Biotechnology and Bioprocess Engineering:BBE

The Korean Society for Biotechnology and Bioengineering (한국생물공학회)
권호 표지

Enhanced Production of Recombinant Protein in Escherichia coli Using Silkworm Hemolymph

  • Kim Ji Eun (School of Chemical and Biological Engineering, Seoul National University) ;
  • Kim Eun Jeong (School of Chemical and Biological Engineering, Seoul National University) ;
  • Rhee Won Jong (School of Chemical and Biological Engineering, Seoul National University) ;
  • Park Tai Hyun (School of Chemical and Biological Engineering, Seoul National University)
  • Published : 2005.08.01
Download PDF
⟨ Previous Next ⟩

Abstract

The effect of silkworm hemolymph on the expression of recombinant protein in Escherichia coli was investigated. The addition of silkworm hemolymph to the culture medium in­creased the production of recombinant $\beta$-galactosidase in E. coli. The production was dependent on the concentration of the added silkworm hemolymph, which increased 2-, 5-, and 8-fold in media supplemented with 1, 3, and $5\%$ silkworm hemolymph, respectively. To identify the effective component, the silkworm hemolymph was fractionated by gel filtration column chromatography. A fraction, with a molecular weight of about 30 K was identified as the effective component.

Keywords

References

  1. Hockney, J. (1993) Expression systems: A user's guide. Bio/Technology 11: 887-893 https://doi.org/10.1038/nbt0893-887
  2. Marino, M. H. (1989) Expression systems for heterologous protein production. BioPharm. 2: 18-33
  3. Yoon, S. K., W. K. Kang, and T. H. Park (1994) Fed-batch operation of recombinant E. coli containing trp promoter with controlled specific growth rate. Biotechnol. Bioeng. 43: 995-999 https://doi.org/10.1002/bit.260431013
  4. Yoon, S. K., S. H. Kwon, M. G. Park, W. K. Kang, and T. H. Park (1994) Optimization of recombinant Escherichia coli fed-batch fermentation for bovine somatotropin. Biotechnol. Lett. 16: 1119-1124 https://doi.org/10.1007/BF01020836
  5. Yoon, S. K, W. K. Kang, and T. H. Park (1996) Regulation of trp promoter for production of bovine somatotropin in recombinant Escherichia coli fed-batch fermentation. J. Ferment. Bioeng. 81: 153-157 https://doi.org/10.1016/0922-338X(96)87594-5
  6. Park, S. and T. H. Park (2000) Analysis of two-stage continuous operation of Escherichia coli containing bacteriophage $\lambda$ vector. Bioprocess Eng. 23: 557-563 https://doi.org/10.1007/s004499900194
  7. Ramisetti, S., H. A. Kang, S. K. Rhee, and C. H. Kim (2003) Production of recombinant hirudin in galactokinase- deficient saccharomyces cerevisiae by fed-batch fermentation with continuous glucose feeding. Biotechnol. Bioprocess Eng. 8: 183-186 https://doi.org/10.1007/BF02935894
  8. Choi, J. I. and S. Y. Lee (2004) High level production of supra molecular weight poly(3-hydroxybutyrate) by metabolically engineered Escherichia coli. Biotechnol. Bioprocess Eng. 9: 196-200 https://doi.org/10.1007/BF02942292
  9. Lin, C. S., B. Y. Chen, T. H. Park, and H. C. Lim (1998) Characterization of bacteriophage $\lambda$ $Q^{-}$ mutant for stable and efficient production of recombinant protein in Escherichia coli system. Biotechnol. Bioeng. 57: 529-535 https://doi.org/10.1002/(SICI)1097-0290(19980305)57:5<529::AID-BIT4>3.0.CO;2-I
  10. Kim, T. S. and T. H. Park (2000) Optimization of bacteriophage $\lambda$ $Q^{-}$-containing recombinant Escherichia coli fermentation process. Bioprocess Eng. 23: 187-190
  11. Yun, E. S. and T. H. Park (2000) Quantitative measurement of general odorant using electoantennogram of male silkworm moth, Bombyx mori. Biotechnol. Bioprocess Eng. 5: 150-152 https://doi.org/10.1007/BF02931888
  12. Ha, S. H., T. H. Park, and S. E. Kim (1996) Silkworm hemolymph as a substitute for fetal bovine serum in insect cell culture. Biotech. Tech. 10: 401-406
  13. Ha, S. H. and T. H. Park (1997) Efficient production of recombinant protein in Spodoptera frugiperda/AcNPV system utilizing silkworm hemolymph. Biotechnol. Lett. 19: 1087-1091 https://doi.org/10.1023/A:1018484309194
  14. Woo, S. D., W. J. Kim, H. S. Kim, J. Y. Choi, B. R. Jin, and S. K. Kang (1997) Effect of silkworm hemolymph on the expression of E. coli $\beta$-galactosidase in insect cell lines infected with recombinant baculovirus. Mol. Cell 7: 572-574
  15. Rhee, W. J., E. J. Kim, and T. H. Park (1999) Kinetic effect of silkworm hemolymph on the delayed host cell death in an insect cell-baculovirus system. Biotechnol. Prog. 15: 1028-1032 https://doi.org/10.1021/bp990093s
  16. Rhee, W. J. and T. H. Park (2000) Silkworm hemolymph inhibits baculovirus-induced insect cell apoptosis. Biochem. Biophys. Res. Commun. 271: 186-190 https://doi.org/10.1006/bbrc.2000.2592
  17. Rhee, W. J. and T. H. Park (2001) Flow cytometric analysis of the effect of silkworm hemolymph on the baculovirus- induced insect cell apoptosis. J. Microbiol. Biotechnol. 11: 853-857
  18. Rhee, W. J., E. J. Kim, and T. H. Park (2002) Silkworm hemolymph as a potent inhibitor of apoptosis in Sf9 cells. Biochem. Biophys. Res. Commun. 295: 779-783 https://doi.org/10.1016/S0006-291X(02)00746-5
  19. Choi, S. S., W. J. Rhee, and T. H. Park (2002) Inhibition of human cell apoptosis by silkworm hemolymph. Biotechnol. Prog. 18: 874-878 https://doi.org/10.1021/bp020001q
  20. Kim, E. J. and T. H. Park (2003) Anti-apoptosis engineering. Biotechnol. Bioprocess Eng. 8: 76-82 https://doi.org/10.1007/BF02940260
  21. Kim, E. J., W. J. Rhee, and T. H. Park (2001) Isolation and characterization of an apoptosis-inhibiting component from the hemolymph of Bombyx mori. Biochem. Biophys. Res. Commun. 285: 224-228 https://doi.org/10.1006/bbrc.2001.5148
  22. J. H. Miller (1972) Purification of $\beta$-Galactosidase. In: Experiments in Molecular Genetics. pp. 398-404. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY, USA
  23. U. K. Laemmli (1970) Cleavage of structural protein during the assembly of the head of bacteriophage T4. Nature 227: 680-685 https://doi.org/10.1038/227680a0