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Microbiology and Biotechnology Letters

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Biocatalysis and Bioprocess Engineering  |  Bioprocess Engineering and Supporting Technologies

Microbiol. Biotechnol. Lett. 2017; 45(2): 149-154

https://doi.org/10.4014/mbl.1703.03002

Received: March 16, 2017; Accepted: May 17, 2017

재조합 대장균의 고농도 배양과 유도조건 최적화를 통한 Bacillus 유래 esterase의 생산

Optimization of Induction Conditions for Bacillus-derived Esterase Production by High-cell Density Fermentation of Recombinant Escherichia coli

Seung-Hoon Kang 1, Byung-Hyuk Min 1, Hong-Yeol Choi 1 and Dong-Il Kim 1*

Department of Biological Engineering, Inha University, Incheon 22212, Republic of Korea

To increase the efficiency of esterase production by Bacillus, high cell-density culture of recombinant Escherichia coli through fed batch fermentation was tested. Cells were cultured to OD600 of 76 (35.8 g/l DCW) with dissolved oxygen level controlled to least above 30% air saturation by supplying pure oxygen. Cells were cultured to an OD600 of 90 (42.4 g/l DCW) with glucose feeding controlled to at least 1 g/l. However, the cells reached stationary phase at the late stage of culture, despite glucose being supplied. Cells were cultured to an OD600 of 185 (87.3 g/l DCW) by supplying additional medium with fortified yeast extract. To increase the productivity of the recombinant protein, cell growth and esterase productivity based on induction time were evaluated. Late exponential phase induction for esterase production in fed batch fermentation resulted in maximum optical density OD600 of 190 (89 g/l DCW) and maximum esterase activity of 1745 U/l, corresponding to a 5.8-fold enhancement in esterase production, compared to the early exponential phase induction. In this study, we established fermentation methods for achieving maximum production of Bacillus-derived esterase by optimizing IPTG induction time in high-cell density culture by supplying pure oxygen and a nitrogen source.

Keywords: Esterase, high cell density fermentation, isopropyl β-D-1-thiogalactopyranoside induction, Escherichia coli

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