Optimal conditions for hepatitis B core antigen production in shaked flask fermentation


The effects of various environmental factors such as pH (5, 6, 7, 8 and 9), temperature (30, 37 and 40°C) and rotational speed (150, 200 and 250 rpm) on the growth and the hepatitis B core antigen (HBcAg) production ofEscherichia coli W3110IQ were examined in the present study. The highest growth rate is achieved at PH 7, 37°C and at a rotational speed of 250 rpm which is 0.927 h−1. The effect of pH on cell growth is more substantial compared to other parameters; it recorded a 123% different between the highest growth rate (0.927 h−1) at pH 7 and lowest growth at pH 5. The highest protein yield is achieved at pH 9, rotational speed of 250 rpm and 40°C. The yield of protein at pH 7 is 154% higher compared to the lowest yield achieved at pH 5. There is about 28% different of the protein yield for theE. coli cultivated at 250 rpm compared to that at 150 rpm which has the lowest HBcAg yield. The yield of protein at 40°C is 38% higher compared to the lowest yield achieved, at 30°C.


  1. [1]

    Tan, W. S. (2002) Inhibition of hepatitis B virus assembly with synthetic peptides derived from the viral surface and core antigens.J. Gen. Appl. Microbiol. 48: 103–107.

    Article  CAS  Google Scholar 

  2. [2]

    Crowther, R. A., N. A. Kiselev, B. Bottcher, J. A. Berriman, G. P. Borisova, V. Ose, and P. Pumpens (1994) Three-dimensional structure of hepatitis B virus core particles determined by electron cryomicroscopy.Cell 77: 943–950.

    Article  CAS  Google Scholar 

  3. [3]

    Touze, A., N. Enogat, Y. Buisson, and P. Coursaget (1999) Baculovirus expression of chimeric hepatitis B virus core particles with hepatitis E epitopes and their use in a hepatitis E Imunoassay.J. Clin. Microbiol. 37: 438–441.

    CAS  Google Scholar 

  4. [4]

    Murray, K. (1988) Application of recombinant DNA techniques in the development of viral vaccines.Vaccine 6: 164–174.

    Article  CAS  Google Scholar 

  5. [5]

    Pumpens, P. and E. Grens (2001) HBV core particles as a carrier for B cell/T cell epitopes.Intervirology 44: 98–114.

    Article  CAS  Google Scholar 

  6. [6]

    Tan, W. S., M. R. Dyson, and K. Murray (1999) Two distinct segments of the hepatitis B virus surface antigen contribute synergistically to its association with the viral core particles.J. Mol. Biol. 286: 797–808.

    Article  CAS  Google Scholar 

  7. [7]

    Brown, T. A. (1988)Gene Cloning: An Introduction. pp. 188–203, VNR International, Berkshire, UK.

    Google Scholar 

  8. [8]

    Shuler, M. L. and F. Kargi (2002)Bioprocess Engineering: Basic Concepts. 2nd ed., pp. 155–189. Prentice Hall, NJ, USA.

    Google Scholar 

  9. [9]

    Herendeen, S. L., R. A. VanBogelen, and F. C. Neidhardt (1979) Levels of major proteins ofEscherichia coli during growth at different temperatures.J. Bacteriol. 139: 185–94.

    CAS  Google Scholar 

  10. [10]

    Duffy, G., R. C. Whiting, and J. J. Sheridan (1999) The effect of a competitive microflora, pH and temperature on the growth kinetics ofEscherichia coli O157:H7.Food Microbiol. 16: 299–307.

    Article  Google Scholar 

  11. [11]

    Farewell, A. and F. C. Neidhardt (1998) Effect of temperature on in vivo protein synthetic capacity inEscherichia coli.J. Bacteriol. 180: 4704–4710.

    CAS  Google Scholar 

  12. [12]

    Duffy, G., D. C. Riordan, J. J. Sheridan, J. E. Call, R. C. Whiting, I. S. Blair, and D. A. McDowell (2000) Effect of pH on survival, thermotolerance, and verotoxin production ofEscherichia coli O157:H7 during simulated fermentation and storage.J Food Prot. 63: 12–18.

    CAS  Google Scholar 

  13. [13]

    Coleman, M. E., M. L. Tamplin, J. G. Phillips, and B. S. Marmer (2002) Influence of agitation, inoculum density, pH, and strain on the growth parameters ofEscherichia coli O157:H7-relevance to risk assessment.Intl. J. Food Microbiol. 2589: 399–415.

    Google Scholar 

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Correspondence to Beng Ti Tey.

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Tey, B.T., Yong, K.H., Ong, H.P. et al. Optimal conditions for hepatitis B core antigen production in shaked flask fermentation. Biotechnol Bioproc E 9, 374–378 (2004). https://doi.org/10.1007/BF02933060

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  • E. coli
  • HBcAg
  • fermentation
  • pH
  • temperature
  • rotational speed