Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain

Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain

The often-encountered problem such as protein degradation has driven various methods of cell lysis post fermentation used. In this paper we compare the use of homogenization, sonication, sonication with lysozyme and chemical lysis using B-PER reagent with lysozyme to extract the recombinant bromelai...

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Main Authors: Mohd Arshad, Zatul Iffah, Amid, Azura, Othman, Mohd Ezza Faiez
Format: Conference or Workshop Item
Language:English
Published: 2015
Subjects:
TP248.13 Biotechnology
Online Access:http://irep.iium.edu.my/44445/
http://irep.iium.edu.my/44445/1/iCAST-Zatul.pdf
id iium-44445
recordtype eprints
spelling iium-444452015-08-26T07:53:59Z http://irep.iium.edu.my/44445/ Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain Mohd Arshad, Zatul Iffah Amid, Azura Othman, Mohd Ezza Faiez TP248.13 Biotechnology The often-encountered problem such as protein degradation has driven various methods of cell lysis post fermentation used. In this paper we compare the use of homogenization, sonication, sonication with lysozyme and chemical lysis using B-PER reagent with lysozyme to extract the recombinant bromelain from E. coli BL21-AI. The sonication process is found to be the most effective in releasing recombinant bromelain without needed any pre-treatment. To succeed the cell disruption, the choice of sonication buffer also was investigated including phosphate buffer saline (PBS), PBS containing cysteine and EDTA (PBS-CE), sodium phosphate buffer containing cysteine and EDTA (EB-CE) and sodium phosphate buffer containing sodium chloride (LB). The highest specific enzyme activity was obtained when it was extracted with EB-CE buffer. Under sodium dodecyl sulphate polyacrylamide gel electrophoresis, the recombinant bromelain showed protein band at 55kDa. 2015 Conference or Workshop Item NonPeerReviewed application/pdf en http://irep.iium.edu.my/44445/1/iCAST-Zatul.pdf Mohd Arshad, Zatul Iffah and Amid, Azura and Othman, Mohd Ezza Faiez (2015) Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain. In: International Conference on Advancement in Science and Technology (iCAST 2015) , 10-12 August 2015, Impiana Resort Cherating, Pahang, Malaysia. (Unpublished)
repository_type Digital Repository
institution_category Local University
institution International Islamic University Malaysia
building IIUM Repository
collection Online Access
language English
topic TP248.13 Biotechnology
spellingShingle TP248.13 Biotechnology
Mohd Arshad, Zatul Iffah
Amid, Azura
Othman, Mohd Ezza Faiez
Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain
description The often-encountered problem such as protein degradation has driven various methods of cell lysis post fermentation used. In this paper we compare the use of homogenization, sonication, sonication with lysozyme and chemical lysis using B-PER reagent with lysozyme to extract the recombinant bromelain from E. coli BL21-AI. The sonication process is found to be the most effective in releasing recombinant bromelain without needed any pre-treatment. To succeed the cell disruption, the choice of sonication buffer also was investigated including phosphate buffer saline (PBS), PBS containing cysteine and EDTA (PBS-CE), sodium phosphate buffer containing cysteine and EDTA (EB-CE) and sodium phosphate buffer containing sodium chloride (LB). The highest specific enzyme activity was obtained when it was extracted with EB-CE buffer. Under sodium dodecyl sulphate polyacrylamide gel electrophoresis, the recombinant bromelain showed protein band at 55kDa.
format Conference or Workshop Item
author Mohd Arshad, Zatul Iffah
Amid, Azura
Othman, Mohd Ezza Faiez
author_facet Mohd Arshad, Zatul Iffah
Amid, Azura
Othman, Mohd Ezza Faiez
author_sort Mohd Arshad, Zatul Iffah
title Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain
title_short Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain
title_full Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain
title_fullStr Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain
title_full_unstemmed Comparison of cell disruption method for E. coli BL21-AI expressing recombinant bromelain
title_sort comparison of cell disruption method for e. coli bl21-ai expressing recombinant bromelain
publishDate 2015
url http://irep.iium.edu.my/44445/
http://irep.iium.edu.my/44445/1/iCAST-Zatul.pdf
first_indexed 2023-09-18T21:03:11Z
last_indexed 2023-09-18T21:03:11Z
_version_ 1777410759738785792

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