Journal of Microbiology and Biotechnology

The Korean Society for Microbiology and Biotechnology (한국미생물·생명공학회)
권호 표지

Induction of the T7 Promoter Using Lactose for Production of Recombinant Plasminogen Kringle 1-3 in Escherichia coli

  • Lim, Hyung-Kwon (Mogam Biotechnology Institute, Department of Agricultural Biotechnology, Seoul National University) ;
  • Lee, Shi-Uk (Mogam Biotechnolgy Institute) ;
  • Chung, Soo-Il (Mogam Biotechnolgy Institute) ;
  • Jung, Kyung-Hwan (Mogam Biotechnolgy Institute, Department of Food and Biotechnology, Chungju National University) ;
  • Seo, Jin-Ho (Department of Agricultural Biotechnology, Seoul National University)
  • Published : 2004.04.01
Download PDF
⟨ Previous Next ⟩

Abstract

A plasminogen kringle domain 1 to 3, rKl-3, was expressed in Escherichia coli under the control of T7 promoter. For the cost-effective production of rKl-3, the induction process was analyzed and optimized. Induction characteristics with lactose were analyzed in terms of induction time and inducer concentration in various culture conditions including batch and high-cell-density fed-batch cultures. In the fed-batch culture, the induction around 6 h after initiation of the DO-stat fed-batch culture resulted in the highest expression level of rKI-3 among the induction points examined. The highest demand of oxygen at this point was crucial for the maximum expression level of rKI-3. As the lactose concentration increased, the expression level also increased, though the expression level showed a plateau above a concentration of 14 mM of lactose. Lactose acted less specifically than IPTG since most of it was hydrolyzed to glucose and galactose. However, using lactose, the cell growth and the maximum expression level of rKl-3 increased by 20% and 24%, respectively, compared with those using IPTG in the fed-batch culture. The lactose seemed to be hydrolyzed by intracellular and extracellular $\beta$-galactosidase liberated by cell lysis at the same time. Residual concentration of glucose was maintained to a a limit of detection by high performance liquid chromatography, and galactose was not consumed by the host strain Escherichia coli BL2l(DE3).

Keywords

References

  1. Nat. Biotechnol. v.17 Tumor angiogenesis - new drugs on the block Brower,V. https://doi.org/10.1038/13654
  2. Nature v.407 Angiogenesis in cancer and other diseases Carmeliet,P.;R.K.Jain https://doi.org/10.1038/35025220
  3. J. Bacteriol. v.122 Phosphorylation of D-glucose in Escherichia coli mutants defective in glucosephosphotransferase, mannosephosphotransferase, and glucokinase Curtis,S.J.;W.Epstein
  4. Enzyme Microb. Technol. v.28 Large scale production of cyclohexanone monooxygenase from Escherichia coli TOP10 pQR239 Doig,S.D.;L.M.O'Sullivan;S.Patel;J.M.Ward;J.M.Woodley https://doi.org/10.1016/S0141-0229(00)00320-3
  5. J. Ind. Microbiol. v.16 Review: Optimizing inducer and culture conditions for expression of foreign proteins under the control of the lac promoter Donovan,R.S.;C.W.Robinson;B.R.Glick https://doi.org/10.1007/BF01569997
  6. Escherichia coli and Salmonella. Cellular and Molecular Biology(2nd Ed.) Glycolysis Fraenkel,D.G.;F.C.Neidhardt(et al.)(eds.)
  7. J. Biotechnol. v.60 Recombinant gene expression in Escherichia coli cultivation using lactose as inducer Gombert,A.K.;B.V.Kilikian https://doi.org/10.1016/S0168-1656(97)00185-5
  8. Int. J. Cancer. v.82 Inhibition of human malignant glioma growth in vivo by human recombinant plasminogen kringles 1-3 Joe,Y.A.;Y.K.Hong;D.S.Chung;Y.J.Yang;J.K.Kang;Y.S.Lee;S.I.Chang;W.K.You;H.Lee;S.I.Chung https://doi.org/10.1002/(SICI)1097-0215(19990827)82:5<694::AID-IJC12>3.0.CO;2-C
  9. J. Ind. Microbiol. Biotechnol. v.18 Process optimization for large-scale production of TGF-α-PE40 in recombinant Escherichia coli: Effect of medium composition and induction timing on protein expression Lee,C.;W.J.Sun;B.W.Burgress;B.H.Junker;J.Reddy;B.C.Buckland;R.L.Greasham https://doi.org/10.1038/sj.jim.2900382
  10. Arch. Biochem. Biophys. v.375 Disruption of interkringle disulfide bond of plasminogen kringle 1-3 changes the lysine binding capability of kringle 2, but not its antiangiogenic activity Lee,H.;H.K.Kim;J.H.Lee;W.K.You;S.I.Chung;S.I.Chang;M.H.Park;Y.K.Hong;Y.A.Joe https://doi.org/10.1006/abbi.1999.1675
  11. Biotechnol. Prog. v.14 Improvement of heterologous protein productivity by controlling postinduction specific growth rate in recombinant Escherichia coli under the control of the $P_L$ promoter Lim,H.K.;K.H.Jung https://doi.org/10.1021/bp980059y
  12. Appl. Microbiol. Biotechnol. v.53 Production characteristics of interferon-a using an L-arabinose promoter system in a high-cell-density culture Lim,H.K.;K.H.Jung;D.H.Park;S.I.Chung https://doi.org/10.1007/s002530050009
  13. Escherichia coli and Salmonella. Cellular and Molecular Biology(2nd Ed.) Dissimilatory pathways for sugars, polyols, and carbohydrate Lin,E.C.C.;F.C.Neidhardt(et al.)(eds.)
  14. Lett. Appl. Microbiol. v.24 Lactose-induced expression of Bacillus sp. TS-23 amylase gene in Escherichia coli regulated by a T7 promoter Lin,L.L.;W.H.Hsu https://doi.org/10.1046/j.1472-765X.1997.00146.x
  15. FEMS Microbiol. Rev. v.14 Efficient use of lactose for the lac promoter-controlled overexpression of the main antigenic protein of the foot and mouth disease virus in Escherichia coli under fed-batch fermentation conditions Neubauer,P.;K.Hofmann https://doi.org/10.1111/j.1574-6976.1994.tb00080.x
  16. Appl. Microbiol. Biotechnol. v.36 Maximizing the expression of a recombinant gene in Escherichia coli by manipulation of induction time using lactose as inducer Neubauer,P.;K.Hofmann;O.Holst;B.Mattiasson;P.Kruschke
  17. J. Microbiol. Biotechnol. v.7 Fed-batch fermentations of recombinant Escherichia coli to produce Bacillus macerans CGTase Park,Y.C.;C.S.Kim;J.I.Kim;K.H.Choi;J.H.Seo
  18. Escherichia coli and Salmonella. Cellular and Molecular Biology(2nd Ed.) Regulation of carbon utilization Saier,M.H.;T.M.Ramseier;J.Reizer;F.C.Neidhardt(et al.)(eds.)
  19. Molecular Cloning: A Laboratory Manual(2nd ed.) Recipe A.2. Sambrook,J.;E.F.Frisch;T.Maniatis
  20. Biotechnol. Lett. v.22 Synthesis of recombinant human interleukin-2 via controlled feed of lactose-complex media in fed-batch cultures of Escherichia coli BL21(DE3)[pT7-G3IL2] Saraswat,V.;J.Lee;D.Y.Kim;Y.H.Park https://doi.org/10.1023/A:1005638602146