Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada 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, ion exchange, affinity, and ge...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English English |
| Published: |
Faculty of Science and Technology, Universiti Kebangsaan Malaysia
2017
|
| Subjects: | |
| Online Access: | http://irep.iium.edu.my/58456/ http://irep.iium.edu.my/58456/ http://irep.iium.edu.my/58456/ http://irep.iium.edu.my/58456/1/58456_Comparison%20of%20purification%20methods.pdf http://irep.iium.edu.my/58456/2/58456_Comparison%20of%20purification%20methods_SCOPUS.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.
Rekombinan bromelain adalah sisteina protease yang boleh dieksploitasi dalam aplikasi makanan dan farmaseutikal. Tujuan
kajian ini adalah untuk membandingkan kaedah penulenan yang berbeza dengan menggunakan sistem akueus dua fasa (ATPS),
pemendakan amonium sulfat, pertukaran ion, afiniti dan gel penapisan kromatografi untuk penulenan rekombinan bromelain
daripada Escherichia coli BL21-A1. Daripada analisis SDS-PAGE, semua kaedah menghasilkan jalur dengan berat molekul
antara 50 hingga 55 kDa. Antara kaedah yang digunakan, sistem akueus dua fasa (ATPS) yang terdiri daripada 13% (w/w)
PEG6000 dan 11% (w/w) kalium fosfat pada pH 7.0 telah dipilih sebagai kaedah penulenan yang terbaik. Kaedah ini mendapat
16.39 ± 0.03% hasil, kadar penulenan 5.35 ± 0.11, dan aktiviti rekombinan bromelain sebanyak 3.47 ± 0.11 unit/mg. Kajian ini
dicadangkan boleh digunakan sebagai platform untuk pemprosesan hiliran rekombinan bromelain secara besar-besaran dalam
industri. |
|---|