Summary
Paracelsin, a hemolytic and membrane active polypeptide antibiotic of the peptaibol class which is excreted by the moldTrichderma reesei, was obtained by a simplified and isolation procedure utilziing hydrophobic adsorber resin. Investigation by13C nuclear magnetic resonance spectroscopy and circular dichroism revealed considerable helical portions in solution, and the very recently accomplished sequence determination of paracelsin allows the discussion of the results with regard to the closely related analogues, alamethicin and suzukacillin. A selective cleavage of the peptide was achieved by careful treatment with various acids, and a buffer of pH 8.25 and of high ionic strength made possible the quantitative determination of the C-terminal phenylalaninol released by means of ion-exchange chromatography. The significance of the production of paracelsin and related mycotoxins of the peptaibol class, exhibiting various kinds of biological activity, is discussed with respect to the extensive effort being made towards biotechnological applications of species, strains and cellulolytically highly active mutants of the fungusTrichoderma.
Similar content being viewed by others
References
Aydin, M., Bloos, D. H., König, W. A., Brückner, H., and Jung, G., Localization of the amide function in the sequence of peptide antibiotic trichotoxin A 40. Biomed. Mass Spectrom.9 (1982) 39–42.
Bailey, M. J., and Nevalainen, K. M. H., Induction, isolation and testing of stableTrichoderma reesei mutants with improved production of solubilizing cellulase. Enzyme Microb. Technol.3 (1981) 153–157.
Besch, H. R. Jr., Jones, L. R., Fleming, J. W., and Watanabe, A. M., Parallel unmasking of latent adenylase cyclase and (Na+, K+)-ATPase activities in sarcolemmal vesicles. J. biol. Chem.252 (1977) 7905–7908.
Bessler, W. G., Ottenbreit, B., Irmscher, G., and Jung, G., Interaction of membrane modifying peptide antibiotics fromTrichoderma viride with leukocytes. Biochem. biophys. Res. Commun.87 (1979) 99–105.
Boheim, G., Hanke, W., and Jung, G., Alamethicin pore formation: Voltage-dependent flip-flop of α-helix dipoles. Biophys. Struct. Mechs9 (1983) 181–191.
Bonnafous, J.-C., Dornand, J., and Mani, J.-C., Detergent-like effects of alamethicin on lymphocyte plasma membranes. Biochem. biophys. Res. Commun.86 (1979) 536–544.
Bosch, R., Brückner, H., Jung, G., and Winter, W., (−)-Isovaline: Confirmation of its D-(=R)-configuration by X-ray analysis of its N-chloroacetyl derivative. Tetrachedron38 (1982) 3579.
Brewer, D., Clader, F. W., MacInctyre, T. M., and Taylor, A., Ovine ill-thrift in Nova Scotia. I. The possible regulation of the rumen flora in sheep by the fungal flora of permanent pasture. J. agric. Sci., Camb.76 (1971) 465–477.
Brewer, D., Taylor, A., and Hoehn, M. M., Ovine ill-thrift in Nova Scotia. II. The production of antibiotics by fungi isolated from forest and marshland soil. J. agric. Sci., Camb.78 (1972) 257–264.
Brückner, H., Trichotoxin, A-40: Isolierung, Sequenzierung und Konformationsuntersuchungen eines membranmodifizierenden Polypeptid-Antibiotikums. Thesis. Universität Tübingen, FRG 1979.
Brückner, H., and Graf, H., Paracelsin, a peptide antibiotic containing α-aminoisobutyric acid, isolated fromTrichoderma reesei Simmons. Part A. Experientia39 (1983) 528–530.
Brückner, H., and Jung, G., Identification of N-acetyl-α-amino-isobutyric acid after selective trifluoroacetolysis of alamethicin and related peptide antibiotics. Chromatographia13 (1980) 170–174.
Brückner, H., Jung, G., Synthesis of L-Prolyl-leucyl-α-aminoisobutyryl-α-aminoisobutyryl-valinol and proof of identity with the isolated C-terminal fragment of trichotoxin A-40. Justus Liebigs Annln Chem. (1982) 1677–1699.
Brückner, H., Jung, G., Hanke, W., and Boheim, G., Structural requirements for membrane modifying activity in alamethiciin type antibiotics, in: Abstract of the Annual Meeting of the Deutsche Gesellschaft für Biophysik, Konstanz, FRG, p. 55. Eds G. Adam and G. Stark, Springer-Verlag, Berlin, Heidelberg, New York 1979.
Brückner, H., Jung, G., and Przybylski, M., Chromatographic and mass spectrometric characterization of the structures of the polypeptide antibiotics samarosporin and stilbellin and identity with emerimicin. Chromatographia17 (1983) 679–685.
Brückner, H., König, W. A., Greiner, M., and Jung, G., The sequences of the membrane-modifying peptide antibiotic trichotoxin A-40. Angew. Chem.91 (1979) 508–509; Angew. Chem. B18 (1979) 476–477.
Brückner, H., and Przybylski, M., Isolation and structural characterization of polypeptide antibiotics of the peptaibol class by highperfomance liquid chromatography with field desorption and fast atom bombardment mass spectrometry. J. Chromat.296 (1984) 263–275.
Doddrell, D. M., Pegg, D. T., and Bendall, M. R., Distortionless enhancement of NMR signals by polarization transfer. J. Magn. Res.48 (1982) 323–327.
Famey, J. P., and Whitehouse, M. W., About some possible anti-inflammatory properties of various membrane permeant agents. Agents Actions5 (1975) 133–136.
Ghose, T. K., and Sahai, V., Production of cellulases byTrichoderma reesei QM 9414 in fed-batch and continuous-flow culture with cell recycle. Biotechn. Bioengng21 (1979) 283–296.
Gisin, B. F., Kobayashi, S., Davis, D. G., and Hall, J. E., Synthesis of biologically active alamethicin, in: Peptides, Proceedings of the Fifth American Peptide Symposium, p. 215–217. Eds M. Goodman and J. Meienhofer. John Wiley and Sons, New York 1977.
Goksøyr, J., Eidså, G., Eriksen, J., and Osmundsvåg, K., A comparison of cellulases from different microorganisms, in: Symposium on enzymatic hydrolysis of cellulose, pp. 217–230. Eds M. Bailey, T. M. Enari and M. Linko. The Finnish National Fund for Research and Development (SITRA). Helsinki 1975.
Humphrey, A. E., Economics and utilization of enzymatically hydrolyzed cellulose, in: lit. pp. 437–453.
Irmscher, G., and Jung, G., Die hämolytischen Eigenschaften der membranmodifizierenden Peptidantibiotika Alamethicin, Suzukacillin und Trichotoxin. Eur. J. Biochem.80 (1977) 165–174.
Jung, G., Brückner, H., Bosch, R., Winter, W., Schaal, H., and Strähle, J., Ac-L-Ala-Aib-L-Ala-OMe: X-ray analysis of a distorted β-bend and magnetic nonequivalence of Aib-methyl groups. Justus Liebigs Annln Chem. (1983) 1096–1106.
Jung, G., Brückner, H., Oekonomopulos, R., Boheim, G., Breitmeier, E., and König, W. A., Structural requirements for pore formation in alamethicin and analogs, in: Peptides: Proceedings of the Sixth American Peptide Symposium, pp. 647–654. Eds E. Gross and J. Meienhofer, Pierce Chem. Co., Rockford, III. 1979.
Jung, G., Dubischar, N., and Leibfritz, D., Conformational changes of alamethicin induced by solvent and temperature. Eur. J. Biochem.54 (1975) 395–409.
Jung, G., König, W. A., Leibfritz, D., Ooka, T., Janko, K., and Boheim, G., Structural and membrane modifying properties of suzukacillin, a peptide antibiotic related to alamethicin. Part A. sequence and conformation. Biochim. biophys. Acta433 (1976) 164–181.
Katz, E., Suzukacillin, A.: Sequenzierung und Konformationsuntersuchungen eines membranmodifizierten Eikosapeptids. Thesis, Universität Tübinger, FRG 1983.
Kleinkauf, H., and Rindfleisch, H., Non-ribosomal biosynithesis of the cyclic octadecapeptide alamethicin. Acta microbiol. Acad. Sci. hung.22 (1975) 411–418.
Lau, A. L. Y., and Chan, S. I., Alamethicin-mediated fusion of lecithin vesicles. Proc. natl Acad. Sci. USA72 (1975) 2170–2174.
Mandels, M., Microbial sources of cellulase, in: Cellulose as a chemical and energy resource, pp. 81–105. Ed. C. R. Wilke. John Wiley and Sons, New York 1975.
Mandels, M., Hontz, L., and Nystrom, J., Enzymatic hydrolysis of waste cellulose. Biotechn. Bioengng16 (1974) 1471–1493.
Mandels, M., Weber, J., and Parizek, R., Enhanced cellulase production by a mutant ofTrichoderma viride. Appl. Microbiol.21 (1971) 152–154.
Meyer, C. E., and Reusser, F., A polypeptide antibacterial agent isolated fromTrichoderma viride. Experientia23 (1967) 85–86.
Montenecout, B. S., and Eveleigh, D. E., Preparation of mutants ofTrichoderma reesei with ehanced cellulase production. Appl. envir. Microbiol.34 (1977) 777–782.
Moo-Young, M., Moreira, A. R., and Tengerdy, R. P., Principles of solid-substrate fermentation, in: The Filamentous Fungi, vol. IV pp. 117–144. Eds J. E. Smith, D. R. Berry and B. Kristiansen. Edward and Arnold, London 1983.
Mueller, P., and Rudin, D. O., Action potentials induced in biomolecular lipid membranes. Nature217 (1986) 713–719.
Ooka, T. Shimojima, Y., Akimoto, T., Takeda, I., Senoh, S., and Abe, T., A new antibacterial peptide ‘suzukacillin’. Agric. biol. Chem.30 (1966) 700–702.
Ovchinnikov, Y. A., Kiryushkin, A. A., and Kozhevnikova, I. V., Mass-spectrometric determination of the amino acid sequence in peptides. Gen. Chem., USSR41 (1971) 2105–2116; translated from Zh. Obshch. Khim.41 (1971) 2085–2099.
Pandey, R. C., Cook, J. C. Jr, and Rinehart, K. L. Jr, Structures of the peptide antibiotics emerimicins III and IV. J. Am. chem. Soc.99 (1977) 5205–5206.
Pandey, R. C., Cook, J. C. Jr, and Rinehart, K. L. Jr, High resolution and field desorption mass spectrometry studies and revised structures of alamethicins I and II. J. Am. Chem. Soc.99 (1977) 8469–8483.
Pandey, R. C., Meng, H., Cook, J. C. Jr, and Rinehart, K. L. Jr, Structure of antiamoebin I from high resolution field desorption and gas chromatographic mass spectrometry studies. J. Am. chem. Soc.99 (1977) 5203–5205.
Payne, J. W., Jakes, R., and Hartley, B. S., The primary structure of alamethicin. Biochem. J.117 (1970) 757–766.
Pressman, B. C., Ionophorous antibiotics as models for biological transport. Fedn Proc.27 (1968) 1283–1288.
Przybylski, M., Fast atom bombardment and field desorption mass spectrometry: Comparative aspects of analytical development and bioanalytical application. Z. analyt. Chem.315 (1983) 402–421 and literature cited therein.
Przybylski, M., Manz, I., Dietrich, I., and Brückner, H., Elucidation of structure and microheterogeneity of the polypeptide antibotics paracelsin and trichotoxin A-50 by fast atom bombardment mass spectrometry in combination with selective in situ hydrolysis. Biomed. Mass Spectrom, in press (1984).
Raistrick, H., and Rudmann, P., Studies in thebiochemistry of micro-organisms. Biochem. J.63 (1956) 395–406.
Reusser, F., Biosynthesis of antibiotic U-22,324, a cyclic polypeptide. J. biol. Chem.242 (1967) 243–247.
Simmons, E. G., Classification of some cellulase-producingTrichoderma species, p. 618. Abstract of the Second International Mycological Congress, Tampa, Florida, USA (1977).
Thirumalachar, M. J., Antiamoebin, a new antiprotozoal-anthelmintic antibiotic. Part I. Production and biological studies. Hindustan antibiot. Bull.10 (1968) 287–289.
Toyama, N., Feasibility of sugar production from agricultural and urban cellulosic wastes withTrichoderma viride cellulase, in: Enzymatic conversion of cellulosicmaterials: Technology and application, pp. 207–219. Eds E. L. Gaden, Jr, M. H. Mandels, E. T. Reese and L. A. Spanol, John Wiley and Sons, New York 1976.
Toyama, N., and Ogawa, K., Utilization of cellulosic wastes byTrichoderma viride, in: Fermentation Technology Today, Proceedings of the IVth International Fermentation symposium, pp. 743–757. Society of Fermentation Techology, Japan 1972.
Toyama, N., and Ogawa, K., Sugar production from agricultural woody wastes by saccharification withTrichoderma viride cellulase, in: lit.
Note added in proof. This prediction was very recently established by the detection that the moldsGliocladium deliquescens andStilbella erythrocephala produce peptaibols, and the characterization of new components found in hypelcin20. Brückner, H., and Przybylski, M. Detection, isolation, sequence determination and biological properties of “peptaibols”, a class of membrane active polylogical properties of “peptaibols”, a class of membrane active polypeptides produced by microfungi, in: Proceedings of the 6th European Symposium on Animal, Plant and Microbial Toxins, p. 81. Eds J. Meier, K. Stocker and T. A. Freyvogel. Basel 1984; Brückner, H., and Przybylski, M., Methods for the rapid detection, isolation and sequence determination of “peptabols” and other Aib-containing peptides of fungal origin, in: Abstracts of the 15th International Symposium on Chromatography, Nürnberg, FRG, October 1–5, 1984, abstract 3PF-04; Fujita, T., Takaishi, y., Matsuura, K., Takeda, Y., Yoshioka, Y., and Brückner, H., Further investigation of peptide antibiotic, hypelcin A: Isolation structures of hypelcins A-I, A-II, and A-IV. Chem. pharm. Bull., Japan32 (1984) 2870–2873.
Author information
Authors and Affiliations
Additional information
Acknowledgment. We thank I. Ackermann for excellent and skilled technical assistance and gratefully acknowledge the help of R. Ratz for support in CD spectroscopy.
Rights and permissions
About this article
Cite this article
Brückner, H., Graf, H. & Bokel, M. Paracelsin; characterization by NMR spectroscopy and circular dichroism, and hemolytic properties of a peptaibol antibiotic from the cellulolytically active mnoldTrichoderma reesei. Part B. Experientia 40, 1189–1197 (1984). https://doi.org/10.1007/BF01946646
Published:
Issue Date:
DOI: https://doi.org/10.1007/BF01946646