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...

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Main Authors:	Mohd Arshad, Zatul Iffah, Amid, Azura, Yusof, Faridah, Sulaiman, Siti Zubaidah, Abdul Mudalip, Siti Kholijah, Che Man, Rohaida, Md Shaarani, Shalyda
Format:	Article
Language:	English English
Published:	Faculty of Science and Technology, Universiti Kebangsaan Malaysia 2017
Subjects:	TP248.13 Biotechnology
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

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spelling	iium-584562018-04-17T02:30:07Z http://irep.iium.edu.my/58456/ Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1 Mohd Arshad, Zatul Iffah Amid, Azura Yusof, Faridah Sulaiman, Siti Zubaidah Abdul Mudalip, Siti Kholijah Che Man, Rohaida Md Shaarani, Shalyda TP248.13 Biotechnology 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. Faculty of Science and Technology, Universiti Kebangsaan Malaysia 2017-08 Article PeerReviewed application/pdf en http://irep.iium.edu.my/58456/1/58456_Comparison%20of%20purification%20methods.pdf application/pdf en http://irep.iium.edu.my/58456/2/58456_Comparison%20of%20purification%20methods_SCOPUS.pdf Mohd Arshad, Zatul Iffah and Amid, Azura and Yusof, Faridah and Sulaiman, Siti Zubaidah and Abdul Mudalip, Siti Kholijah and Che Man, Rohaida and Md Shaarani, Shalyda (2017) Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1. Malaysian Journal of Analytical Sciences, 21 (4). pp. 958-971. ISSN 1394-2506 http://www.ukm.my/mjas/v21_n4/pdf/ZatulIffah_21_4_23.pdf 10.17576/mjas-2017-2104-23
repository_type	Digital Repository
institution_category	Local University
institution	International Islamic University Malaysia
building	IIUM Repository
collection	Online Access
language	English English
topic	TP248.13 Biotechnology
spellingShingle	TP248.13 Biotechnology Mohd Arshad, Zatul Iffah Amid, Azura Yusof, Faridah Sulaiman, Siti Zubaidah Abdul Mudalip, Siti Kholijah Che Man, Rohaida Md Shaarani, Shalyda Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1
description	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.
format	Article
author	Mohd Arshad, Zatul Iffah Amid, Azura Yusof, Faridah Sulaiman, Siti Zubaidah Abdul Mudalip, Siti Kholijah Che Man, Rohaida Md Shaarani, Shalyda
author_facet	Mohd Arshad, Zatul Iffah Amid, Azura Yusof, Faridah Sulaiman, Siti Zubaidah Abdul Mudalip, Siti Kholijah Che Man, Rohaida Md Shaarani, Shalyda
author_sort	Mohd Arshad, Zatul Iffah
title	Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1
title_short	Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1
title_full	Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1
title_fullStr	Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1
title_full_unstemmed	Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1
title_sort	comparison of purification methods to purify recombinant bromelain from escherichia coli bl21-a1 = perbandingan kaedah penulenan rekombinan bromelain daripada escherichia coli bl21-a1
publisher	Faculty of Science and Technology, Universiti Kebangsaan Malaysia
publishDate	2017
url	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
first_indexed	2023-09-18T21:22:40Z
last_indexed	2023-09-18T21:22:40Z
_version_	1777411985598578688

Comparison of purification methods to purify recombinant bromelain from Escherichia coli BL21-A1 = Perbandingan kaedah penulenan rekombinan bromelain daripada Escherichia coli BL21-A1

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