Skip to main content
Log in

Effect of Different Carbon Sources on Endochitinase Production by Colletotrichum gloeosporioides

  • Published:
Current Microbiology Aims and scope Submit manuscript

Abstract

The present work analyzes the production of endochitinase by Colletotrichum gloeosporioides, a phytopathogenic fungus, using six different carbon sources and two pH values. For quantitative assay of endochitinase activity in solution, the synthetic substrate 4-methylumbelliferyl-β-D-N,N’,N”-triacetylchitotrioside was used. The major productions were obtained at pH 7.0 and 9.0, when colloidal chitin and glucose were used, whereas xylose and lactose were not good carbon sources. When testing different concentrations of colloidal chitin, glucose and glucosamine, colloidal chitin 0.5% was the best substrate, giving values of 2.4 U at the fifth day. When using glucose, best production occurred at 0.3% concentration, after 5 days growth, with values of 1.31 U. Endochitinase production was markedly decreased in high levels of glucose and in all glucosamine concentrations tested. SDS-PAGE co-polymerized with glycol-chitin analysis showed three major activity bands of 200, 100, and 95 kDa, when incubated at 50°C.

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

Access this article

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

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

Literature Cited

  1. E Bokma T Barends ACTV Schelting BW Dijkstra JJ Beintema (2000) ArticleTitleEnzyme kinects of hevamine, a chitinase from the rubber tree Hevea brasiliensis FEBS Lett 478 119–122 Occurrence Handle10.1016/S0014-5793(00)01833-0 Occurrence Handle10922481

    Article  PubMed  Google Scholar 

  2. Booth C (1971) Fungal culture media. In: Methods in Microbiology, Vol. 4. New York: Academic Press. pp 49–94

    Google Scholar 

  3. RC Bowers (1986) ArticleTitleCommercialization of CollegoTM: An industrialist’s view Weed Sci 34 IssueIDSuppl 24–25

    Google Scholar 

  4. M Bradford (1976) ArticleTitleA rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding Anal Biochem 72 248–254 Occurrence Handle942051

    PubMed  Google Scholar 

  5. R Cohen-Kupiec I Chet (1998) ArticleTitleThe molecular biology of chitin Curr Opin Biotechnol 9 270–277 Occurrence Handle10.1016/S0958-1669(98)80058-X Occurrence Handle9650272

    Article  PubMed  Google Scholar 

  6. J La Cruz ParticleDe A Hidalgo-Gallego JM Lora T Benitez JA Pintor-Toro (1992) ArticleTitleIsolation and characterization of three chitinases from Trichoderma harzianum Eur J Biochem 206 859–867 Occurrence Handle10.1111/j.1432-1033.1992.tb16994.x Occurrence Handle1606968

    Article  PubMed  Google Scholar 

  7. EE Deane JM Whipps JM Lynch JF Peberdy (1998) ArticleTitleThe purification and characterization of a Trichoderma harzianum exochitinase Bioch Biophy Acta 1383 101–110

    Google Scholar 

  8. GM Escott VM Hearn DJ Adams (1998) ArticleTitleInducible chitinolytic system of Aspergillus fumigatus Microbiology 144 1575–1581 Occurrence Handle9639928

    PubMed  Google Scholar 

  9. RC Gomes LTAS Sêmedo RMA Soares CS Alviano LF Linhares RRR Coelho (2000) ArticleTitleChitinolytic activity of actinomycetes from cerrado soil and their potential in biocontrol Lett Appl Microbiol 30 146–150 Occurrence Handle10.1046/j.1472-765x.2000.00687.x Occurrence Handle10736018

    Article  PubMed  Google Scholar 

  10. RC Gomes LTAS Sêmedo RMA Soares LF Linhares CJ Ulhôa CS Alviano RRR Coelho (2001) ArticleTitlePurification of a thermostable endochitinase from Streptomyces RC1071 isolated from a cerrado soil and its antagonism against phytopathogenic fungi J Appl Microbiol 90 653–661 Occurrence Handle10.1046/j.1365-2672.2001.01294.x Occurrence Handle11309080

    Article  PubMed  Google Scholar 

  11. GW Gooday W Zhu RW O’Donnel (1992) ArticleTitleWhat are the roles of chitinase in growing fungus FEMS Microbiol Lett 100 387–392 Occurrence Handle10.1016/0378-1097(92)90236-H

    Article  Google Scholar 

  12. R Gupta RK Saxena P Chaturvedi JS Virdi (1995) ArticleTitleChitinase production by Streptomyces viridificans: its potential in fungal cell wall lysis J Appl Bacteriol 78 378–383 Occurrence Handle7744723

    PubMed  Google Scholar 

  13. GE Harman CK Hayes M Lorito RM Broadway A Di Pietro C Peterbauer A Tronsmo (1993) ArticleTitleChitinolytic enzymes of Trichoderma harzianum: purification of chitobiosidase and endochitinase Mol Plant Pathol 83 313–318

    Google Scholar 

  14. Hearn VM, Escott MG, Evans EGV, David JA (1997) Intracellular and wall-associated chitinases of Aspergillus fumigatus. In: Suzuki S, Suzuki M (eds). Fungal cells in biodefense mechanism. Saikon Publishing Co., Tokyo, pp 247–252

  15. B Holmstrom-Ruddick K Mortensen (1995) ArticleTitleFactors affecting pathogenicity of a benomyl-resistant strain of Colletotrichum gloeosporioides f sp. malvae Mycol Res 99 1108–1112

    Google Scholar 

  16. M Huber E Cabib LH Miller (1991) ArticleTitleMalaria parasite chitinase and penetration of the mosquito peritrophic membrane Proc Natl Acad Sci 88 2807–2810 Occurrence Handle2011589

    PubMed  Google Scholar 

  17. C Hwang PE Kolattukudy (1995) ArticleTitleIsolation and characterization of genes expressed uniquely during appressorium formation by Colletotrichm gloeosporioides conidia induced by the host surface wax Mol Gen Genet 247 282–294 Occurrence Handle10.1007/BF00293196 Occurrence Handle7770033

    Article  PubMed  Google Scholar 

  18. C Hwang PE Kolattukudy MA Flaishman (1995) ArticleTitleCloning of a gene during appressorium formation by Colletotrichum gloeosporioides and a marked decrease in virulance by disruption of this gene Plant Cell 7 183–193 Occurrence Handle10.1105/tpc.7.2.183 Occurrence Handle7756829

    Article  PubMed  Google Scholar 

  19. J Inbar I Chet (1991) ArticleTitleEvidence that chitinases produced by Aeromonas cavie is involved in the biological control of soil-borne plant pathogens by this bacterium Soil Biol Biochem 23 973–978 Occurrence Handle10.1016/0038-0717(91)90178-M

    Article  Google Scholar 

  20. WT King LV Madden MA Ellis LL Wilson (1997) ArticleTitleEffects of temperature on sporulation and latent period of Colletotrichum spp infecting strawberry fruit. Plant Disease 81 77–84

    Google Scholar 

  21. M Lorito GE Harman CK Hayes RM Broadway A Tronsmo SL Woo A Di Pietro (1993) ArticleTitleChitinolytic enzymes produced by Trichoderma harzianum: antifungal activity of purified endochitinase and chitobiosidase Mol Plant Pathol 83 302–307

    Google Scholar 

  22. Y Luo DO TeBeest (1997) ArticleTitleInfection components of wild-type and mutant strains of Colletotrichum gloeosporioides f sp. aeschynomene on northern jointvetch Plant Disease 81 404–409

    Google Scholar 

  23. KJ McCreath GW Gooday (1992) ArticleTitleA rapid and sensitive microassay for determination of chitinolytic activity J Microbiol Methods 14 229–237 Occurrence Handle10.1016/0167-7012(92)90055-9

    Article  Google Scholar 

  24. J Monreal ET Reese (1969) ArticleTitleThe chitinase of Serratia marcescens Can J Microbiol 15 689–696 Occurrence Handle4894282

    PubMed  Google Scholar 

  25. M O’Brien RR Colwell (1987) ArticleTitleA rapid test for chitinase activity that uses 4-methylumbelliferyl-N-acetyl-β-D-glucosaminide Appl Environ Microbiol 53 1718–720 Occurrence Handle3662513

    PubMed  Google Scholar 

  26. RS Patil V Ghormade MV Deshpande (2000) ArticleTitleChitinolytic enzymes: an exploration Enz Microbiol Technol 26 473–83 Occurrence Handle10.1016/S0141-0229(00)00134-4

    Article  Google Scholar 

  27. SE Perfect HB Hughes RJ O’Connell JR Green (1999) ArticleTitleColletotrichum: A model genus for studies on pathology and fungal-plant interactions Fung Gen Biol 27 186–198 Occurrence Handle10.1006/fgbi.1999.1143

    Article  Google Scholar 

  28. HH Radwan HJ Plahner U Menger H Diekmann (1994) ArticleTitleThe 92-kDa chitinase from Streptomyces olivaceoviridis contains a lysine-C endoproteinase at its N-terminis FEMS Microbiol Lett 120 31–36 Occurrence Handle10.1016/0378-1097(94)00171-5 Occurrence Handle8056294

    Article  PubMed  Google Scholar 

  29. Y Ren KE Wee FN Chang (2000) ArticleTitleDeficiency of current methods in assaying endochitinase activity Biochem Biophys Res Commun 268 302–305 Occurrence Handle10.1006/bbrc.2000.2118 Occurrence Handle10679198

    Article  PubMed  Google Scholar 

  30. WK Roberts CP Selitreknnikoff (1988) ArticleTitlePlant and bacterial chitinases differ in antifungal activity J Gen Microbiol 134 169–176

    Google Scholar 

  31. EB Shirling D Gottlieb (1966) ArticleTitleMethods for characterization of Streptomyces species Int J Syst Bacteriol 16 312–340

    Google Scholar 

  32. RF Souza RC Gomes RRR Coelho CS Alviano RMA Soares (2003) ArticleTitlePurification and characterization of an endochitinase produced by Colletotrichum gloeosporioides FEMS Microbiol Lett 222 45–50 Occurrence Handle10.1016/S0378-1097(03)00220-9 Occurrence Handle12757945

    Article  PubMed  Google Scholar 

  33. VE Tikhonov LV Lopez-Llorca J Salinas H Jansson (2002) ArticleTitlePurification and characterization of chitinases from nematophagous fungi Verticillium chlamydosporium and V suchlasporium Fung Gen Biol 35 67–78 Occurrence Handle10.1006/fgbi.2001.1312

    Article  Google Scholar 

  34. VE Tikhonov LV Lopez-Llorca J Salinas E Monfort (2002) ArticleTitleEndochitinase activity determination using N-fluorescein-labeled chitin J Biochem Biophys Methods 60 29–38 Occurrence Handle10.1016/j.jbbm.2004.04.013

    Article  Google Scholar 

  35. A Tronsmo GE Harman (1993) ArticleTitleDetection and quantification of N-acetil-β-D-glucosaminidase, chitobiosidae, and endochitinase in solution and on gels Anal Biochem 208 74–79 Occurrence Handle10.1006/abio.1993.1010 Occurrence Handle8434798

    Article  PubMed  Google Scholar 

  36. J Trudel A Asselin (1989) ArticleTitleDetection of chitinase activity after polyacrilamide gel electrophoresis Anal Biochem 178 362–366 Occurrence Handle10.1016/0003-2697(89)90653-2 Occurrence Handle2473667

    Article  PubMed  Google Scholar 

  37. I Tsigos V Bouriotis (1995) ArticleTitlePurification and characterization of chitin deacetylase from Colletotrichum lindemuthianum J Biol Chem 270 26286–26291 Occurrence Handle10.1074/jbc.270.44.26286 Occurrence Handle7592838

    Article  PubMed  Google Scholar 

  38. H Tsujibo N Hatano T Okamoto H Endo K Miyamoto Y Inamori (1999) ArticleTitleSynthesis of chitinase in Streptomyces thermoviolaceus is regulated by a two-component sensor-regulator system FEMS Microbiol Lett 181 83–90 Occurrence Handle10.1016/S0378-1097(99)00517-0 Occurrence Handle10564792

    Article  PubMed  Google Scholar 

  39. CJ Ulhoa JF Peberdy (1991) ArticleTitlePurification and characterization of an extracellular chitobiase from Trichoderma harzianum Curr Microbiol 23 285–289

    Google Scholar 

  40. CJ Ulhoa JF Peberdy (1991) ArticleTitleRegulation of chitinase synthesis in Trichoderma harzianum J Gen Microbiol 137 2163–2169 Occurrence Handle1748872

    PubMed  Google Scholar 

  41. CJ Ulhoa JF Peberdy (1993) ArticleTitleEffect of carbon sources on chitobiase production by Trichoderma harzianum Mycol Res 97 45–4

    Google Scholar 

  42. RJ Vaidya IM Shah PR Vyas HS Chhatpar (2001) ArticleTitleProduction of chitinase and its optimization from a novel isolate Alcaligenes xylosoxydans: potential in antifungal biocontrol World J Microbiol Biotechnol 17 691–696 Occurrence Handle10.1023/A:1012927116756

    Article  Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to R.F. Souza.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Souza, R., Soares, R., Nascimento, R. et al. Effect of Different Carbon Sources on Endochitinase Production by Colletotrichum gloeosporioides. Curr Microbiol 51, 16–21 (2005). https://doi.org/10.1007/s00284-005-4506-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00284-005-4506-9

Keywords

Navigation