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  Endophytic streptomycetes from higher plants with biological activityUSPTO Application #: 20070190633Title: Endophytic streptomycetes from higher plants with biological activity Abstract: The present invention relates to isolated strains of a Streptomyces spp. which are endophytes of dicotyledonous plants and to methods for selecting such strains. The present invention also relates to compounds having biological activity selected from the group consisting of munumbicin A, munumbicin B, munumbicin C and munumbicin D, kakadumycin A, kakadumycin B, and kakadumycin C. The present invention further relates to compositions of such compounds and to methods of protecting plants against attack by a plant pathogen and methods of inhibiting bacterial growth, fungal growth, viral infection, growth of parasitic organisms, and cancer cell growth with such compositions. (end of abstract)
Agent: Klarquist Sparkman, LLP - Portland, OR, US Inventors: Gary A. Strobel, Uvidelio F. Castillo USPTO Applicaton #: 20070190633 - Class: 435253500 (USPTO) Related Patent Categories: Chemistry: Molecular Biology And Microbiology, Micro-organism, Per Se (e.g., Protozoa, Etc.); Compositions Thereof; Proces Of Propagating, Maintaining Or Preserving Micro-organisms Or Compositions Thereof; Process Of Preparing Or Isolating A Composition Containing A Micro-organism; Culture Media Therefor, Bacteria Or Actinomycetales; Media Therefor, Streptomyces The Patent Description & Claims data below is from USPTO Patent Application 20070190633. Brief Patent Description - Full Patent Description - Patent Application Claims
CROSS-REFERENCE TO RELATED APPLICATIONS [0001] This application claims the benefit of U.S. Provisional Application No. 60/369,312, filed Apr. 3, 2002, and U.S. Provisional Application No. 60/407,782, filed Sep. 3, 2002, which applications are incorporated herein by reference. BACKGROUND OF THE INVENTION [0002] 1. Field of the Invention [0003] The present invention relates to endophytic Streptomycetes from higher plants having desirable biological activities. The present invention also relates to extracts and compounds obtainable from such strains demonstrating the desirable biological activities, including the munumbicin and kakadumycin compounds, and related families of biologically active compounds, are also provided. The present invention further relates to compositions comprising the extracts and compounds as well as methods of using the compositions. [0004] 2. Description of the Related Art [0005] The bacterial order Actinomycetales includes several genera of bacteria similar to fungi in that they have a branching, filamentous structure. The branching filaments of the Actinomycetes eventually develop a network of strands called mycelium, which are similar in appearance to the mycelium of some fungi. Actinomycetes also form spores. [0006] Actinomycetes are particularly valued for the property of producing antibiotics, with the most productive genus in this group being Streptomyces. Over 50 commercially important antibiotics have been isolated from Streptomyces spp., including streptomycin, neomycin, chloramphenicol and tetracyclines. Streptomycetes are found worldwide, and are a particularly significant as members of the soil microflora. Streptomycetes are also metabolically diverse, however, and are found in a great variety of ecological environments. [0007] Actinomycetes, in general, are not reported to be endophytes on higher plants, though recently a Streptomyces sp. was reported on an annual plant--Lolium perenne (Guerny and Mantle, 1993). This lolium endophyte produces a weak antibiotic designated as methylalbonoursin, which is a diketopiperazine, condensed from leucine and phenylalanine. Streptomycetes which are used as a source of biologically active compounds, such as antibiotics, have all been isolated from soil. [0008] The development of drug resistance in human pathogenic bacteria, such as Staphylococcus, Mycobacterium, Streptococcus, Enterococcus and others, places an ever increasing importance on the search for new antibiotics, as diseases caused by such bacteria represent a clear and growing threat to world health (NIH, 2001). For instance, tuberculosis is the second leading cause of death in the world, killing approximately 2.5 million people per year. Up to 30% of the world's peoples are carriers of this pathogen (NIH, 2001). The incidence of tuberculosis is rising in the world's population, in part due to the increased incidence of patients with HIV/AIDS, but also due to the development of drug resistance in strains of M. tuberculosis (Raviglione et al, 1995; Pablosmendez et al., 1997). [0009] In addition to the problems of drug resistance in pathogenic bacteria there is also a need for more and better antimycotics, as the human population is developing more fungal infections. This is particularly an issue with HIV/AIDS patients, but also a concern with patients with organ-transplants, who must take immunosuppressive drugs to maintain continuity of the transplanted organ. In both cases, patients with these difficulties have immune systems that are weakened. Antifungal agents that are currently available, such as amphotericin B, are toxic, and often ineffective (Walsh, 1992; Walsh and Finberg, 1999; Tiphine et al., 1999). [0010] The increased incidence of parasitic protozoan infections is a further cause of concern. The most important of these is malaria caused by Plasmodium spp. that kills up to 1.5-3 million people and produces up to nearly 500 million cases per year (NIH, 2001). It is estimated that nearly 40% of the world's population is at risk of becoming infected with malaria. Global warming as well as "airport malaria" are factors contributing to the increasing spread of this disease. Another factor contributing to the increased threat of death caused by malaria is the development of drug resistance in the Plasmodium spp. populations (NIH, 2001). In some cases, treatment of malaria and other infectious diseases has been possible with the availability of antibiotics originally derived from soil-born Streptomyces spp. (Waksman, 1967; Waksman and Lechevalier 1953; and Arai, 1976). [0011] There is also a need for environmentally sound ways to grow the world's food, and new methods of controlling pests and pathogens are continuously needed in this field, as well (Overton et al., 1996). In the past, the major source of pesticidal agents came from organic synthesis. Recently, interest has increased for using more environmentally friendly methods in agricultural production, including naturally-occurring biological compounds. [0012] It is an object of the present invention to provide endophytic streptomycetes from higher plants, and extracts and compounds thereof, with biological activity. REFERENCES [0013] Arai, T. (1976). Actinomycetes: The Boundary Microorganisms. Toppan Co. Ltd, Singapore. [0014] Bacon, C. W., and White, J. F. (2000). Microbial Endophytes. Marcel Deker Inc., N.Y. [0015] Ballio, A., Bossa, F., DiGiogio, P., Ferranti, P., Paci, M., Pucci, P., Scaloni, A., Segre, A. and Strobel, G. A. (1994). Structure of the pseudomycins, new lipodepsipeptides produced by Pseudomonas syringae MSU 16H. FEBS Letters 355, 96-100. [0016] Castillo, U. F., Strobel, G. A., Ford, E. J., Hess, W. M., Porter, H., Jensen, J. B., Albert, H., Robison, R., Condron, M. A. M., Teplow, D. B., Stevens, D., and Yaver, D. (2002) Munumbicins, wide spectrum antibiotics produced by Streptomyces munumbi endophytic on Kennedia nigriscans. Microbiology 148, 2675-2685. [0017] Goodfellow, M., Williams, S. T., and Mordarski, M. (1988). Actinomycetes in Biotechnology. Academic Press, London. [0018] Isenberg, H. D. ed. (1992). Clinical Microbiology Procedures Handbook Vol. 1, Amer. Soc. Microbiol, Washington D.C. [0019] Miller, C. M., Miller, R. V., Garton-Kinney, D., Redgrave, B., Sears, J., Condron, M., Teplow, D. and Strobel, G. A. (1998). Ecomycins, unique antimycotics from Pseudomonas viridiflava. J. Applied Microbiology. 4, 937-944. [0020] NIH (2001). NIAID Global Health Research Plan for HIV/AIDS, Malaria and Tuberculosis. U.S. Department of Health and Human Services. Bethesda, Md. [0021] Overton, J. (1996). Ecologically Based Pest Management--New Solutions for a New Century Natl. Aca. Press. Washington D.C. [0022] Pablosmendez, A., Raviglione, M. C., Laszlo, A., Binkin, N., Rieder, H. L., Bustreo, F., Cohn, D. L., Lambregtsvanweezenbeek, C. S. B., Kim, S. J., Chaulet, P., and Nunn, P. (1997). Global Surveillance for antituberculosis-drug resistance. New England J. Med. 338, 1641-1649. [0023] Raviglione, M. C., Snider, D. E., and Kochi, A. (1995). Global epidemiology of tuberculosis morbidity and mortality of a worldwide epidemic. J. Amer. Med. Assoc. 273, 220 [0024] Sato, K., Shiratori, O., and Katagiri, K. (1967). The mode of action of quinoxaline antibiotics. Interaction of quinomycin A with deoxyribonucleic acid. J. Antibiot. (Tokyo) 20, 270-276. [0025] Selva, E., Beretta, G., Montanini, N., Saddler, G. S., Gastaldo, L., Ferrari, P., Lorenzetti, R., Landini, P., Ripamonti, F., Goldstein, B. P., M. Berti, L. Montanaro, and M. Denaro. (1991). Antibiotic GE2270 a: a novel inhibitor of bacterial protein synthesis. I. Isolation and characterization. J. Antibiot (Tokyo) 44, 693-701. [0026] Silverstein, R. M., Bassler, G. C., and Morrill, T. C. (1991). Spectrometric Identification or Organic Compounds. Wiley and Sons, N.Y. [0027] Singh, M. P., Petersen, P. J., Weiss, W. J., Kong, F., and Greenstein, M. (2000). Saccharomicins, novel heptadecaglycoside antibiotics produced by Saccharothrix espanaensis: antibacterial and mechanistic activities. Antimicrob. Agents Chemother. 44, 2154-2159. [0028] Strobel, G. A., Miller, R. V., Miller, C., Condron, M., Teplow, D. B., and Hess, W. M. (1999). Cryptocandin, a potent antimycotic from the endophytic fungus Cryptosporiopsis cf. quercina. Microbiol. 145, 1919-1926. [0029] Strobel, G. A., Torczynski, R., and Bollon, A. (1997). Acremonium sp.--a leucinostatin A producing endophyte of European yew (Taxus baccata). Plant Sci. 128, 97-108. [0030] Trager, W., and Jensen, J. B. (1976). Human malaria parasites in continuous culture. Science 193, 673-675. [0031] Trager, W., and F, J. B. (1978). Cultivation of malarial parasites. Nature 273, 621-622. [0032] Waksman, S. A. and Lechevalier, H. A. (1953). Actinomycetes and Their Antibiotics. Williams and Wilkins Co., Baltimore. [0033] Waksman, S. A. (1967). The Actinomycetes. Ronald Press Co. New York. [0034] Walsh, T. A., (1992). Inhibitors of .beta.-glucan synthesis. In "Emerging Targets in Antibacterial and Antifungal Chemotherapy" pp. 340, 764-373. Ed. J. A. Sutcliffe and N. H. Georgopapadakou. London: Chapman and Hall. [0035] Walsh, T. A., and Finberg, R. W. (1999). Liposomal amphoteracin B for therapy in patients with persistent fever and neutropenia. New England J. Med. 340, 764-771. [0036] Waring, M. J., and Wakelin, L. P. G. (1974). Echinomycin: a bifunctional intercalating antibiotic. Nature 252, 653-657. [0037] Young, D. H., Michelotti, E. J., Sivendell, C. S., and Krauss, N. E. (1992). Antifungal properties of taxol and various analogues. Experientia 48, 882-885. [0038] Zhang, Y. Z., Sun, X., Zeckner, D., Sachs, B., Current, W., and Chen, S. H. (2001a). 8-Amido-bearing pseudomycin B (PSB) analogue: novel antifungal agents. Bioorg. Med. Chem. Lett. 11, 123-126. [0039] Zhang, Y. Z., Sun, X., Zeckner, D., Sachs, B., Current, W., Gidda, J., Rodriguez, M., and Chen, S. H. (2001b). Synthesis and antifungal activities of novel 3-amido bearing pseudomycin analogs. Bioorg. Med. Chem. Lett. 11, 903-907. SUMMARY OF THE INVENTION [0040] The present invention relates to isolated strains of a Streptomyces spp. which are endophytes of dicotyledonous plants. In a preferred embodiment, the isolated strain is selected from the group consisting of any one of the Streptomyces spp. of NRRL 30562, NRRL 30563, NRRL 30564, NRRL 30565, NRRL 30566, and NRRL 30567. [0041] The present invention also relates to methods for selecting a strain of endophytic Streptomyces spp. having a biological activity of interest, the method comprising the steps of (a) culturing tissue from the interior region of a dicotyledonous plant on nutrient media for a time sufficient to permit colony formation by a strain of endophytic Streptomyces spp. associated with the tissue; and (b) selecting a Streptomyces spp. strain demonstrating the biological activity of interest. The present invention also relates to strains of Streptomyces spp. selected by such a method and to extracts thereof. [0042] The present invention also relates to compounds having biological activity selected from the group consisting of munumbicin A, munumbicin B, munumbicin C and munumbicin D, kakadumycin A, kakadumycin B, and kakadumycin C. The present invention further relates to compositions of such compounds. [0043] The present invention also relates to methods of protecting plants against attack by a plant pathogen and methods of inhibiting bacterial growth, fungal growth, viral infection, growth of parasitic organisms, and cancer cell growth. BRIEF DESCRIPTION OF THE DRAWINGS [0044] FIG. 1 shows the snakevine plant growing wild in the Northern Territory of Australia. [0045] FIG. 2 shows a scanning electron micrograph of Streptomyces munumbi grown on gamma-irradiated carnation leaves. The bar equals 1 micron. Continue reading... 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