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

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 (Unpublished)

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Abstract

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.

Item Type:	Conference or Workshop Item (Speech/Talk)
Additional Information:	3688/44445
Uncontrolled Keywords:	cell disruption method, E. coli BL21-AI, bromelain
Subjects:	T Technology > TP Chemical technology > TP248.13 Biotechnology
Kulliyyahs/Centres/Divisions/Institutes (Can select more than one option. Press CONTROL button):	Kulliyyah of Engineering > Department of Biotechnology Engineering
Depositing User:	Prof Ts Dr Azura Amid
Date Deposited:	26 Aug 2015 15:53
Last Modified:	26 Aug 2015 15:53
URI:	http://irep.iium.edu.my/id/eprint/44445

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