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Generic Protocol for Optimization of Heterologous Protein Production Using Automated Microbioreactor Technology
Published on: December 15, 2017
Recombinant bromelain production in Escherichia coli: process optimization in shake flask culture by response surface
Bala Muntari1, Azura Amid, Maizirwan Mel
1Bioprocess and Molecular Engineering Research Unit, Department of Biotechnology Engineering, Faculty of Engineering, International Islamic University Malaysia, P,O, Box 10, 50728, Kuala Lumpur, Malaysia. hamzah@iium.edu.my.
Abstract:
Bromelain, a cysteine protease with various therapeutic and industrial applications, was expressed in Escherichia coli, BL21-AI clone, under different cultivation conditions (post-induction temperature, L-arabinose concentration and post-induction period). The optimized conditions by response surface methodology using face centered central composite design were 0.2% (w/v) L-arabinose, 8 hr and 25°C. The analysis of variance coupled with larger value of R2 (0.989) showed that the quadratic model used for the prediction was highly significant (p < 0.05). Under the optimized conditions, the model produced bromelain activity of 9.2 U/mg while validation experiments gave bromelain activity of 9.6 ± 0.02 U/mg at 0.15% (w/v) L-arabinose, 8 hr and 27°C. This study had innovatively developed cultivation conditions for better production of recombinant bromelain in shake flask culture.
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