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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

Abstract

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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

References

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  1. Adrio JL, Demain AL. 2014. Microbial enzymes : tools for biotechnological process. Biomolecules 4: 117-139.
    Pubmed KoreaMed CrossRef
  2. Souza PM, Magalhaes PO. 2010. Application of microbial αamylase in industry – A review. Braz. J. Micribiol. 41: 850-861.
    Pubmed KoreaMed CrossRef
  3. Lowe ME. 2002. The triglyceride lipases of the pancreas. J. Lipid Res. 43: 2007-2016.
    Pubmed CrossRef
  4. Yee LN, Akoh CC, Phillips RS. 1995. Terpene ester synthesis by lipase catalyzed transesterification. Biotechnol. Lett. 17: 67-70.
    CrossRef
  5. Patel RN. 2013. Biocatalytic synthesis of chiral alcohols and amino acids for development of pharmaceuticals. Biomolecules 3: 741-777.
    Pubmed KoreaMed CrossRef
  6. Elmi F, Lee HT, Huang JY, Hsieh YC, Wang YL, Chen YJ, et al. 2005. Stereoselective esterase from Pseudomonas putida IFO12996 reveals alpha/beta hydrolase folds for D-beta-acetylthioisobutyric acid synthesis. J. Bacteriol. 187: 8470-8476.
    Pubmed KoreaMed CrossRef
  7. Ghati A, Paul G. 2015. Purification and characterization of a thermo-halophilic, alkali-stable and extremely benzene tolerant esterase from a thermo-halo tolerant Bacillus cereus strain AGP-03, isolated from ‘Bakreshwar’ hot spring, India. Process Biochem. 50: 771-781.
    CrossRef
  8. Kaiser P, Raina C, Parshad R, Johri S, Verma V, Andrabi KI, et al. 2006. A novel esterase form Bacillus subtillis (RRL 1789) : purification and characterization of the enzyme. Protein. Expr. Purif. 45: 262-268.
    Pubmed CrossRef
  9. Kim HK, Na HS, Park MS, Oh TK, Lee TS. 2004. Occurrence of ofloxacin ester-hydrolyzing esterase from Bacillus niacin EM001. J. Mol. Caltal B: Enzym. 27: 237-241.
    CrossRef
  10. Baeshen MN, Al-Hejin AM, Bora RS, Ahmed MM, Ramadan HA, Saini KS, et al. 2015. Production of biopharmaceuticals in E. coli :Current scenario and future perspective. J. Microbiol. Biotechnol. 25: 953-962.
    Pubmed CrossRef
  11. Djokic L, Spasic J, Jeremic S, Vasilievic B, Prodanovic O, Prodanovic R, et al. 2015. Immobilization of Escherichia coli cells expressing 4-oxalocrotonate tautomerase for improved biotransformation of β-nitrostyrene. Bioprocess Biosyst. Eng. 38:2389-2395.
    Pubmed CrossRef
  12. Zajkoska P, Rebros M, Rosenberg M. 2013. Biocatalysis with immobilized E. coli. Appl. Microbiol. Biotechnol. 97: 1441-1455.
    Pubmed CrossRef
  13. Shiloach J, Fass R. 2005. Growing E. coli to high cell density – a historical perspective on method development. Biotechnol. Adv. 23: 345-357.
    Pubmed CrossRef
  14. Nakagawa S, Oda H, Anazawa H. 1995. High cell density cultivation and high recombinant protein production of Escherichia coli strain expressing uricase. Biosci. Biotech. Biochem. 59:2263-2267.
    Pubmed CrossRef
  15. Lee SY. 1996. High cell density culture of Escherichia coli. Trends Biotechnol. 14: 98-105.
    CrossRef
  16. Luli GW, Strohl WR. 1990. Comparison of growth, acetate production, and acetate inhibition of Escherichia coli strains in batch and fed-batch fermentations. Appl. Environ. Microbiol. 56: 1004-1011.
    Pubmed KoreaMed
  17. Castan A, Enfors SO. 2000. Characteristics of DO controlled fed batch culture of Escherichia coli. Bioprocess Eng. 22: 509-515.
    CrossRef
  18. Yazdani SS, Shakri AR, Chitnis CE. 2004. A high cell density fermentation strategy to produce recombinant malarial antigen in E. coli. Biotechnol. Lett. 26: 1891-1895.
    Pubmed CrossRef
  19. Kovarova-Kovar K, Egli T. 1998. Growth kinetics of suspended microbial cells : From single-substrate-controlled growth to mixed-substrate kinetics. Microbiol. Mol. Biol. Rev. 62: 646-666.
    Pubmed KoreaMed
  20. Belo I, Mota M. 1998. Batch and fed batch cultures of E. coli TB1 at different oxygen transfer rate. Bioprocess Eng. 18: 451-455.
    CrossRef
  21. Liu YC, Chang WM, Lee CY. 1999. Effect of oxygen enrichment aeration on penicillin G acylase production in high cell density culture of recombinant E. coli. Bioprocess Eng. 21: 227-230.
    CrossRef
  22. Ukkonen K, Veijola J, Vasala A, Neubauer P. 2013. Effect of culture medium, host strain and oxygen transfer on recombinant Fab antibody fragment yield and leakage to medium in shaken E. coli cultures. Microb. Cell Fact. 12: 73.
    Pubmed KoreaMed CrossRef
  23. Eiteman MA, Altman E. 2006. Overcoming acetate in Escherichia coli recombinant protein fermentations. Trends Biotechnol. 24: 530-536.
    Pubmed CrossRef
  24. Suarez DC, Kilikian BV. 2000. Acetic acid accumulation in aerobic growth of recombinant Escherichia coli. Process Biochem. 35: 1051-1055.
    CrossRef
  25. Kweon DH, Han NS, Park KM, Seo JH. 2001. Overproduction of Phytolacca insularis protein in batch and fed-batch culture of recombinant Escherichia coli. Process Biochem. 36: 537-542.
    CrossRef
  26. Kilikian BV, Suarez ID, Liria CW, Gombert AK. 2000. Process strategies to improve heterologous protein production in Escherichia coli under lactose or IPTG induction. Process Biochem. 35: 1019-1025.
    CrossRef
  27. Yildir C. Onsan ZI, Kirdar B. 1998. Optimization of starting time and period of induction and inducer concentration in the production of the restriction enzyme EcoRI from recombinant Escherichia coli 294. Turk J. Chem. 22: 221-226.

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