Summary
The 550-day semicontinuous cultivation of Claviceps fusiformis immobilized in calcium alginate is documented. The vegetative mycelium from seed or from early-production submerged culture is the best choice for immobilization. No extracellular glucans are produced by immobilized cells. Immobilized spores give low yields of clavine alkaloids. Alginate concentrations in a range of 2%–4% do not influence yield and spectrum of alkaloids. The cytoplasm of the immobilized cells becomes condensed (after 3 days), polysaccharides disappear, and centres of lipid synthesis are formed in the cytoplasm. After 60 days the cells harbour a great number of lipid particles, mitochondria are diminishing and their cristae partly disappear, indicating a decreased respiration capacity. After 350–500 days the volume of most cells is increased many times and the cells are filled with large oval bodies of electrondense material. Chloramphenicol protects immobilized cultures against bacterial contamination.
This is a preview of subscription content, log in via an institution to check access.
Similar content being viewed by others
References
Cheng L-J, Robbers JE, Floss HG (1980) End-product regulation of ergot alkaloid formation in intact cells and protoplasts of Claviceps species, strain SD-58, J Nat Prod 43:329–339
Dickerson AG, Mantle PG, Sczyrbak CA (1970) Autolysis of extracellular glucans produced in vitro by a strain of Claviceps fusiformis. J Gen Microbiol 60:403–415
Eich E, Eichberg D, Schwartz G, Clas F, Loos M (1985) Antimicrobial activity of clavines. Arzneimittelforsch Drug Res 35(II):1760–1762
Flieger M, Křen V, Votruba J, Sajdl P (1985) Kinetic of clavine alkaloid production by the strain Claviceps purpurea 129/35. In: Tyihák H (ed) Abstracts of conference on high performance chromatographic and electrophoretic techniques in biochemistry. Siófok, pp 27–28
Glund G, Schlee D, Reinbothe H (1979) Citric acid cycle during alkaloid production in Claviceps purpurea. Biochem Physiol Pflanzen 174:660–671
Gosmann B, Rehm H-J (1986) Oxygen uptake of microorganisms entrapped in Ca-alginate. Appl Microbiol Biotechnol 23:163–176
Kim I-S, Kim S-U, Anderson JA (1981) Microsomal agroclavine hydroxylase of Claviceps species. Phytochemistry 20:2311–2314
Kopp B, Rehm H-J (1983) Alkaloid production by immobilized mycelia of Claviceps purpurea. Eur J Appl Microbiol Biotechnol 18:257–263
Kopp B, Rehm H-J (1984) Semicontinuous cultivation of immobilized Claviceps purpurea. Appl Microbiol Biotechnol 19:141–145
Křen V, Řeháček Z, Kozová J, Sajdl P, Ludvik J (1985) Production of elymoclavine by semicontinuous cultivation of immobilized cells of fungus Claviceps (in Czech). Czech Pat AO 250474, 2. 12. 1985
Křen V, Chomátová S, Pilát P, Břemek J, Řeháček Z (1986) Effect of some broad-spectrum antibiotics on the high production strain Claviceps fusiformis W1. Biotechnol Lett 8:327–332
Křen V, Kozová J, Ludvík J, Kofroňová O, Řeháček Z (1987) Physiological activity of immobilized Claviceps cells producing clavine alkaloids. Česká Mykol (in press)
Michelon LE, Kelleher WJ (1963) The spectrofotometric determination of ergot alkaloids. A modified procedure employing paradimethylaminobenzaldehyde. Lloydia 26:129–201
Pažoutová S, Řeháček Z (1981) The role of citrate on the oxidative metabolism of submerged cultures of Claviceps purpurea 129. Arch Microbiol 129:251–253
Pažoutová S, Řeháček Z (1984) Phosphate regulation of phosphatases in submerged cultures of Claviceps purpurea 129 producing clavine alkaloids. Appl Microbiol Biotechnol 20:389–392
Pažoutová S, Flieger M, Sajdl P, Řeháček Z, Taisinger J, Bass A (1981) The relationship between intensity of oxidative metabolism and predominance of agroclavine or elymoclavine in submerged Claviceps purpurea cultures. J Nat Prod 44:225–235
Řeháček Z, Sajdl P, Dutta SM, Kozová J, Pažoutová S (1980) Influence of Krebs cycle acids on clavine alkaloid production and changes in activity of Krebs and glyoxylate cycle enzymes during fermentation. Zentralbl Bakteriol II. Abt 135:643–649
Řeháček Z, Sajdl P, Kozová J, Křen V, Rylko V (1984) Industrial mutant Claviceps fusiformis W1 overproducing elymoclavine in submerged cultivation (in Czech). Czech Pat Appl PV-8832-84 5.10.1984
Spala C (1973) Genetic problems of production of ergot alkaloids in saprophytic and parasitic conditions. In: Vaněk Z, Hošťálek Z, Cudlín J (eds) Genetics of industrial microorganisms. Elsevier, Amsterdam, pp 393–403
Wurst M, Flieger M, Řeháček Z (1978) Analysis of ergot alkaloids by high performance liquid chromatography. I. Clavines and simple derivatives of lysergic acid. J Chromatogr 150:477–483
Author information
Authors and Affiliations
Rights and permissions
About this article
Cite this article
Křen, V., Ludvík, J., Kofroňová, O. et al. Physiological activity of immobilized cells of Claviceps fusiformis during long-term semicontinuous cultivation. Appl Microbiol Biotechnol 26, 219–226 (1987). https://doi.org/10.1007/BF00286312
Received:
Revised:
Issue Date:
DOI: https://doi.org/10.1007/BF00286312