ISSN: 0973-7510

E-ISSN: 2581-690X

Muhd. Ezza Faiez Othman1, Azura Amid1,2 , DzunNoraini Jimat1,2 and Mohd Jamil Aizat Jamaluddin1
1Department of Biotechnology Engineering, Faculty of Engineering, International
Islamic University Malaysia, Jalan Gombak 53100 Kuala Lumpur, Malaysia.
2Bioprocess and Biomolecular Engineering Research Unit (BPMERU), Department of
Biotechnology Engineering, Faculty of Engineering, International Islamic University
Malaysia, Jalan Gombak 53100 Kuala Lumpur, Malaysia.
J Pure Appl Microbiol. 2014;8(Spl. Edn. 1):797-802
© The Author(s). 2014
Received: 08/01/2014 | Accepted: 24/03/2014 | Published: 31/05/2014
Abstract

Recent advancements in recombinant DNA technology proved to be a promising and effective approach for more sustainable large scale productions of many therapeutic proteins. Nevertheless, since this approach involves expression of proteins in a non-native host microorganism, the overall production processes are not straight-forward due to several common challenges, such as protein degradation, especially during cell disruption stage. As the process has been subjected to both protein and host-specific, a systematic process conditioning for maximal production of recombinant protein is therefore required. In this study, a simple approach to determine optimal conditions for cell disruption using ultrasonication to isolate recombinant bromelain from E. coli BL21-AI is reported.  Suspension cells were lysed using ultrasonication which transmit sound wave in to break the cell wall. An optimized condition was obtained by response surface methodology (RSM). A three factor face-centered central composite design (FCCD) was applied to obtain the optimal process conditions consisting of amplitude, cycle and bursting period. The prediction model was further validated. Therefore, under the optimal conditions, having 20% amplitude, 0.5s cycle, and 1 minute bursting period in three times process, the specific enzyme activity of the recombinant bromelain was found to be functionally and reproducibly acceptable at 0.5270 U/mg.

Keywords

Cell lysis, Downstream, Enzyme activity, Intracellular, Recombinant protein

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© The Author(s) 2014. Open Access. This article is distributed under the terms of the Creative Commons Attribution 4.0 International License which permits unrestricted use, sharing, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made.