Summary
Glucose supplements to complex growth media of Escherichia coli affect the production of a recombinant model protein under the control of a temperature-sensitive expression system. The bacterial “Crabtree effect”, which occurs in the presence of glucose under aerobic conditions, not only represses the formation of citric acid cycle enzymes, but also represses the formation of the plasmid-encoded product even though the synthesis of this protein is under the control of the temperature-inducible lambda P R-promoter/cl857-repressor expression system. When the recombinant E. coli is grown at a moderate temperature (35° C) with protein hydrolysate and glucose as substrates, a biphasic growth and production pattern is observed. In the first phase, the cells grow with a high specific growth rate, utilizing glucose and forming glutamate as a byproduct. The intracellular level of recombinant protein is very low in this phase. Later, glutamate is consumed, indicating an active citric acid cycle. The degradation of glutamate is accompanied by the intracellular accumulation of high amounts of recombinant protein.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Amarasingham CR, Bernhard DD (1965) Regulation of 2-ketoglutarate dehydrogenase formation in Escherichia coli. J Biol Chem 240:3664–3668
Anderson KB, Meyenburg K von (1980) Are growth rates of Escherichia coli in batch cultures limited by respiration?. J Bacteriol 144:114–123
Brown SW, Meyer H-P, Fiechter A (1985) Continuous production of human leukocyte interferon with Escherichia coli and continuous cell lysis in a two stage chemostat. Appl Microbiol Biotechnol 23:5–9
Castellazzi M, Brachet P, Eisen H (1972) Isolation and characterization of deletion in bacteriophage lambda residing as prophage in E. coli K12. Mol Gen Genet 117:211–218
Gleiser IE, Bauer S (1981) Growth of E. coli W to high cell concentrations by oxygen level linked control of carbon source concentration. Biotechnol Bioeng 23:1015–1021
Gray CT, Wimpenny IWT, Mossmann MR (1966) Regulation of metabolism in facultative bacteria. II. Effects of aerobiosis, anaerobiosis and nutrition on the formation of Krebs cycle enzymes in Escherichia coli. Biochim Biophys Acta 117:33–41
Laemmli UK (1970) Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227:680–685
Meyer H-P, Leist C, Fiechter A (1984) Acetate formation in continuous culture of Escherichia coli K12 D1 on defined and complex media. J Biotechnol 1:355–358
Miller JH (1982) Experiments in molecular genetics. Cold Spring Harbour Laboratory, New York
Mori H, Yano T, Kobayashi T, Shimizu S (1979) High density cultivation of biomass in fed-batch system with DO-stat. J Chem Eng Jpn 12:313–319
Qureshi AG, Fohlin L, Bergström J (1984) Application of high-performance liquid chromatography to the determination of free amino acids in physiological fluids. J Chromatogr 297:91–100
Rinas U (1987) Produktbildung und Stabilität eines rekombinanten, temperatursensitiven Escherichia coli-Stammes. PhD Thesis University of Hannover, FRG
Yamada K, Kinoshita S, Tsunoda T, Aiba K (1972) The microbial production of amino acids. Wiley, New York
Zabeau M, Stanley K (1982) Enhanced expression of cro-β-galactosidase fusion proteins under the control of the P R-promoter of bacteriophage lambda. EMBO J 1:1217–1224
Zabriskie DW, Arcuri EJ (1986) Factors influencing productivity of fermentations employing recombinant microorganisms. Enzyme Microb Technol 8:706–717
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Rinas, U., Kracke-Helm, HA. & Schügerl, K. Glucose as a substrate in recombinant strain fermentation technology. Appl Microbiol Biotechnol 31, 163–167 (1989). https://doi.org/10.1007/BF00262456
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00262456