SpringerLink
Log in

Antifungal chitinases from Bacillus pumilus SG2: preliminary report

  • Original Paper
  • Published:
World Journal of Microbiology and Biotechnology Aims and scope Submit manuscript

Abstract

A halotolerant bacterium capable of chitinase production was previously isolated from saline soils of Gavkhooni marsh, Iran. The strain was identified as Bacillus pumilus strain SG2. This strain secretes two chitinases into the medium in response to the presence of chitin in the medium. The two chitinases, ChiS and ChiL, were active on both polymeric as well as oligosaccharide substrates. In this study, these two extracellular chitinases were purified from the supernatant of the Bacillus culture by ammonium sulfate precipitation. The optimum pH and temperature for both of these chitinases were pH 6 and 37°C, respectively. According to the conserved domain analysis, ChiS and ChiL were categorized in family 18 of the glycosylhydrolases. Three essential conserved amino acid residues (Asp-194, Asp-196 and Glu-198) in ChiS and (Asp-228, Asp-230 and Glu-232) in ChiL were found within the active site. Antifungal activities of the two purified chitinases were assessed on Rhizoctonia solani, Verticillium sp., Nigrospora sp., Stemphyllium botryosum and Bipolaris sp. demonstrating inhibition of all the tested strains. However, these chitinases did not inhibit cell wall formation of the non-chitin-containing fungi (oomycetes) Phytophthora citricola and Phytophthora capsici. This is the first investigation demonstrating antifungal activity of chitinases purified from B. pumilus SG2.

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

Access this article

Log in via an institution

Subscribe and save

Springer+ Basic
EUR 32.99 /Month
  • Get 10 units per month
  • Download Article/Chapter or Ebook
  • 1 Unit = 1 Article or 1 Chapter
  • Cancel anytime
Subscribe now

Buy Now

Price includes VAT (Brazil)

Instant access to the full article PDF.

Institutional subscriptions

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

Similar content being viewed by others

References

  • Ahmadian G, Degrassi G, Venturi V, Zeigler DR, Soudi M, Zanguinejad P (2007) Bacillus pumilus SG2 isolated from saline conditions produces and secretes two chitinases. J Appl Microbiol 103:1081–1089. doi:10.1111/j.1365-2672.2007.03340.x

    Article  CAS  Google Scholar 

  • Bacon CW, Yates IE, Hinton DM, Meredith F (2001) Biological control of Fusarium moniliforme in maize. Environ Health Perspect 2:325–332. doi:10.1016/j.foodcont.2005.12.013

    Article  Google Scholar 

  • Baek JH, Han BK, Jo DH (2001) Distribution of chitinase in rice (Oryza sativa L) seed and characterization of a hull specific chitinase. J Biochem Mol Biol 34:310–315

    CAS  Google Scholar 

  • Barboza-Corona JE, Nieto-Mazzocco E, Velazquez-Robledo R, Salcedo-Hernandez R, Bautista M, Jiménez B, Ibarra JE (2003) Cloning, sequencing and expression of the chitinase gene chiA74 from Bacillus thuringiensis. Appl Environ Microbiol 69:1023–1029. doi:10.1128/AEM.69.2.1023-1029.2003

    Article  CAS  Google Scholar 

  • Beahdy J (1974) Recent developments of antibiotic research and classification of antibiotics according to chemical structure. Adv Appl Microbiol 18:309–406. doi:10.1016/S0065-2164(08)70573-2

    Article  Google Scholar 

  • Boller T, Gehri A, Mauch F, Vogeli U (1983) Chitinase in bean leaves: induction by ethylene, purification, properties, and possible function. Planta 157:22–31. doi:10.1007/BF00394536

    Article  CAS  Google Scholar 

  • Bottone EJ, Peluso RW (2003) Production by Bacillus pumilus (MSH) of an antifungal compound that is active against Mucoraceae and Aspergillus species: preliminary report. J Medical Microbiol 52:69–74. doi:10.1099/jmm.0.04935-0

    Article  CAS  Google Scholar 

  • Chiou AL, Wu WS (2001) Isolation, identification and evaluation of bacterial antagonists against Botrytis elliptica on lily. J Phytopathol 149:319–324. doi:10.1046/j.1439-0434.2001.00627.x

    Article  Google Scholar 

  • Cordovilla P, Valdivia E, Gonzalez-Segura A, Galvez A, Martinez-Bueno M, Maqueda M (1993) Antagonistic action of the bacterium Bacillus licheniformis M-4 toward the amoeba Naegleria fowleri. J Eukaryot Microbiol 40:323–328. doi:10.1111/j.1550-7408.1993.tb04923.x

    Article  CAS  Google Scholar 

  • Gunarantna KR, Balasubramanian R (1994) Partial purification and properties of extracellular chitinase produced by Acremonium obclavatum, an antagonist to groundnut rust, Puccinia arachidis. World J Microbiol Biotechnol 10:342–345. doi:10.1007/BF00414876

    Article  Google Scholar 

  • Han BK, Moon JK, Ryu YW, Park YH, Jo DH (2000) Purification and characterization of acidic chitinase from gizzards of broiler (Gallus gallus L.). J Biochem Mol Biol 33:326–331

    CAS  Google Scholar 

  • Haran S, Schickler H, Chet I (1993) Increased constitutive chitinase activity in transformed Trichoderma harzianum. Biol Control 3:101–108. doi:10.1006/bcon.1993.1016

    Article  Google Scholar 

  • Huang CJ, Wang TK, Chung SC, Chen CY (2005) Identification of an antifungal chitinase from a potential biocontrol agent, Bacillus cereus 28–9. J Biochem Mol Biol 38:82–88

    CAS  Google Scholar 

  • Joo GJ (2005) Purification and characterization of an extracellular chitinase from the antifungal biocontrol agent Streptomyces halstedii. Biotechnol Lett 27:1483–1486. doi:10.1007/s10529-005-1315-y

    Article  CAS  Google Scholar 

  • Karuse N, Tenyo O, Kobaru S, Kamei H, Miyaki T, Konishi M, Oki T (1990) Pumilucidin, a complex of new antiviral antibiotics. Production, isolation, chemical properties, structure and biological activity. J Antibiot 43:267–280

    Google Scholar 

  • Katz E, Demain AL (1977) The peptide antibiotics of Bacillus: chemistry, biogenesis and possible functions. Bacteriol Rev 41:449–474

    CAS  Google Scholar 

  • Kim DS, Cork RJ, Weller DM (1997) Bacillus sp. L324–92 for biological control of three rot diseases of the wheat grown with reduced tillage. Phytopathology 87:551–558. doi:10.1094/PHYTO.1997.87.5.551

    Article  CAS  Google Scholar 

  • Kim SK, Kim YT, Byun HG, Park PJ, Ito H (2001) Purification and characterization of antioxidative peptide from bovine skin. J Biochem Mol Biol 34:219–224

    CAS  Google Scholar 

  • Kunz C, Ludwig A, Boller T (1992) Evaluation of the antifungal activity of the purified chitinase I from the filamentous fungus Aphanocladium album. FEMS Microbiol Lett 90:105–109. doi:10.1111/j.1574-6968.1992.tb05135.x

    Article  CAS  Google Scholar 

  • Lorito M, Woo SL, Fernandez IG, Colucci G, Harman GE, Pintor-Toro JA, Filippone E, Muccifora S, Lawrence C, Zoina A, Tuzun S, Scala F (1998) Genes from mycoparasitic fungi as a source for improving plant resistance to fungal pathogens. Proc Natl Acad Sci USA 95:7860–7865

    Article  CAS  Google Scholar 

  • Mabuchi N, Araki Y (2001) Cloning and sequencing of two genes encoding chitinases A and B from Bacillus cereus CH. Can J Microbiol 47:895–902. doi:10.1139/cjm-47-10-895

    Article  CAS  Google Scholar 

  • Marimoto K, Karita S, Kimura T, Sakka K, Hmiya K (1997) Cloning, sequencing, and expression of the gene encoding Clostridium paraputrificum chitinase ChiB and analysis of the functions of novel cadherin-like domains and a chitin-binding domain. J Bacteriol 179:7306–7314

    Google Scholar 

  • Mathivana N, Kabilan V, Murugesan K (1997) Production of chitinase by Fusarium chlamydosporum, a mycoparasite to groundnut rust, Puccinia arachidis. Indian J Exp Biol 35:890–893

    Google Scholar 

  • Oppenheim AB, Chet I (1992) Cloned chitinase in fungal pathogen control strategies. Trends Biotechnol 10:392–394. doi:10.1016/0167-7799(92)90281-Y

    Article  Google Scholar 

  • Pal KK, Tilak KV, Saxena AK, Dey R, Singh CS (2001) Suppression of maize root diseases caused by Macrophomina phaseolina, Fusarium moniliforme and Fusarium graminearum by plant growth promoting rhizobacteria. Microbiol Res 156:209–223. doi:10.1078/0944-5013-00103

    Article  CAS  Google Scholar 

  • Pettersen EF, Goddard TD, Huang CC, Couch GS, Greenblatt DM, Meng EC, Ferrin TE (2004) UCSF chimera—a visualization system for exploratory research and analysis. J Comput Chem 25:1605–1612. doi:10.1002/jcc.20084

    Article  CAS  Google Scholar 

  • Rast DM, Horsch M, Further R, Gooday GW (1991) A complex chitinolytic system in exponentially growing mycelium of Mucor rouxii: properties and function. J Gen Microbiol 137:2707–2810. doi:10.1099/00221287-137-12-2797

    Google Scholar 

  • Robbins PW, Overbye K, Pero J (1992) Cloning and high-level expression of chitinase-encoding gene of Streptomyces plicatus. Gene 111:69–76. doi:10.1016/0378-1119(92)90604-N

    Article  CAS  Google Scholar 

  • Roberts WK, Selitrennikoff CP (1998) Plant and bacterial chitinases differ in antifungal activity. J Gen Microbiol 134:169–176. doi:10.1099/00221287-134-1-169

    Google Scholar 

  • Sakai K, Yokota A, Kurokawa H, Wakayama M, Moriguchi M (1998) Purification and characterization of three thermostable endochitinases of a noble Bacillus strain, MH-1, isolated from chitin-containing compost. Appl Environ Microbiol 64:3397–3402

    CAS  Google Scholar 

  • Saksirirat W, Hoppe HH (1991) Secretion of extracellular enzymes by Verticillium psalliotae Treschow and Verticillium lecanii (Zimm.) Viegas during growth on uredospores of the soybean rust fungus in liquid cultures. J Phytopathol 131:161–173. doi:10.1111/j.1439-0434.1991.tb04741.x

    Article  CAS  Google Scholar 

  • Spizizen J (1958) Transformation of biochemically deficient strains of Bacillus subtilis by deoxyribonucleate. Proc Natl Acad Sci USA 44:1072–1078

    Article  CAS  Google Scholar 

  • Tantimavanich S, Pantuwatana S, Bhumiratana A, Panbangred W (1998) Multiple chitinase enzymes from a single gene of Bacillus licheniformis TP-1. J Ferment Bioeng 85:259–265. doi:10.1016/S0922-338X(97)85672-3

    Article  CAS  Google Scholar 

  • Thompson JD, Higgins DG, Gibson TJ (1994) CLUSTAL W: improving the sensitivity of progressive multiple sequence alignment through sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22:4673–4680. doi:10.1093/nar/22.22.4673

    Article  CAS  Google Scholar 

  • Tormo J, Lamed R, Chirino AJ, Morag E, Bayer EA, Shoham Y, Steitz TA (1996) Crystal structure of a bacterial family-III cellulose-binding domain: a general mechanism for attachment to cellulose. EMBO J 21:5739–5751

    Google Scholar 

  • Tsujibo H, Okami Y, Tanno H, Inamori Y (1993) Cloning, sequence, and expression of a chitinase gene from a marine bacterium, Alteromonas sp. strain O-7. J Bacteriol 175:176–181

    CAS  Google Scholar 

  • Ueno H, Miyashita K, Swada Y, Oba Y (1990) Purification and some properties of extracellular chitinase from Streptomyces sp. S-84. J Gen Appl Microbiol 36:377–392. doi:10.2323/jgam.36.377

    Article  CAS  Google Scholar 

  • Walker R, Powell AA, Seddon B (1998) Bacillus isolates from the spermosphere of peas and dwarf French beans with antifungal activity against Botrytis cinerea and Pythium species. J Appl Microbiol 84:791–801. doi:10.1046/j.1365-2672.1998.00411.x

    Article  CAS  Google Scholar 

  • Watanabe T, Ito Y, Yamada T, Hashimoto M, Sekine S, Tanaka H (1994) The roles of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. J Bacteriol 176:4465–4472

    CAS  Google Scholar 

  • Zeilinger S, Galhaup C, Payer K, Woo SL, Mach RL, Fekete C, Lorito M, Kubicek CP (1999) Chitinase gene expression during mycoparasitic interaction of Trichoderma harzianum with its host. Fungal Genet Biol 26:131–140. doi:10.1006/fgbi.1998.1111

    Article  CAS  Google Scholar 

Download references

Acknowledgments

The authors would like to thank the International Center for Genetics Engineering and Biotechnology (ICGEB, Italy) and National Institute of Genetic Engineering and Biotechnology (NIGEB, Iran) for providing financial support.

Author information

Authors and Affiliations

  1. Department of Molecular Genetic, National Institute of Genetic Engineering and Biotechnology, Tehran, Iran

    Seyedhadi Ghasemi, Gholamreza Ahmadian, Soheila Ghandili, Ali Dehestani & Parvin Shariati

  2. Department of Biotechnology and Plant Protection, College of Agriculture, University of Mazandaran, Babulsar, Iran

    Seyedhadi Ghasemi, Nadali Babaeian Jelodar, Heshmatollah Rahimian & Ali Dehestani

Authors
  1. Seyedhadi Ghasemi
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Gholamreza Ahmadian
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Nadali Babaeian Jelodar
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Heshmatollah Rahimian
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Soheila Ghandili
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Ali Dehestani
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Parvin Shariati
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Gholamreza Ahmadian.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Ghasemi, S., Ahmadian, G., Jelodar, N.B. et al. Antifungal chitinases from Bacillus pumilus SG2: preliminary report. World J Microbiol Biotechnol 26, 1437–1443 (2010). https://doi.org/10.1007/s11274-010-0318-6

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11274-010-0318-6

Keywords

Search

Navigation