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Recombinant bromelain production in Escherichia coli: process optimization in shake flask culture by response surface methodology.

Muntari B, Amid A, Mel M, Jami MS, Salleh HM - AMB Express (2012)

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Bioprocess 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.

No MeSH data available.


Related in: MedlinePlus

Effects of different cell concentrations on bromelain activity by BL21-AI. Tests were carried out at varying cell concentrations (OD600 nm of 0.4-0.8), while temperature of 37°C, 0.2% (w/v) L-arabinose and 4 hr post-induction period were kept constant.
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Figure 1: Effects of different cell concentrations on bromelain activity by BL21-AI. Tests were carried out at varying cell concentrations (OD600 nm of 0.4-0.8), while temperature of 37°C, 0.2% (w/v) L-arabinose and 4 hr post-induction period were kept constant.

Mentions: Induction serves as a turning point between cell growth and recombinant protein synthesis in E. coli (Xu et al. 2006). The cloning and expression system employed requires the addition of L-arabinose to trigger the transcription of foreign gene in the plasmid and initiate the translation of recombinant protein. This leads to several changes in the metabolism of host cells. The activity of recombinant bromelain at different induction time varies widely from 0.6-1.2 U/mg (Figure 1). The maximum yield of the protein was obtained when the induction was carried out at approximate OD600 nm of 0.6. It can be seen from Figure 1 that early induction has great effects on the bromelain production.


Recombinant bromelain production in Escherichia coli: process optimization in shake flask culture by response surface methodology.

Muntari B, Amid A, Mel M, Jami MS, Salleh HM - AMB Express (2012)

Effects of different cell concentrations on bromelain activity by BL21-AI. Tests were carried out at varying cell concentrations (OD600 nm of 0.4-0.8), while temperature of 37°C, 0.2% (w/v) L-arabinose and 4 hr post-induction period were kept constant.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC3293749&req=5

Figure 1: Effects of different cell concentrations on bromelain activity by BL21-AI. Tests were carried out at varying cell concentrations (OD600 nm of 0.4-0.8), while temperature of 37°C, 0.2% (w/v) L-arabinose and 4 hr post-induction period were kept constant.
Mentions: Induction serves as a turning point between cell growth and recombinant protein synthesis in E. coli (Xu et al. 2006). The cloning and expression system employed requires the addition of L-arabinose to trigger the transcription of foreign gene in the plasmid and initiate the translation of recombinant protein. This leads to several changes in the metabolism of host cells. The activity of recombinant bromelain at different induction time varies widely from 0.6-1.2 U/mg (Figure 1). The maximum yield of the protein was obtained when the induction was carried out at approximate OD600 nm of 0.6. It can be seen from Figure 1 that early induction has great effects on the bromelain production.

Bottom Line: 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.

View Article: PubMed Central - HTML - PubMed

Affiliation: Bioprocess 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.

No MeSH data available.


Related in: MedlinePlus