Abstract

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.

Item Type: Article
Uncontrolled Keywords: recombinant bromelain, purification, chromatography, aqueous two-phase system
Subjects: T Technology > TP Chemical technology
Faculty/Division: Faculty of Chemical & Natural Resources Engineering
Depositing User: Noorul Farina Arifin
Date Deposited: 09 Jan 2018 02:26
Last Modified: 09 Jan 2018 02:26
URI: http://umpir.ump.edu.my/id/eprint/19928
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