High level expression of recombinant BoNT/A-Hc by high cell density cultivation of Escherichia coli |
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Authors: | Kheirollah Yari Seyed Safa-Ali Fatemi Mahmood Tavallaei |
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Affiliation: | (1) Medical Biology Research Center, Kermanshah University of Medical Sciences, Kermanshah, Iran;(2) Department of Industrial and Environmental Biotechnology, National Institute of Genetic Engineering and Biotechnology (NIGEB), Shahrak-e, Pajoohesh, km 15, Tehran-Karaj Highway, P.O. Box 14965/161, Tehran, Iran;(3) Genetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran; |
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Abstract: | The carboxylic domain of the Clostridium botulinum neurotoxin heavy chain (BoNT/A-HC), which has been reported as a vaccine candidate, contains the principle protective antigenic determinants. In this study, the high level expression of the BoNT/A-Hc was achieved by high cell density cultivation of recombinant Escherichia coli in a 2-l batch stirred-tank bioreactor. In order to maximize protein expression, post-induction time and IPTG inducer concentration were optimized by the Taguchi statistical design method. Results showed that the middle of the logarithmic phase and an IPTG concentration of 1 mM presented the optimum conditions for the maximum expression of BoNT/A-HC. High cell density cultivation was subsequently carried out as an effective strategy for the high level expression of recombinant BoNT/A-Hc. Consequently, soluble BoNT/A-Hc was produced at the maximum level of 486 mg l−1, at 3 h post-induction, which was approximately 9.3 and 7.8 times higher than the levels produced by the shake flask and batch culturing methods, respectively. |
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