Abstract
Corynebacterium glutamicum effectively secretes L-glutamate when growing under biotin limitation. The secretion of glutamate was studied with respect to kinetic and energetic parameters: rate of glutamate uptake and efflux, specificity of transport, dependence of efflux on the energy state of the cell, concentration gradient of glutamate and ions, and membrane potential. By comparing these parameters when measured in biotin-limited, i.e. “producer” cells, and biotin-supplemented, i.e. “non-producer” cells, respectively, the following conclusions could be drawn: 1. The efflux of L-glutamate in C. glutamicum cannot be explained by passive permeation of this amino acid through the plasma membrane, as it has been assumed in the generally accepted model of glutamate secretion in biotin-limited cells. 2. It is unlikely that the efflux of glutamate occurs via an inversion of the glutamate uptake system. 3. Based on our results concerning the specificity and the kinetics of glutamate transport as well as the observed regulation phenomena, we conclude that secretion of glutamate in C. glutamicum occurs by a special efflux carrier system.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
Abbreviations
- dw:
-
dry weight
- OD:
-
optical density
- TPP:
-
tetraphenyl phosphonium bromide
References
Beutler HO, Michal G (1974) L-Glutamatbestimmung mit Glutamat-Dehydrogenase, Diaphorase und Tetrazoliumsalzen. In: Bergmeyer HU (ed) Methoden der enzymatischen Analyse, Band II. Verlag Chemie, Wienheim/Bergstraße, pp 1753–1759
Bunch AW, Harris RE (1986) The manipulation of micro-organisms for the production of secondary metabolites. Biotechnol Gen Engin Rev 4:117–144
Clement Y, Lanéelle G (1986) Glutamate excretion mechanism in Corynebacterium glutamicum: triggering by biotin starvation or by surfactant addition. J Gen Microbiol 132:925–929
Clement Y, Escoffier B, Trombe MC, Lanéelle G (1984) Is glutamate excreted by its uptake system in Corynebacterium glutamicum? A working hypothesis. J Gen Microbiol 130:2589–2594
Demain AL, Birnbaum J (1968) Alteration of permeability for the release of metabolites from the microbial cell. Current Topics Microbiol Immunol 46:1–25
Ebbighausen H, Weil B, Krämer R (1988) Transport of branchedchain amino acids in Corynebacterium glutamicum. Arch Microbiol 151:238–244
Huchenq A, Marquet M, Welby M, Montrozier H, Goma G, Lanéelle G (1984) Glutamate excretion triggering mechanism: A reinvestigation of the surfactant-induced modification of cell lipids. Ann Microbiol (Inst Pasteur) 135B:53–67
Kashket ER (1985) The proton motive force in bacteria: a critical assessment of methods. Ann Rev Microbiol 39:219–242
Kikuchi M, Nakao Y (1973) Relation between cellular phospholipids and the excretion of L-glutamic acid by a glycerol auxotroph of Corynebacterium alkanolyticum. Agric Biol Chem 37:515–519
Kikuchi M, Nakao Y (1986) Glutamic acid. In: Aida K, Chibata I, Nakayama K, Takinami K, Yamada H (eds) Progress in industrial microbiology, vol 24. Kodansha Ltd, Toyko. Elsevier, Amsterdam, pp 101–116
Kimura M (1963) The effect of biotin on the amino acid biosynthesis by Micrococcus glutamicus. J Gen Appl Microbiol, Tokyo 9:205–212
Kinoshita S, Tanaka K (1972) Glutamic acid. In: Yamada K, Kinoshita S, Tsunoda T, Aida K (eds) The microbial production of amino acids. Kodansha, Tokyo. John Wiley and Sons, New York, pp 263–324
Kitano K, Sugiyama Y, Kanzaki T (1972) L-Glutamate fermentation with acetic acid by an oleic acid requiring mutant. II. Inhibitory factors against the extracellular accumulation of L-glutamate. J Ferment Technol 50:182–191
Klingenberg M, Pfaff E (1967) Means of terminating reactions. In: Estabrook RW, Pullmann MR (eds) Methods in enzymology, vol X. Academic Press Inc, New York London, pp 680–684
Krämer R, Ebbighausen H, Hoischen C (1988) Transport of amino acids in Corynebacterium glutamicum. In: Quagliariello E, Palmieri F (eds) Molecular basis of biomembrane transport. Elsevier, Amsterdam, pp 33–42
Marquet M, Uribellarrea JL, Huchenq A, Lanéelle G, Goma G (1986) Glutamate excretion by Corynebacterium glutamicum: a study of glutamate accumulation during a fermentation course. Appl Microbiol Biotechnol 25:220–223
Nakao Y, Kikuchi M, Suzuki M, Doi M (1972) Microbial production of L-glutamic acid by glycerol auxotrophs. Part I. Induction of glycerol auxotrophs and production of L-glutamic acid from n-paraffins. Agric Biol Chem 36:490–496
Nunheimer TD, Birnbaum J, Ihnen ED, Demain AL (1970) Product inhibition of the fermentative formation of glutamic acid. Appl Microbiol 20:215–217
Rottenberg H (1979) The measurement of membrane potential and pH in cells, organelles, and vesicles. In: Fleischer S, Packer L (eds) Methods in enzymology, vol LV. Academic Press Inc, New York San Francisco London, pp 547–569
Saier JRM (1979) The role of the cell surface in regulating the internal environment. In: Gunsalus IC, Sokatch JR, Ornston LN (eds) The bacteria, vol VII. Academic Press Inc, New York London, pp 167–227
Shibukawa M, Ohsawa T (1966) L-Glutamic acid fermentation with molasses. Part IV. Effect of saturated-unsaturated fatty acid ratio in the cell membrane fraction on the extracellular accumulation of L-glutamate. Agric Biol Chem 30:750–758
Shiio I, Otsuka SI, Takahashi M (1962a) Effect of biotin on the bacterial formation of glutamic acid. I. Glutamate formation and cellular permeability of amino acids. J Biochem 51:56–62
Shiio I, Otsuka SI, Katsuya N (1962b) Effect of biotin on the bacterial formation of glutamic acid. II. Metabolism of glucose. J Biochem 52:108–116
Shiio I, Narui K, Yahaba N, Takahashi M (1962c) Free intracellular amino acid of a glutamate-forming bacterium, Brevibacterium flavum No 2247. Analysis and release. J Biochem 51:109–111
Shiio I, Otsuka SI, Katsuya N (1963) Cellular permeability and extracellular formation of glutamic acid in Brevibacterium flavum. J Biochem 53:333–340
Takinami K, Yoshii H, Tsuri H, Okada H (1965) Biochemical effects of fatty acid and its derivatives on glutamic acid fermentation. Part III. Biotin-tween 60 relationship in the accumulation of L-glutamic acid and the growth of Brevibacterium lactofermentum. Agric Biol Chem 29:351–359
Takinami K, Yoshii H, Yamada Y, Okada H, Kinoshita K (1968) Control of L-glutamic acid fermentation by biotin and fatty acid. Amino Acid Nucl Acid 18:120–160
Teshiba S, Furuya A (1983) Mechanisms of 5′-inosinic acid accumulation by permeability mutants of Brevibacterium ammoniagenes. III. Intracellular 5′-IMP pool and excretion mechanisms of 5′-IMP. Agric Biol Chem 47:2357–2363
Teshiba S, Furuya A (1984) Mechanisms of 5′-inosinic acid accumulation by permeability mutants of Brevibacterium ammoniagenes. IV. Excretion mechanisms of 5′-IMP. Agric Biol Chem 48:1311–1317
Zaritsky A, Kihara M, Macnab RM (1981) Measurement of membrane potential in Bacillus subtilis: A comparison of lipophilic cations, rubidium ion, and a cyanine dye as probes. J Membr Biol 63:215–231
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Hoischen, C., Krämer, R. Evidence for an efflux carrier system involved in the secretion of glutamate by Corynebacterium glutamicum . Arch. Microbiol. 151, 342–347 (1989). https://doi.org/10.1007/BF00406562
Received:
Accepted:
Issue Date:
DOI: https://doi.org/10.1007/BF00406562