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The production of recombinant beta-galactosidase inEscherichia coli in yeast extract enriched medium

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Journal of Industrial Microbiology

Summary

The productivity ofEscherichia coli biomass and recombinant beta-galactosidase was increased in Luria broth (LB) enriched with yeast extract. In flask culture under conditions of LB limitation, yeast extract suplementation gave the highest biomass (strain HB101/pRW756) stimulation per unit of component added compared with supplementation by various amounts of amino acids, vitamins, minerals, purines/pyrimidines, tryptone, casamino acids, casein peptone or gelatin peptone. The biomass production ofE. coli HB101/pRW756, XL-1 blue/puc118, XL-1 Blue FF/puc118 and TB-1/p1034 cells was stimulated in fermentor-scale experiments with additional yeast extract in LB. Total beta-galactosidase production from plasmid genes in fermentor-scale experiments was increased 105.4% in XL-1 blue/puc118 cells, 365.5% in XL-1 blue FF/puc118 cells and 421.4% in TB-1/p1034 cells by 0.5%, 1% and 1% weight per volume of additional yeast extract in LB, respectively. Depending on different strains, the increase of the enzyme production was obtained either by increased biomass, or the combination of enhanced gene expression and increased biomass. Neither the biomass nor beta-galactosidase production was stimulated in N4830/p1034 cells by the increase in yeast extract concentration in the medium.

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References

  1. Beckwith, J.R. 1964. A deletion analysis of the lac operator region inEscherichia coli. J. Mol. Biol. 8: 427–430.

    Google Scholar 

  2. Bremer, H. and P.P. Dennis. 1987. Modulation of chemical composition and other parameters of the cell by growth rate. In:Escherichia coli andSalmonella typhimurium, cellular and molecular biology (Frederick C. Neidhardt, ed.), pp. 1527–1542, American Society for Microbiology, Washington, DC.

    Google Scholar 

  3. Bullock, W.O., J.M. Fernandez and J.M. Short. 1987. XL-1 Blue: a high efficiency plasmid transforming recAEscherichia coli strain with beta-galactosidase selection. Biotechniques 5: 376–379.

    Google Scholar 

  4. Cho, SungAe, S. Scharpf, M. Franko and W.C. Vermeulen. 1985. Effect of iso-propyl-thio-β-d-galactoside concentration on the level of lac operon induction in steady stateEscherichia coli. Biochem. Biophys. Res. Commun. 128: 1268–1273.

    Google Scholar 

  5. Gottesman, M.E., S. Adhya and A. Das. 1980. Transcription antitermination by bacteriophage lambda N gene product. J. Mol. Biol. 140: 57–75.

    Google Scholar 

  6. Maniatis, T., E.F. Fritsch and J. Sambrook. 1982. TransformationEscherichia coli by plasmid DNA. In: Molecular Cloning, pp. 249–253, Cold Spring Harbor Laboratory.

  7. Rechcigl, M., Jr. 1978. Section G. In: Handbook Series in Nutrition and Food, Volume III, pp. 91–119, CRC Press, Inc., Ohio.

    Google Scholar 

  8. Reznikoff, W. and L. Gold. 1986. Response of protein levels to copy number. In: Maximizing gene expression, pp. 354–355, Butterworth Publishers, MA.

    Google Scholar 

  9. Robbins, J.W., Jr. and K. B. Taylor. 1987. Optimization ofEscherichia coli growth by controlled addition of glucose. SIM News, Vol. 36, No. 4, p. A17.

    Google Scholar 

  10. Schleif, R.F. and Pieter C. Wensink. Enzyme assays. In: Practical methods in molecular biology, pp. 43–45, Springer Verlag, New York. Heidelberg, Berlin.

  11. Tsai, L.B., M. Mann, F. Morris, C. Rotgers and D. Fenton. 1987. The effect of organic nitrogen and glucose on the production of recombinant human insulin-like growth factor in high cell densityEscherichia coli fermentation. J. Industrial Microbiol 2: 181–187.

    Google Scholar 

  12. Ullman, A., F. Jacob and J. Monod. 1967. Characterization by in vitro complementation of a peptide corresponding to an operator proximal segment of the beta-galactosidase structural gene ofEscherichia coli. J. Mol. Biol. 24: 339–343.

    Google Scholar 

  13. Weber, A.E. and Ka-Yiu San. 1987. Persistence and expression of the plasmid pBR322 inEscherichia coli K12 cultured in complex medium. Biotech. Lett. 9: 757–760.

    Google Scholar 

  14. Yanisch-Perron, C., J. Vieira and J. Messing. 1985. Improved M13 phage cloning vectors and host strains: nucleotide sequences of the M13mp18 and puc19 vectors. Gene 33: 103–119.

    Google Scholar 

  15. Zipser, D. and P. Bhavsar. 1976. Missense mutations in thelacZ gene that result in degradation of beta-galactosidase structure in protein. J. Bacteriol 127: 1538–1543.

    Google Scholar 

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Authors and Affiliations

  1. Department of Biochemistry, University of Alabama at Birmingham, 35294, Birmingham, Alabama, U.S.A.

    Xiaoli Li, J. W. Robbins Jr. & K. B. Taylor

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  1. Xiaoli Li
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  2. J. W. Robbins Jr.
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  3. K. B. Taylor
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Li, X., Robbins, J.W. & Taylor, K.B. The production of recombinant beta-galactosidase inEscherichia coli in yeast extract enriched medium. Journal of Industrial Microbiology 5, 85–93 (1990). https://doi.org/10.1007/BF01573857

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  • DOI: https://doi.org/10.1007/BF01573857

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