Optimization on cell disruption of E. coli BL21-AI expressing recombinant bromelain
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 nonnative host microorganism, the overall production...
| Main Authors: | , , , |
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| Format: | Article |
| Language: | English English |
| Published: |
Journal of Pure and Applied Microbiology
2014
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| Subjects: | |
| Online Access: | http://irep.iium.edu.my/38451/ http://irep.iium.edu.my/38451/ http://irep.iium.edu.my/38451/27/May_Ezza_JPAM.pdf http://irep.iium.edu.my/38451/29/38451_Optimization%20on%20cell%20disruption.SCOPUS.pdf |
| Summary: | 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 nonnative
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 other 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 |
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