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

Zatul Iffah, Mohd Arshad and Azura, Amid and Faridah, Yusof and Siti Zubaidah, Sulaiman and Siti Kholijah, Abdul Mudalip and Rohaida, Che Man and Shalyda, Md Shaarani (2017) Comparison of Purification Methods to Purify Recombinant Bromelain from Escherichia coli BL21-A1. Malaysian Journal of Analytical Sciences, 21 (4). pp. 958-971. ISSN 1394-2506

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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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