Expression of biologically active, mature human granulocyte-macrophage colony stimulating factor with an E. coli secretory expression system

Human granulocyte-macrophage colony stimulating factor (HuGM-CSF) was expressed periplasmically in Escherichia coli with the secretory vector, pINIIIompA2. HuGM-CSF protein thus expressed was shown to be faithfully cleaved and biologically active. This protein, however, could not be released by osmo...

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Bibliographic Details
Published in:Current microbiology 1988-11, Vol.17 (6), p.321-332
Main Authors: GREENBERG, R, LUNDELL, D, NARULA, S. K, KASTELEIN, R, VAN KIMMENADE, A, ALROY, Y, BONITZ, S, CONDON, R, FOSSETTA, J, FROMMER, B, GEWAIN, K, KATZ, M, LEIBOWITZ, P. J
Format: Article
Language:English
Subjects:
Biological and medical sciences
Biotechnology
Escherichia coli
Fundamental and applied biological sciences. Psychology
Health. Pharmaceutical industry
Industrial applications and implications. Economical aspects
Other active biomolecules
Production of active biomolecules
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Summary:Human granulocyte-macrophage colony stimulating factor (HuGM-CSF) was expressed periplasmically in Escherichia coli with the secretory vector, pINIIIompA2. HuGM-CSF protein thus expressed was shown to be faithfully cleaved and biologically active. This protein, however, could not be released by osmotic shock and, on subcellular fractionation, co-sedimented with the outer membrane fraction. The effect of promoters, vectors, host strains, induction conditions and media formulation on expression levels was also evaluated. Some of these factors play a significant role in determining maximal achievable levels of HuGM-CSF in the secretory expression system of E. coli .
ISSN:0343-8651
1432-0991
DOI:10.1007/BF01570872