• P. Velusamy Department of Biotechnology, School of Bioengineering, SRM University, Chennai, Tamil Nadu, India
  • J. Das Department of Biotechnology, School of Bioengineering, SRM University, Chennai, Tamil Nadu, India


Fusarium oxysporum, B subtilisJD-09, chitinase, antifungal activity


Objective: Chitinase plays an important role against fungal pathogen and is a hopeful solution against fungal diseases. The main of this research is to screen a chitin degrading strain which can significantly used as biocontrol agent against Fusarium oxysporum.

Methods: In the present study, chitinase producing bacteria were isolated from soil sample of crop field and identified based on biochemical observation and 16S rRNA analysis. The molecular weight of the enzyme was determined by zymogram. The purified enzyme from identified strain JD-09 was tested for antagonistic activity against Fusarium oxysporum.

Results: Strain JD-09 was screened and identified as Bacillus subtilis. The strain exhibited a maximum chitinase production of 1.33 U/mL in colloidal chitin broth after 3 days of cultivation at 28 °C. The molecular weight of the chitinase was estimated to be 57 kDa, 39 kDa and 30.7 kDa. Hydrolysis products of the fungal cell wall by the purified enzymes of B. subtilis JD-09 were analyzed by high-pressure liquid chromatography (HPLC) and identified as oligosaccharides.

Conclusion: It can be interfered that Bacillus subtilis JD-09 may be an optimal candidate for use as an antifungal agent of Fusarium wilt in crops. The purified chitinase can be directly applied for suppressing growth of living fungal hyphae.


Download data is not yet available.


Simmons KJ, Chopra I, Fishwick CWG. Structure-based discovery of antibacterial drugs. Nature reviews Microbiology 2010;8(7):501-10.

Williams PG. Panning for chemical gold:marine bacteria as a source of new therapeutics. Trends Biotechnol 2009;27(1):45-52.

Validov S, Mavrodi O, De La Fuente L, Boronin A, Weller D, Thomashow L, et al. Antagonistic activity among 2,4-diacetylphloroglucinol-producing fluorescent Pseudomonas spp. FEMS Microbiol Lett 2005;242(2):249-56.

Clardy J, Fischbach MA, Walsh CT. New antibiotics from bacterial natural products. Nat Biotechnol 2006;24(12):1541-50.

Budzikiewicz H, Fems. Secondary metabolites from fluorescent pseudomonads. Rev 1993;104:209-28.

Ramette A, Moënne-Loccoz Y, Défago G. Prevalence of fluorescent pseudomonads producing antifungal phloroglucinols and/or hydrogen cyanide in soils naturally suppressive or conducive to tobacco black root rot. FEMS Microbiol Ecol 2003;44(1):35-43.

Ayyadurai N, Naik PR, Sakthivel N. Functional characterization of antagonistic fluorescent pseudomonads associated with rhizospheric soil of rice (Oryza sativa L.). J Microbiol Biotechnol 2007;17(6):919-27.

Katz E, Demain AL. The peptide antibiotics of Bacillus:chemistry, biogenesis, and possible functions. Bacteriol Rev 1977;41(2):449-74.

Zuber, P., Nakano, M., Marshiel, M.A. Peptide antibiotics. In:Sonensheim AL, Hoch JA, Losick R. Bacillus subtilis and other Gram-positive bacteria:biochemistry, physiology, and molecular genetics. Washington, DC:American Society of Microbiology. 1993;897-916.

Yu GY, Sinclair JB, Hartman BL, Bertagnolli BL. Production of iturin A by Bacillus amyloliquefaciens suppressing Rhizoctonia solani. Soil Biol Biochem 2002;34:955-63.

Pinchuk IV, Bressollier P, Sorokulova IB, Verneuil B, Urdaci MC. Amicoumacin antibiotic production and genetic diversity of Bacillus subtilis strains isolated from different habitats. Res Microbiol 2002;153(5):269-76.

Tabernero C, Coll PM, Fernández-Abalos JM, Pérez P, Santamaría RI. Cloning and DNA sequencing of bgaA, a gene encoding an endo-beta-1,3-1,4-glucanase, from an alkalophilic Bacillus strain (N137). Appl Environ Microbiol 1994;60(4):1213-20.

Guangjun Nie, G., Yue, W., Fang, H., Jiang, Y., Shen, Y. Screening of chitinase producing strain and effect of its metabolite on Magnesium Ammonium Phosphate crystals fractal. J. Pure Appl. Microbio. 2008;2:319-24.

Chernin L, Ismailov Z, Haran S, Chet I. Chitinolytic Enterobacter agglomerans Antagonistic to Fungal Plant Pathogens. Appl Environ Microbiol 1995;61(5):1720-6.

Carlisle MJ, Watkinson SC. The Fungi1994.

Cohen-Kupiec R, Chet I. The molecular biology of chitin digestion. Curr Opin Biotechnol 1998;9(3):270-7.

Klein W, Gunnewiek PJA, Kowalchuk GA, Van-Veen JA. Boer, De-W., Growth of chitinolytic dune soil beta-subclass Proteobacteria in response to invading fungal hyphae. Appl Environ Microbiol 2001;67:3358-62.

Jankiewicz U, Brzezinska MS, Saks E. Identification and characterization of a chitinase of Stenotrophomonas maltophilia, a bacterium that is antagonistic towards fungal phytopathogens. Journal of bioscience and bioengineering 2012;113(1):30-5.

Weisburg WG, Barns SM, Pelletier DA, Lane DJ. 16S ribosomal DNA amplification for phylogenetic study. J Bacteriol 1991;173(2):697-703.

Lingappa Y, Lockwood JL. Chitin media for selectived culture of actinomycetes. Phytopathology 1962;52:317-23.

Bradford MM. A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 1976;72:248-54.

Laemmli UK. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 1970;227(5259):680-5.

Trudel J, Asselin A. Detection of chitinase activity after polyacrylamide gel electrophoresis. Anal Biochem 1989;178(2):362-6.

Kuk JH, Jung WJ, Jo GH, Kim YC, Kim KY, Park RD. Production of N-acetyl-beta-D-glucosamine from chitin by Aeromonas sp. GJ-18 crude enzyme. Appl Microbiol Biotechnol 2005;68(3):384-9.

Neiendam NM, Sorensen J, Fems. Chitinolytic activity of Pseudomonas fluorescens isolates from barley and sugarbeet rhizosphere. Ecol 1999;30:217-27.

Folders J, Algra J, Roelofs MS, van Loon LC, Tommassen J, Bitter W. Characterization of Pseudomonas aeruginosa chitinase, a gradually secreted protein. J Bacteriol 2001;183(24):7044-52.

Fogliano V, Ballio A, Gallo M, Woo S, Scala F, Lorito M. Pseudomonas lipodepsipeptides and fungal cell wall-degrading enzymes act synergistically in biological control. Molecular plant-microbe interactions:MPMI 2002;15(4):323-33.

Mathivanan N, Kabilan V, Murugesan K. Purification, characterization, and antifungal activity of chitinase from Fusarium chlamydosporum, a mycoparasite to groundnut rust, Puccinia arachidis. Can J Microbiol 1998;44(7):646-51.

Giambattista RD, Federici F, Petruccioli M, Fence M. The chitinolytic activity of Penicillium janthinellum P9, purification, partial characterization and potential application. J Appl Microbiol 2001;91:498-505.



How to Cite

Velusamy, P., and J. Das. “IDENTIFICATION AND CHARACTERIZATION OF ANTIFUNGAL CHITINASE FROM BACILLUS SUBTILIS JD-09 AND THEIR ROLE IN INHIBITION OF VIABLE FUNGAL GROWTH”. International Journal of Pharmacy and Pharmaceutical Sciences, vol. 6, no. 7, July 2014, pp. 232-5,



Original Article(s)