Skip to main content
SpringerLink
Log in

Isolation and characterization of an antifungal peptide with antiproliferative activity from seeds of Phaseolus vulgaris cv. ‘Spotted Bean’

  • Biotechnologically Relevant Enzymes and Proteins
  • Published:
Applied Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A 7.3-kDa antifungal peptide with an N-terminal sequence exhibiting remarkable homology to defensins from other leguminous plants was isolated from Phaseolus vulgaris cv. ‘Spotted Bean’. The isolation procedure involved ion exchange chromatography on O-diethylaminoethyl (DEAE) cellulose, affinity chromatography on Affi-gel blue gel, ion exchange chromatography on SP-Sepharose, and gel filtration by fast protein liquid chromatography on Superdex 75. The peptide was unadsorbed on DEAE-cellulose and adsorbed on Affi-gel blue gel and SP-Sepharose. It exerted an antifungal action on Fusarium oxysporum and Mycosphaerella arachidicola. It inhibited mycelial growth in F. oxysporum with an IC50 value of 1.8 μM. It suppressed [methyl-3H]-thymidine incorporation by leukemia L1210 cells and MBL2 cells with an IC50 value of 4.0 and 9.0 μM, respectively. There was no effect on HIV-1 reverse transcriptase activity when the peptide was tested up to 0.1 mM.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  • Chu KT, Ng TB (2003) Isolation of a large thaumatin-like antifungal protein from seeds of the Kweilin chestnut Castanopsis chinensis. Biochem Biophys Res Commun 301:364–370

    Article  CAS  Google Scholar 

  • Chu KT, Xia L, Ng TB (2005) Pleurostrin, an antifungal peptide from the oyster mushroom. Peptides 26:2098–2103

    Article  CAS  Google Scholar 

  • Gao AG, Hakimi SM, Mittanck CA, Wu Y, Woerner BM, Stark DM, Stah DM, Liang JH, Rommens CMT (2000) Fungal pathogen protection in potato by expression of a plant defensin peptide. Nat Biotechnol 18:1307–1310

    Article  CAS  Google Scholar 

  • Gozia O, Ciopraga J, Bentia T, Lungu M, Zamfirescu I, Tudor R, Roseanu A, Nitu F (1993) Antifungal properties of lectin and new chitinases from potato-tubers. FEBS Lett 370:245–249

    Google Scholar 

  • Grenier J, Potvin C, Asselin A (2000) Some fungi express β-1,3-glucanases similar to thaumatin-like proteins. Mycologia 92:841–848

    CAS  Google Scholar 

  • Guo YX, Wang HX, Ng TB (2005) Isolation of trichogin, an antifungal protein from fresh fruiting bodies of the edible mushroom Tricholoma giganteum. Peptides 26:575–580

    Article  CAS  Google Scholar 

  • Huang X, Xie WJ, Gong ZZ (2000) Characterization and antifungal activity of a chitin binding protein from Ginkgo biloba. FEBS Lett 478:123–126

    Article  CAS  Google Scholar 

  • Huynh QK, Borgmeyer JR, Zobel JF (1992) Isolation and characterization of a 22-kDa protein with antifungal properties from maize seeds. Biochem Biophys Res Commun 182:1–5

    Article  CAS  Google Scholar 

  • Joshi BN, Sainani MN, Bastawade KB, Gupta VS, Ranjekar PK (1998) Cysteine protease inhibitor from pearl millet: a new class of antifungal protein. Biochem Biophys Res Commun 246:382–387

    Article  CAS  Google Scholar 

  • Laemmli UK, Favre M (1973) Maturation of head of bacteriophage-T4.1.DNA packaging events. J Mol Biol 80:573–599

    Article  Google Scholar 

  • Lam SS, Wang HX, Ng TB (1998) Purification and characterization of novel ribosome inactivating proteins, α- and β-pisavins, from seeds of the garden pea Pisum sativum. Biochem Biophys Res Commun 253:135–142

    Article  CAS  Google Scholar 

  • Lam YW, Wang HX, Ng TB (2000) A robust cysteine-deficient chitinase-like antifungal protein from inner shoots of the edible chive Allium tuberosum. Biochem Biophys Res Commun 279:74–80

    Article  CAS  Google Scholar 

  • Leah R, Tommerup H, Svendsen I, Mundy J (1991) Biochemical and molecular characterization of three barley seed proteins with antifungal properties. J Biol Chem 266:1564–1573

    CAS  PubMed  Google Scholar 

  • Lichtenstein A, Ganz T, Selsted ME, Lehrer RI (1986) In vitro tumor-cell cytolysis mediated by peptide defensins of human and rabbit granulocytes. Blood 68:1407–1410

    Article  CAS  Google Scholar 

  • Lorito M, Broadway RM, Hayes CK, Woo SL, Noviello C, Williams DL, Harman GE (1994) Proteinase inhibitors from plants as a novel class of fungicides. Mol Plant-Microb Interact 7:525–527

    Article  CAS  Google Scholar 

  • Ng TB, Wang HX (2000) Panaxagin, a new protein from Chinese ginseng possesses antifungal, antiviral, translation-inhibiting and ribosome-inactivating activities. Life Sci 68:739–749

    Article  Google Scholar 

  • Ngai PH, Ng TB (2004) Coccinin, an antifungal peptide with antiproliferative and HIV-1 reverse transcriptase inhibitory activities from large scarlet runner beans. Peptides 25:2063–2068

    Article  CAS  Google Scholar 

  • Phillippe B, Cammue BPA, Thevissen K, Hendriks M, Eggermont K, Goderis IJ, Proost P, Van Damme J, Osborn RW, Guerbette F, Kader JC, Broekaert WF (1995) A potent antimicrobial protein from onion seeds showing sequence homology to plant lipid transfer proteins. Plant Physiol 109:445–455

    Article  Google Scholar 

  • Raj PA, Dentino AR (2002) Current status of defensins and their role in innate and adaptive immunity. FEMS Microbiol Lett 206:9–18

    Article  CAS  Google Scholar 

  • Terras FRG, Torrekens S, VanLeuven F, Osborn RW, Vanderleyden J, Cammue BPA, Broekaert WF (1993) A new family of basic cysteine-rich plant antifungal proteins from Brassicaceae species. FEBS Lett 316:233–240

    Article  CAS  Google Scholar 

  • Terras FRG, Eggermont K, Kovaleva V, Raikhel NV, Osborn RW, Kester A, Rees SB, Torrekens S, VanLeuven F, Vanderleyden J (1995) Small cysteine-rich antifungal proteins from radish: their role in host defense. Plant Cell 7:573–588

    CAS  PubMed  PubMed Central  Google Scholar 

  • Vigers AJ, Roberts WK, Selitrennikoff CP (1991) A new family of plant antifungal proteins. Mol Plant-Microb Interaction 4:315–323

    Article  CAS  Google Scholar 

  • Wang HX, Ng TB (2000) Ginkbilobin, a novel antifungal protein from Ginkgo biloba seeds with sequence similarity to embryo-abundant protein. Biochem Biophys Res Commun 279:407–411

    Article  CAS  Google Scholar 

  • Wang HX, Ng TB (2001) Isolation of a novel deoxyribonuclease with antifungal activity from Asparagus officinals seeds. Biochem Biophys Res Commun 289:120–124

    Article  CAS  Google Scholar 

  • Wang H, Ng TB (2002) Isolation of cicadin, a novel and potent antifungal peptide from juvenile cicadas. Peptides 23:7–11

    Article  Google Scholar 

  • Wong JH, Ng TB (2003) Gymnin, a potent defensin-like antifungal peptides from the Yunnan bean (Gymnocladus chinensis Baill). Peptides 24:963–968

    Article  CAS  Google Scholar 

  • Wong JH, Ng TB (2005a) Sesquin, a potent defensin-like antimicrobial peptide from ground beans with inhibitory activities toward tumor cells and HIV-1 reverse transcriptase. Peptides 26:1120–1126

    Article  CAS  Google Scholar 

  • Wong JH, Ng TB (2005b) Lunatusin, a trypsin-stable antimicrobial peptide from lima beans (Phaseolus lunatus L.). Peptides 26:2086–2092

    Article  CAS  Google Scholar 

  • Wong JH, Ng TB (2005c) Vulgarinin, a broad-spectrum antifungal peptide from haricot beans (Phaseolus vulgaris). Int J Biochem Cell Biol 37:1626–1632

    Article  CAS  Google Scholar 

  • Ye XY, Ng TB (2000) Mungin, a novel cyclophilin-like antifungal protein from the mung bean. Biochem Biophys Res Commun 273:1111–1115

    Article  CAS  Google Scholar 

  • Ye XY, Wang HX, Ng TB (2000) Sativin, a novel antifungal miraculin-like protein isolated from the legumes of the sugar snap Pisum sativum var. macrocarpon. Life Sci 67:775–781

    Article  CAS  Google Scholar 

  • Ye XY, Ng TB, Tsang PWK, Wang J (2001) Isolation of a homodimeric lectin with antifungal and antiviral activities from red kidney bean (Phaseolus vulgaris). J Protein Chem 20:367–375

    Article  CAS  Google Scholar 

  • Ye XY, Ng TB (2001) A Bowman-Birk type trypsin-chymotrypsin inhibitor from broad beans. Biochem Biophys Res Commun 289:91–96

    Article  CAS  Google Scholar 

Download references

Acknowledgement

The excellent secretarial assistance of Ms. Fion Yung is appreciated.

Author information

Authors and Affiliations

  1. State Key Laboratory for Agrobiotechnology and Department of Microbiology, China Agricultural University, Beijing, China

    H. X. Wang

  2. Department of Biochemistry, Faculty of Medicine, The Chinese University of Hong Kong, Shatin, New Territories, Hong Kong, China

    T. B. Ng

Authors
  1. H. X. Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. T. B. Ng
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to T. B. Ng.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Wang, H.X., Ng, T.B. Isolation and characterization of an antifungal peptide with antiproliferative activity from seeds of Phaseolus vulgaris cv. ‘Spotted Bean’. Appl Microbiol Biotechnol 74, 125–130 (2007). https://doi.org/10.1007/s00253-006-0650-9

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00253-006-0650-9

Keywords

Search

Navigation