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Optimization of culture condition for the production of D-amino acid oxidase in a recombinant Escherichia coli

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Abstract

The gene encoding D-amino acid oxidase (DAAO) from Trigonopsis variabilis CBS 4095 has been cloned and expressed in Escherichia coli BL21 (DE3). Unfortunately, it was observed that the host cell was negatively affected by the expressed DAAO, resulting in a remarkable decrease in cell growth. To overcome this problem, we investigated several factors that affect cell growth rate and DAAO production such as addition time of inducer and dissolved oxygen (DO) concentration. The addition time of lactose, which was used as an inducer, and DO concentration appeared to be critical for the cell growth of E. coli BL21 (DE3)/pET-DAAO. A two-stage DO control strategy was developed, in which the DO concentration was controlled above 50% until specific stage of bacterial growth (OD600 30–40) and then downshifted to 30% by changing the agitation speed and aeration rate, and they remained at these rates until the end of fermentation. With this strategy, the maximum DAAO activity and cell growth reached 18.5 U/mL and OD600 81, respectively. By reproducing these optimized conditions in a 12-m3 fermentor, we were able to produce DAAO at a productivity of 19 U/mL with a cell growth of OD600 80.

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Authors and Affiliations

  1. School of Life Sciences and Biotechnology, Korea University, Seoul, 136-701, Korea

    Sae-Jin Kim & Chan-Wha Kim

  2. Department of Fermentation, Chong Kun Dang Bio Corporation, Ansan, 425-100, Korea

    Nag-Jong Kim & Chang-Hun Shin

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  1. Sae-Jin Kim
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  2. Nag-Jong Kim
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  4. Chan-Wha Kim
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Correspondence to Chan-Wha Kim.

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Kim, SJ., Kim, NJ., Shin, CH. et al. Optimization of culture condition for the production of D-amino acid oxidase in a recombinant Escherichia coli . Biotechnol Bioproc E 13, 144–149 (2008). https://doi.org/10.1007/s12257-008-0005-8

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  • DOI: https://doi.org/10.1007/s12257-008-0005-8

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