Feedback regulation and the intracellular protein profile of Streptomyces griseus in a cycloheximide fermentation
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Summary
Two-dimensional gel electrophoresis (2-D PAGE) was used to study the intracellular protein profile of Streptomyces griseus in relation to cycloheximide (CH) biosynthesis. Four proteins (CR1-CR4) were found to be significantly and specifically repressed by addition of the antibiotic (1 g/l at 72 h) to a producing fermentation. Synthesis of these proteins was specific to the idiophase, concurrent with CH production. Initial addition of CH to the production medium resulted in slightly lower synthesis rates of two of the proteins (CR1 and CR2), while significantly delaying the onset of synthesis of the other two (CR3 and C4). Finally, neutral polymetric resin was added to the fermentation to alleviate feedback regulation of CH synthesis, giving roughly a twofold increase in the antibiotic production rate. Production of proteins CR3 and CR4 was increased approximately tenfold immediately following resin addition, but returned to the control rate of synthesis after 24 h.
Keywords
Fermentation Electrophoresis Production Rate Streptomyces CycloheximidePreview
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References
- Dykstra KH (1989) Intracellular events occurring with on-line product removal in a cycloheximide fermentation. Ph. D. Dissertation, University of MichiganGoogle Scholar
- Dykstra KH, Wang HY (1987) Changes in the protein profile of Streptomyces griseus during a cycloheximide fermentation. Ann NY Acad Sci 506:511–522Google Scholar
- Goldman D, Merril CR, Polinsky RJ, Ebert MH (1982) Lymphocyte proteins in Huntington's disease: quantitative analysis by use of two-dimensional electrophoresis and computerized densitometry. Clin Chem 28:1021–1025Google Scholar
- Kominek LA (1975a) Cycloheximide production by Streptomyces griseus: control mechanisms of cycloheximide biosynthesis. Antimicrob Agents Chemother 7:856–860Google Scholar
- Kominek LA (1975b) Cycloheximide production by Streptomyces griseus: alleviation of end-product bnhibition by dialysis-extraction fermentation. Antimicrob Agents Chemother 7:861–863Google Scholar
- Liu CM, Hermann T, Miller PA (1977) Feedback Inhibition of an antibiotic: Aurodox (X5108). J Antibiot 30:244–251Google Scholar
- Neel JV, Rosenblum BB, Sing CF, Skolnick MM, Hanash SM, Sternberg S (1984) Adapting two-dimensional electrophoresis to the study of human germ-line mutation rates. In: Celis JE, Bravo R (ed) Two-dimensional gel electrophoresis: methods and applications. Academic Press, New York, pp 260–306Google Scholar
- Neidhardt FC, VanBogelen RA (1987) Protein regulatory gene for temperature controlled proteins in Escherichia coli. Biochem Biophys Res Commun 100:894–900Google Scholar
- Payne GF (1984) Enhanced Secondary metabolite production through on-line product removal. Ph. D. Dissertation, University of MichiganGoogle Scholar
- Payne GF, Wang HY (1988) Phosphate feeding to permit growth while maintaining secondary product synthesis. Appl Microbiol Biotechnol 27:572–576Google Scholar
- Payne GF (1984) Enhanced secondary metabolite production in situ product removal on the conversion of sugar to cycloheximide by Streptomyces griseus. Arch Microbiol 151:331–335Google Scholar
- Phillips TA (1988) Two-dimensional polyacrylamide gel electrophoresis of proteins. DNA Protein Eng Technol 1:5–9Google Scholar
- Takeshita M, Takahashi G, Okuda T (1962) Studies on Streptomyces antibiotic, cycloheximide. XIII. New spectrophotometric determination of cycloheximide. Chem Pharm Bull (Tokyo) 10:304–308Google Scholar
- Vanek Z, Puza M, Cudlin J, Dolezilova L (1964) Metabolites of Streptomyces noursei III: incorporation of 14C-carbon dioxide into cycloheximide. Biochem Biophys Res Commun 17 (5):532–535Google Scholar
- Wang HY (1983) Integrating biochemical separation and purification steps in fermentation processes. Ann NY Acad Sci 413:313–321Google Scholar