1887

Abstract

The gene encoding a major, inducible 45 kDa chitinase of was cloned and analysis of the deduced amino acid sequence identified a chitinase of the fungal/bacterial class which was designated ChiB1. Recombinant ChiB1, expressed in , was shown to function by a retaining mechanism of action. That is, the -conformation of the chitin substrate linkage was preserved in the product in a manner typical of family 18 chitinases. Cleavage patterns with the -acetylglucosamine (GlcNAc) oligosaccharide substrates GlcNAc, GlcNAc and GlcNAc indicated that the predominant reaction involved hydrolysis of GlcNAc from the non-reducing end of each substrate. Products of transglycosylation were also identified in each incubation. Following disruption of by gene replacement, growth and morphology of disruptants and of the wild-type strain were essentially identical. However, during the autolytic phase of batch cultures the level of chitinase activity in culture filtrate from a disruptant was much lower than the activity from the wild-type. The search for chitinases with morphogenetic roles in filamentous fungi should perhaps focus on chitinases of the fungal/plant class although such an investigation will be complicated by the identification of at least 11 putative active site domains for family 18 chitinases in the TIGR database (http://www.tigr.org/).

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2003-10-01
2024-04-26
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References

  1. Andriole V. T. 2000; Current and future antifungal therapy: new targets for antifungal therapy. Int J Antimicrob Agents 16:317–321
    [Google Scholar]
  2. Blaiseau P.-L., Lafay J.-F. 1992; Primary structure of a chitinase-encoding gene ( chi1 ) from the filamentous fungus Aphanocladium album : similarity to bacterial chitinases. Gene 120:243–248
    [Google Scholar]
  3. Bruneau J.-M., Magnin T., Tagat E., Legrand R., Bernard M., Diaquin M., Fudali C., Latgé J.-P. 2001; Proteome analysis of Aspergillus fumigatus identifies glycosylphosphatidylinositol-anchored proteins associated to the cell wall biosynthesis. Electrophoresis 22:2812–2823
    [Google Scholar]
  4. Denning D. W. 1998; Invasive aspergillosis. Clin Infect Dis 26:781–805
    [Google Scholar]
  5. Dickinson K., Keer V., Hitchcock C. A., Adams D. J. 1991; Microsomal chitinase from Candida albicans . Biochim Biophys Acta 1073:177–182
    [Google Scholar]
  6. Escott G. M., Hearn V. M., Adams D. J. 1998; Inducible chitinolytic system of Aspergillus fumigatus . Microbiology 144:1575–1581
    [Google Scholar]
  7. Frieman M. B., McCaffery J. M., Cormack B. P. 2002; Modular domain structure in the Candida glabrata adhesin Epa1p, a β 1,6 glucan-cross-linked cell wall protein. Mol Microbiol 46:479–492
    [Google Scholar]
  8. Fukamizo T., Sasaki C., Schelp E., Bortone K., Robertus J. D. 2001; Kinetic properties of chitinase-1 from the fungal pathogen Coccidioides immitis . Biochemistry 40:2448–2454
    [Google Scholar]
  9. Hayes C. K., Klemsdal S., Lorito M., Di Pietro A., Peterbauer C., Nakas J. P., Tronsmo A., Harman G. E. 1994; Isolation and sequence of an endochitinase-encoding gene from a cDNA library of Trichoderma harzianum . Gene 138:143–148
    [Google Scholar]
  10. Hearn V. M., Escott G. M., Evans E. G. V., Adams D. J. 1998; Complex chitinolytic system of Aspergillus fumigatus . Microbios 93:85–104
    [Google Scholar]
  11. Henrissat B. 1999; Classification of chitinase modules. In Chitin and Chitinases pp  137–156 Edited by Jollès P., Muzzarelli R. A. A. Basel, Switzerland: Burkhäuser;
    [Google Scholar]
  12. Kim D.-J., Baek J.-M., Uribe P., Kenerley C. M., Cook D. R. 2002; Cloning and characterization of multiple glycosyl hydrolase genes from Trichoderma virens . Curr Genet 40:374–384
    [Google Scholar]
  13. Koga D., Yoshioka T., Arakane Y. 1998; HPLC analysis of anomeric formation and cleavage pattern by chitinolytic enzyme. Biosci Biotechnol Biochem 62:1643–1646
    [Google Scholar]
  14. Kuranda M. J., Robbins P. W. 1991; Chitinase is required for cell separation during growth of Saccharomyces cerevisiae . J Biol Chem 266:19758–19767
    [Google Scholar]
  15. Latgé J.-P. 2001; The pathobiology of Aspergillus fumigatus . Trends Microbiol 9:382–389
    [Google Scholar]
  16. McCreath K. J., Gooday G. W. 1992; A rapid and sensitive microassay for determination of chitinolytic activity. J Microbiol Methods 14:229–237
    [Google Scholar]
  17. Mouyna I., Fontaine T., Vai M., Monod M., Fonzi W. A., Diaquin M., Popolo L., Hartland R. P., Latgé J.-P. 2000; Glycosylphosphatidylinositol-anchored glucanosyltransferases play an active role in the biosynthesis of the fungal cell wall. J Biol Chem 275:14882–14889
    [Google Scholar]
  18. Paris S. 1994; Isolation of protease negative mutants of Aspergillus fumigatus by insertion of a disrupted gene. In Molecular Biology of Pathogenic Fungi. A Laboratory Manual pp  49–55 Edited by Maresca B., Kobayashi G. S. New York: Telos Press;
    [Google Scholar]
  19. Pishko E. J., Kirkland T. N., Cole G. T. 1995; Isolation and characterization of two chitinase-encoding genes ( cts1 , cts2 ) from the fungus Coccidioides immitis . Gene 167:173–177
    [Google Scholar]
  20. Punt P. J., Oliver R. P., Dingemanse M. A., Pouwels P. H., van den Hondel C. A. M. J. J. 1987; Transformation of Aspergillus based on the hygromycin B resistance marker from Escherichia coli . Gene 56:117–124
    [Google Scholar]
  21. Rast D. M., Horsch M., Furter R., Gooday G. W. 1991; A complex chitinolytic system in exponentially growing mycelium of Mucor rouxii : properties and function. J Gen Microbiol 137:2797–2810
    [Google Scholar]
  22. Reichard U., Hung C.-Y., Thomas P. W., Cole G. T. 2000; Disruption of the gene which encodes a serodiagnostic antigen and chitinase of the human fungal pathogen Coccidioides immitis . Infect Immun 68:5830–5838
    [Google Scholar]
  23. Robertus J. D., Monzingo A. F. 1999; The structure and action of chitinases. In Chitin and Chitinases pp  125–135 Edited by Jollès P., Muzzarelli R. A. A. Basel, Switzerland: Burkhäuser;
    [Google Scholar]
  24. Sambrook J., Fritsch E. F., Maniatis T. 1989 Molecular Cloning: a Laboratory Manual , 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory;
    [Google Scholar]
  25. Takaya N., Yamazaki D., Horiuchi H., Ohta A., Takagi M. 1998a; Cloning and characterization of a chitinase-encoding gene ( chiA ) from Aspergillus nidulans , disruption of which decreases germination frequency and hyphal growth. Biosci Biotechnol Biochem 62:60–65
    [Google Scholar]
  26. Takaya N., Yamazaki D., Horiuchi H., Ohta A., Takagi M. 1998b; Intracellular chitinase gene from Rhizopus oligosporus : molecular cloning and characterization. Microbiology 144:2647–2654
    [Google Scholar]
  27. Vogel H. J. 1964; Distribution of lysine pathways among fungi: evolutionary implications. Am Nat 98:435–446
    [Google Scholar]
  28. Xia G., Jin C., Zhou J., Yang S., Zhang S., Jin C. 2001; A novel chitinase having a unique mode of action from Aspergillus fumigatus YJ-407. Eur J Biochem 268:4079–4085
    [Google Scholar]
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