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Enhanced production of carboxymethylcellulase of a marine microorganism, Bacillus subtilis subsp. subtilis A-53 in a pilot-scaled bioreactor by a recombinant Escherichia coli JM109/A-53 from rice bran

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Abstract

A gene encoding the carboxymethylcellulase (CMCase) of a marine bacterium, Bacillus subtilis subsp. subtilis A-53, was cloned in Escherichia coli JMB109 and the recombinant strain was named as E. coli JMB109/A-53. The optimal conditions of rice bran, ammonium chloride, and initial pH of the medium for cell growth, extracted by Design Expert Software based on response surface methodology, were 100.0 g/l, 7.5 g/l, and 7.0, respectively, whereas those for production of CMCase were 100.0 g/l, 7.5 g/l, and 8.0. The optimal temperatures for cell growth and the production of CMCase by E. coli JM109/A-53 were found to be and 40 and 35 °C, respectively. The optimal agitation speed and aeration rate of a 7 l bioreactor for cell growth were 400 rpm and 1.5 vvm, whereas those for production of CMCase were 400 rpm and 0.5 vvm. The optimal inner pressure for cell growth was 0.06 MPa, which was the same as that for production of CMCase. The production of CMCase by E. coli JM109/A-53 under optimized conditions was 880.2 U/ml, which was 2.9 times higher than that before optimization. In this study, rice bran and ammonium chloride were developed as carbon and nitrogen source for production of CMCase by a recombinant E. coli JM109/A-53 and the productivity of E. coli JM109/A-53 was 5.9 times higher than that of B. subtilis subp. subtilis A-53.

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Acknowledgments

This paper was financially supported by Dong-A University Research Fund.

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Authors and Affiliations

  1. Department of Medical Bioscience, Graduate School of Donga-A University, Busan, 604-714, South Korea

    Eun-Jung Lee

  2. BK21 Bio-Silver Program of Dong-A University, Busan, 604-714, South Korea

    Eun-Jung Lee, Bo-Hwa Lee, Bo-Kyung Kim & Jin-Woo Lee

  3. Department of Biotechnology, Dong-A University, Busan, 604-714, South Korea

    Bo-Hwa Lee, Bo-Kyung Kim & Jin-Woo Lee

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  1. Eun-Jung Lee
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  2. Bo-Hwa Lee
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  4. Jin-Woo Lee
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Correspondence to Jin-Woo Lee.

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Lee, EJ., Lee, BH., Kim, BK. et al. Enhanced production of carboxymethylcellulase of a marine microorganism, Bacillus subtilis subsp. subtilis A-53 in a pilot-scaled bioreactor by a recombinant Escherichia coli JM109/A-53 from rice bran. Mol Biol Rep 40, 3609–3621 (2013). https://doi.org/10.1007/s11033-012-2435-9

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  • DOI: https://doi.org/10.1007/s11033-012-2435-9

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