The Korean Journal of Physiology and Pharmacology

The Korean Society of Pharmacology (대한약리학회)
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Antifungal and Anticancer Activities of a Protein from the Mushroom Cordyceps militaris

  • Park, Byung-Tae (Physical Pharmacy Laboratory, College of Pharmacy, Chung-Ang University) ;
  • Na, Kwang-Heum (Physical Pharmacy Laboratory, College of Pharmacy, Chung-Ang University) ;
  • Jung, Eui-Cha (Physical Pharmacy Laboratory, College of Pharmacy, Chung-Ang University) ;
  • Park, Jae-Wan (Physical Pharmacy Laboratory, College of Pharmacy, Chung-Ang University) ;
  • Kim, Ha-Hyung (Physical Pharmacy Laboratory, College of Pharmacy, Chung-Ang University)
  • Published : 2009.02.28
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Abstract

The mushroom Cordyceps militaris has been used for a long time in eastern Asia as a nutraceutical and in traditional Chinese medicine as a treatment for cancer patients. In the present study, a cytotoxic antifungal protease was purified from the dried fruiting bodies of C. militaris using anion-exchange chromatography on a DEAE-Sepharose column. Electrophoretic analyses indicated that this protein, designated C. militaris protein(CMP), has a molecular mass of 12 kDa and a pI of 5.1. The optimum conditions for protease activity were a temperature of $37^{\circ}C$ and pH of $7.0{\sim}9.0$. The enzyme activity was specifically inhibited by the serine protease inhibitor phenylmethylsulfonyl fluoride. Amino acid composition of intact CMP and amino acid sequences of three major peptides from a tryptic digest of CMP were determined. CMP exerted strong antifungal effect against the growth of the fungus Fusarium oxysporum, and exhibited cytotoxicity against human breast and bladder cancer cells. These results indicate that C. militaris represents a source of a novel protein that might be applied in diverse biological and medicinal applications.

Keywords

References

  1. Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72: 248-254, 1976 https://doi.org/10.1016/0003-2697(76)90527-3
  2. Cunningham KG, Manson W, Spring FS, Hutchinson SA. Cordycepin, a metabolic product isolated from cultures of Cordyceps militaris (Linn.) Link. Nature 166: 949, 1950
  3. Hattori M, Isomura S, Yokoyama E, Ujita M, Hara A. Extracellular trypsin-like proteases produced by Cordyceps militaris. J Biosci Bioeng 100: 631-636, 2005 https://doi.org/10.1263/jbb.100.631
  4. Hong SY, Oh JE, Lee KH. In vitro antifungal activity and cytotoxicity of a novel membrane-active peptide. Antimicrob Agents Chemother 43: 1704-1707, 1999
  5. Joshi BN, Sainani MN, Bastawade KB, Gupta VS, Ranjekar PK. Cysteine protease inhibitor from pearl millet: a new class of antifungal protein. Biochem Biophys Res Commun 246: 382-387, 1998 https://doi.org/10.1006/bbrc.1998.8625
  6. Jung EC, Kim KD, Bae CH, Kim JC, Kim DK, Kim HH. A mushroom lectin from ascomycete Cordyceps militaris. Biochim Biophys Acta-General Subjects 1770: 833-838, 2007 https://doi.org/10.1016/j.bbagen.2007.01.005
  7. Kim BS, Oh KT, Cho DH, Kim YJ, Koo WM, Kong KH, Kim HH. A sialic acid-binding lectin from the legume Maackia fauriei: comparison with lectins from M. amurensis. Plant Science 167: 1315-1321, 2004 https://doi.org/10.1016/j.plantsci.2004.06.029
  8. Kim JS, Sapkota K, Park SE, Choi BS, Kim S, Nguyen TH, Kim CS, Choi HS, Kim MK, Chun HS, Park Y, Kim SJ. A fibrinolytic enzyme from the medicinal mushroom Cordyceps militaris. J Microbiol 44: 622-631, 2006
  9. Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227: 680-685, 1970 https://doi.org/10.1038/227680a0
  10. Li H, Matsunaga S, Fusetani N. Halicylindramides A-C, antifungal and cytotoxic depsipeptides from the marine sponge Halichondria cylindrata. J Med Chem 38: 338-343, 1995 https://doi.org/10.1021/jm00002a015
  11. Lindequist U, Niedermeyer TH, Julich WD. The pharmacological potential of mushrooms. Evid Based Complement Alternat Med 2: 285-299, 2005 https://doi.org/10.1093/ecam/neh107
  12. Moreno AB, del Pozo AM, Borja M, Segundo BS. Activity of the antifungal protein from Aspergillus giganteus against Botrytis cinerea. Phytopathology 93: 1344-1353, 2003 https://doi.org/10.1094/PHYTO.2003.93.11.1344
  13. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65: 55-63, 1983 https://doi.org/10.1016/0022-1759(83)90303-4
  14. Nagai PHK, Ng TB. Lentin, a novel and potent antifungal protein from shitake mushroom with inhibitory effects on activity of human immunodeficiency virus-1 reverse transcriptase and proliferation of leukemia cells. Life Sciences 73: 3363-3374, 2003 https://doi.org/10.1016/j.lfs.2003.06.023
  15. Nagai PHK, Zhao Z, Ng TB. Agrocybin, an antifungal peptide from the edible mushroom Agrocybe cylindracea. Peptides 26: 191-196, 2005 https://doi.org/10.1016/j.peptides.2004.09.011
  16. Ng TB. Peptides and proteins from fungi. Peptides 25: 1055-1073, 2004 https://doi.org/10.1016/j.peptides.2004.03.013
  17. St Leger RJ, Charnley AK, Cooper RM. Characterization of cuticle-degrading proteases produced by the entomopathogen Metarhizium anisopliae. Arch Biochem Biophys 253: 221-232, 1987 https://doi.org/10.1016/0003-9861(87)90655-2
  18. Vernekar JV, Ghatge MS, Deshpande VV. Alkaline protease inhibitor: a novel class of antifungal proteins against phytopathogenic fungi. Biochem Biophys Res Commun 262: 702-727, 1999 https://doi.org/10.1006/bbrc.1999.1269
  19. Watanabe N, Hattori M, Yokoyama E, Isomura S, Ujita M, Hara A. Entomogenous fungi that produce 2,6-pyridine dicarboxylic acid (dipicolinic acid). Biosci Bioeng 102: 365-368, 2006 https://doi.org/10.1263/jbb.102.365
  20. Yen YH, Li PL, Wang CL, Wang SL. An antifungal protease produced by Pseudomonas aeruginosa M-1001 with shrimp and crab shell powder as a carbon source. Enzyme Microb Technol 39: 311-317, 2006 https://doi.org/10.1016/j.enzmictec.2005.11.050
  21. Yu R, Song L, Zhao Y, Bin W, Wang L, Zhang H, Wu Y, Ye W, Yao X. Isolation and biological properties of polysaccharide CPS-1 from cultured Cordyceps militaris. Fitoterapia 75: 465-472, 2004 https://doi.org/10.1016/j.fitote.2004.04.003