Abstract
We have used the initial-rate approach to characterize changes in the glucose consumption kinetics of baculovirus-infected Spodoptera frugiperda clone 9 (Sf9) cells with the progression of the infection process. The specific glucose consumption rate (q G) of cultured baculovirus-infected Sf9 cells was measured at 4, 8, 12, 16, and 24 h postinfection (h.p.i.) in media containing 4–35 mM glucose. Higher medium glucose concentrations resulted in higher final extracellular virus and recombinant β-galactosidase yields. q G was related to the extracellular glucose concentration by means of a Michaelis-Menten relationship. The apparent Michaelis-Menten constant (K m) for glucose consumption was found not to change significantly during the progression of the infection process, and remained between 6.2 and 7.2 mM. However, the maximal specific glucose consumption rate (q Gmax) was found to rapidly increase after infection, peaking at 16 h.p.i. at a value four times that for uninfected Sf9 cells. The kinetic analysis of glucose consumption rates in baculovirus-infected Sf9 cells presented here will aid in the optimal design and operation of bioreactor systems for the large-scale production of recombinant products from the baculovirus/insect cell system.
Similar content being viewed by others
References
Volkman, L. E., Summers, M. D., and Hsieh, C. H. (1976), J. Virol. 19, 820–832.
Wyatt, G. R. and Wyatt, S. S. (1976), in Invertebrate Tissue Culture: Applications in Medicine, Biology and Agriculture, Kurstak, E. and Maramorosch, K., eds., Academic, New York, pp. 249–255.
Grace, T. D. C. (1962), Nature 195, 788,789.
Raghunand, N. and Dale, B. E. (1996), Appl. Biochem. Biotechnol. 56, 19–35.
Devaskar, S. U. and Mueckler, M. M. (1992), Pediatr. Res. 31, 1–13.
Diamond, D. L. and Carruthers, A. (1993), J. Biol. Chem. 268, 6437–6444.
Thorens, B., Sarkar, H. K., Kaback, H. R., and Lodish, H. F. (1988), Cell 55, 281–290.
Mueckler, M. (1994), Eur. J. Biochem. 219, 713–725.
Bolles, E. and Hollenberg, C. P. (1997), FEMS Microbiol. Rev. 21, 85–111.
Bisson, L. F., Coons, D. M., Kruckberg, A. L., and Lewis, D. A. (1993), Crit. Rev. Biochem. 28, 259–308.
Henderson, P. J., Roberts, P. E., Martin, G. E., Seamon, K. B., Walmsley, A. R., Rutherford, N. G., Varela, M. F., and Griffith, J. K. (1993), Biochem. Soc. Trans. 21, 1002–1006.
Wang, M.-Y. and Wang, C. (1993), FEBS Lett. 317, 241–244.
Fullbright, G., Lacy, E. R., and Bullesbach, E. E. (1997), Eur. J. Biochem. 245, 774–780.
Tsuzuki, S., Masuta, T., Furuno, M., Sakurai, S., and Iwami, M. (1997), Comp. Biochem. Physiol. 117, 409–416.
Nagata, K., Hatanaka, H., Kohda, D., and Kataoka, H., Nagasawa, H., Isogai, A., Ishizaki, H., Suzuki, A., and Inagaki, F. (1995), J. Mol. Biol. 253, 759–770.
Weber, M. J., Evans, P. K., Johnson, M. A., McNair, T. F., Nakamura, K. D., and Salter, D. W. (1984), Fed. Proc. 43, 107–112.
Flier, J. S., Mueckler, M. M., Usher, P., and Lodish, H. F. (1987), Science 235, 1492–1495.
Gray, M. A., James, M. H., Booth, J. C., and Pasternak, C. A. (1986), Arch. Virol. 87, 37–48.
Warren, A. P. and Pasternak, C. A. (1989), J. Cell. Physiol. 138, 323–328.
Wertheimer, E., Sasson, S., Cersai, E., and Ben-Neriah, Y. (1991), Proc. Natl. Acad. Sci. USA 88, 2525–2529.
Hink, W. F. (1970), Nature 226, 466,467.
Summers, M. D. and Smith, G. E. (1987), A Manual of Methods for Baculovirus Vectors and Insect Cell Culture Procedure, Bulletin No. 1555, Texas Agriculture Experiment Station, College Station, TX.
Luckow, V. A. and Summers, M. D. (1989), Virology 170, 31–39.
Reed, L. J. and Meunch, H. (1938), Am. J. Hygiene 27, 493–497.
Ferrance, J. P., Goel, A., and Ataai, M. M. (1993), Biotechnol. Bioeng. 42, 697–707.
Plagemann, P. G. W. and Richey, D. P. (1974), Biochim. Biophys. Acta 344, 263–305.
Roos, H. and Pfleger, K. (1972), Mol. Pharmacol. 8, 417–425.
Renner, E. D., Plagemann, P. G. W. and Bernlohr, R. W. (1972), J. Biol. Chem. 247, 5765–5776.
Gjedde, A. (1981), J. Neurochem. 36, 1463–1471.
Ter Kuile, B. H. and Opperdoes, F. R. (1992), J. Bacteriol. 174, 1273–1279.
Widnell, C. C., Baldwin, S. A., Davies, A., Martin, S., and Pasternak, C. A. (1990), FASEB J. 4, 1634–1637.
Pasternak, C. A., Aiyathurai, J. E. J., Makinde, V., Davies, A., Baldwin, S. A., Konieczko, E. M., and Widnell, C. C. (1991), J. Cell. Physiol. 149, 324–331.
Sviderskaya, E. V., Jazrawi, E., Baldwin, S. A., Widnell, C. C., and Pasternak, C. C. (1996), J. Memb. Biol. 149, 133–140.
Onetti, R., Baulida, J., and Bassols, A. (1997), FEBS Lett. 407, 267–270.
Van Lier, F. L. J., Van Duijnhoven, G. C. F., De Vaan, M. M. J. A. C. M., Vlak, J. M., and Tramper, J. (1994), Biotechnol. Prog. 10, 60–64.
Zhang, J., Kalogerakis, N., Behie, L. A., and Iatrou, K. (1993), Biotechnol. Bioeng. 42, 357–366.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Raghunand, N., Dale, B.E. Alteration of glucose consumption kinetics with progression of baculovirus infection in Spodoptera frugiperda cells. Appl Biochem Biotechnol 80, 231–242 (1999). https://doi.org/10.1385/ABAB:80:3:231
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1385/ABAB:80:3:231