Abstract
Active penicillin acylase is formed only after an in vivo post-translational processing of the polypeptide precursor. Such a maturation process is rare in procaryotes. In this work, incubation under aerated conditions, of whole recombinant E. coli cells after glucose depletion and growth cessation, i.e., during the postfermentative stage, consistently resulted in 2- to 4-fold increases in penicillin acylase activity. Such results suggest that penicillin acylase maturation occurs to a high extent even during the postfermentative stage. Accordingly, the effect of different incubation conditions, during the postfermentative stage, on penicillin acylase was determined. Incubation under anaerobic conditions resulted only in a 1.27-fold increase of enzyme activity, with respect to the end of the batch culture, whereas a 3- and 4- fold increase occurred during incubation under dissolved oxygen concentrations of 100 and 43% (with respect to air sat.), respectively. Only a small negative effect, on the maturation process, was observed during incubation with acetate concentrations above 0.6 g/L. No effect of pH, in the range of 6.0 to 8.0, was observed.
Similar content being viewed by others
References
Balasingham, K., Warburton, D., Dunhill, P. and Lilly, M. D. (1972). Biochim Biophys. Acta 276: 250–256.
Bauer, S. and Shiloach, J. (1974). Biotechnol. Bioeng. 16: 933–941.
Bhattacharya, S., Gupta, V. S., Prabhune, A. A., SivaRaman, H., Debnath, M., and Ranjekar, P. K. (1993). Enzyme Microb. Technol. 15 (12): 1070–1073.
Bowden, G. and Georgiou, G. (1988). Biotechnol. Prog. 4 (2): 97 -101.
Choi, K. S., Kim, J. A., and Kang, H. S. (1992). J. Bacteriol. 174 (19): 6270–6279.
Jacob, H. E. (1970). Advanced Microbiology. Academic Press. London. pp 91–123.
Lee Y. L., and Chang, N. H. (1988). Biotechnol. Lett. 10 (11): 787–792.
Lee Y. L., and Chang, N. H. (1990). Biotechnol. Bioeng. 36: 330–337.
Nagalakshimi V., and Pai, J. S. (1994). Biotechnol. Tech. 8 (6): 431–434.
Ospina, S., Merino, E., Ramírez, O.T., and López-Munguía, A. (1995). Biotechnol. Lett. 17 (6): 615–620.
Ramírez, O.T., Zamora, R., Espinosa, G., Merino, E., Bolívar, F., and Quintero, R. (1994a). Proc. Biochem. 29: 197–206.
Ramírez, O. T., Zamora, R., Quintero, R., and López-Munguía, A. (1994b). Enzyme Microb. Technol. 16: 895–903.
Robas, N. and Branlant, C. (1994). Curr. Microbiol. 29 (5): 263–268.
Scherrer, S., Robas, N., Zouheiry, H., and Branlant, G. (1994). Appl. Microbiol. Biotechnol. 42: 85–91.
Schewale, B.J., and SivaRaman, H. (1989). Proc. Biochem. 24 (4): 145–154.
Schumacher, G., Sizmann, D., Hang, H., Buckel, P. and Beck, A. (1986). Nucl. Acids Res. 14: 5713–5727.
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
De León, A., Galindo, E. & Ramírez, O.T. A postfermentative stage improves penicillin acylase production by a recombinant E. coli . Biotechnol Lett 18, 927–932 (1996). https://doi.org/10.1007/BF00154623
Issue Date:
DOI: https://doi.org/10.1007/BF00154623