Skip to main content
PMC home page
Journal of Bacteriology logoLink to Journal of Bacteriology
. 1996 Sep;178(17):5065–5070. doi: 10.1128/jb.178.17.5065-5070.1996

A modular family 19 chitinase found in the prokaryotic organism Streptomyces griseus HUT 6037.

T Ohno 1, S Armand 1, T Hata 1, N Nikaidou 1, B Henrissat 1, M Mitsutomi 1, T Watanabe 1
PMCID: PMC178299  PMID: 8752320

Abstract

The specificity of chitinase C-1 of Streptomyces griseus HUT 6037 for the hydrolysis of the beta-1,4-glycosidic linkages in partially acetylated chitosan is different from that of other microbial chitinases. In order to study the primary structure of this unique chitinase, the chiC gene specifying chitinase C-1 was cloned and its nucleotide sequence was determined. The gene encodes a polypeptide of 294 amino acids with a calculated size of 31.4 kDa. Comparison of the amino acid sequence of the deduced polypeptide with that of other proteins revealed a C-terminal catalytic domain displaying considerable sequence similarity to the catalytic domain of plant class I, II, and IV chitinases which form glycosyl hydrolase family 19. The N-terminal domain of the deduced polypeptide exhibits sequence similarity to substrate-binding domains of several microbial chitinases and cellulases but not to the chitin-binding domains of plant chitinases. The previously purified chitinase C-1 from S. griseus is suggested to be generated by proteolytic removal of the N-terminal chitin-binding domain and corresponds to the catalytic domain of the chitinase encoded by the chiC gene. High-performance liquid chromatography analysis of the hydrolysis products from N-acetyl chitotetraose revealed that chitinase C-1 catalyzes hydrolysis of the glycosidic bond with inversion of the anomeric configuration, in agreement with the previously reported inverting mechanism of plant class I chitinases. This is the first report of a family 19 chitinase found in an organism other than higher plants.

Full Text

The Full Text of this article is available as a PDF (400.2 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Ames G. F. Resolution of bacterial proteins by polyacrylamide gel electrophoresis on slabs. Membrane, soluble, and periplasmic fractions. J Biol Chem. 1974 Jan 25;249(2):634–644. [PubMed] [Google Scholar]
  2. Armand S., Tomita H., Heyraud A., Gey C., Watanabe T., Henrissat B. Stereochemical course of the hydrolysis reaction catalyzed by chitinases A1 and D from Bacillus circulans WL-12. FEBS Lett. 1994 Apr 25;343(2):177–180. doi: 10.1016/0014-5793(94)80314-5. [DOI] [PubMed] [Google Scholar]
  3. Blaak H., Schnellmann J., Walter S., Henrissat B., Schrempf H. Characteristics of an exochitinase from Streptomyces olivaceoviridis, its corresponding gene, putative protein domains and relationship to other chitinases. Eur J Biochem. 1993 Jun 15;214(3):659–669. doi: 10.1111/j.1432-1033.1993.tb17966.x. [DOI] [PubMed] [Google Scholar]
  4. Davies G., Henrissat B. Structures and mechanisms of glycosyl hydrolases. Structure. 1995 Sep 15;3(9):853–859. doi: 10.1016/S0969-2126(01)00220-9. [DOI] [PubMed] [Google Scholar]
  5. Fukamizo T., Honda Y., Goto S., Boucher I., Brzezinski R. Reaction mechanism of chitosanase from Streptomyces sp. N174. Biochem J. 1995 Oct 15;311(Pt 2):377–383. doi: 10.1042/bj3110377. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Gebler J., Gilkes N. R., Claeyssens M., Wilson D. B., Béguin P., Wakarchuk W. W., Kilburn D. G., Miller R. C., Jr, Warren R. A., Withers S. G. Stereoselective hydrolysis catalyzed by related beta-1,4-glucanases and beta-1,4-xylanases. J Biol Chem. 1992 Jun 25;267(18):12559–12561. [PubMed] [Google Scholar]
  7. Gilkes N. R., Henrissat B., Kilburn D. G., Miller R. C., Jr, Warren R. A. Domains in microbial beta-1, 4-glycanases: sequence conservation, function, and enzyme families. Microbiol Rev. 1991 Jun;55(2):303–315. doi: 10.1128/mr.55.2.303-315.1991. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. Gleave A. P., Taylor R. K., Morris B. A., Greenwood D. R. Cloning and sequencing of a gene encoding the 69-kDa extracellular chitinase of Janthinobacterium lividum. FEMS Microbiol Lett. 1995 Sep 15;131(3):279–288. doi: 10.1111/j.1574-6968.1995.tb07788.x. [DOI] [PubMed] [Google Scholar]
  9. Hart P. J., Pfluger H. D., Monzingo A. F., Hollis T., Robertus J. D. The refined crystal structure of an endochitinase from Hordeum vulgare L. seeds at 1.8 A resolution. J Mol Biol. 1995 Apr 28;248(2):402–413. [PubMed] [Google Scholar]
  10. Henrissat B. A classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J. 1991 Dec 1;280(Pt 2):309–316. doi: 10.1042/bj2800309. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Henrissat B., Bairoch A. New families in the classification of glycosyl hydrolases based on amino acid sequence similarities. Biochem J. 1993 Aug 1;293(Pt 3):781–788. doi: 10.1042/bj2930781. [DOI] [PMC free article] [PubMed] [Google Scholar]
  12. Henrissat B. Weak sequence homologies among chitinases detected by clustering analysis. Protein Seq Data Anal. 1990 Dec;3(6):523–526. [PubMed] [Google Scholar]
  13. Iseli B., Armand S., Boller T., Neuhaus J. M., Henrissat B. Plant chitinases use two different hydrolytic mechanisms. FEBS Lett. 1996 Mar 11;382(1-2):186–188. doi: 10.1016/0014-5793(96)00174-3. [DOI] [PubMed] [Google Scholar]
  14. Kuranda M. J., Robbins P. W. Chitinase is required for cell separation during growth of Saccharomyces cerevisiae. J Biol Chem. 1991 Oct 15;266(29):19758–19767. [PubMed] [Google Scholar]
  15. LOWRY O. H., ROSEBROUGH N. J., FARR A. L., RANDALL R. J. Protein measurement with the Folin phenol reagent. J Biol Chem. 1951 Nov;193(1):265–275. [PubMed] [Google Scholar]
  16. Laemmli U. K. Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature. 1970 Aug 15;227(5259):680–685. doi: 10.1038/227680a0. [DOI] [PubMed] [Google Scholar]
  17. Lipman D. J., Pearson W. R. Rapid and sensitive protein similarity searches. Science. 1985 Mar 22;227(4693):1435–1441. doi: 10.1126/science.2983426. [DOI] [PubMed] [Google Scholar]
  18. Matsudaira P. Sequence from picomole quantities of proteins electroblotted onto polyvinylidene difluoride membranes. J Biol Chem. 1987 Jul 25;262(21):10035–10038. [PubMed] [Google Scholar]
  19. Mitsutomi M., Kidoh H., Tomita H., Watanabe T. The action of Bacillus circulans WL-12 chitinases on partially N-acetylated chitosan. Biosci Biotechnol Biochem. 1995 Mar;59(3):529–531. doi: 10.1271/bbb.59.529. [DOI] [PubMed] [Google Scholar]
  20. Mitsutomi M., Ohtakara A., Fukamizo T., Goto S. Action pattern of Aeromonas hydrophila chitinase on partially N-acetylated chitosan. Agric Biol Chem. 1990 Apr;54(4):871–877. [PubMed] [Google Scholar]
  21. Molano J., Durán A., Cabib E. A rapid and sensitive assay for chitinase using tritiated chitin. Anal Biochem. 1977 Dec;83(2):648–656. doi: 10.1016/0003-2697(77)90069-0. [DOI] [PubMed] [Google Scholar]
  22. Molano J., Polacheck I., Duran A., Cabib E. An endochitinase from wheat germ. Activity on nascent and preformed chitin. J Biol Chem. 1979 Jun 10;254(11):4901–4907. [PubMed] [Google Scholar]
  23. Ohtakara A., Matsunaga H., Mitsutomi M. Action pattern of Streptomyces griseus chitinase on partially N-acetylated chitosan. Agric Biol Chem. 1990 Dec;54(12):3191–3199. [PubMed] [Google Scholar]
  24. Perrakis A., Tews I., Dauter Z., Oppenheim A. B., Chet I., Wilson K. S., Vorgias C. E. Crystal structure of a bacterial chitinase at 2.3 A resolution. Structure. 1994 Dec 15;2(12):1169–1180. doi: 10.1016/s0969-2126(94)00119-7. [DOI] [PubMed] [Google Scholar]
  25. Roberts R. L., Cabib E. Serratia marcescens chitinase: one-step purification and use for the determination of chitin. Anal Biochem. 1982 Dec;127(2):402–412. doi: 10.1016/0003-2697(82)90194-4. [DOI] [PubMed] [Google Scholar]
  26. Sidhu S. S., Kalmar G. B., Willis L. G., Borgford T. J. Streptomyces griseus protease C. A novel enzyme of the chymotrypsin superfamily. J Biol Chem. 1994 Aug 5;269(31):20167–20171. [PubMed] [Google Scholar]
  27. Sitrit Y., Vorgias C. E., Chet I., Oppenheim A. B. Cloning and primary structure of the chiA gene from Aeromonas caviae. J Bacteriol. 1995 Jul;177(14):4187–4189. doi: 10.1128/jb.177.14.4187-4189.1995. [DOI] [PMC free article] [PubMed] [Google Scholar]
  28. Strohl W. R. Compilation and analysis of DNA sequences associated with apparent streptomycete promoters. Nucleic Acids Res. 1992 Mar 11;20(5):961–974. doi: 10.1093/nar/20.5.961. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Takayanagi T., Ajisaka K., Takiguchi Y., Shimahara K. Isolation and characterization of thermostable chitinases from Bacillus licheniformis X-7u. Biochim Biophys Acta. 1991 Jul 12;1078(3):404–410. doi: 10.1016/0167-4838(91)90163-t. [DOI] [PubMed] [Google Scholar]
  30. Terwisscha van Scheltinga A. C., Armand S., Kalk K. H., Isogai A., Henrissat B., Dijkstra B. W. Stereochemistry of chitin hydrolysis by a plant chitinase/lysozyme and X-ray structure of a complex with allosamidin: evidence for substrate assisted catalysis. Biochemistry. 1995 Dec 5;34(48):15619–15623. doi: 10.1021/bi00048a003. [DOI] [PubMed] [Google Scholar]
  31. Terwisscha van Scheltinga A. C., Kalk K. H., Beintema J. J., Dijkstra B. W. Crystal structures of hevamine, a plant defence protein with chitinase and lysozyme activity, and its complex with an inhibitor. Structure. 1994 Dec 15;2(12):1181–1189. doi: 10.1016/s0969-2126(94)00120-0. [DOI] [PubMed] [Google Scholar]
  32. Watanabe T., Ito Y., Yamada T., Hashimoto M., Sekine S., Tanaka H. The roles of the C-terminal domain and type III domains of chitinase A1 from Bacillus circulans WL-12 in chitin degradation. J Bacteriol. 1994 Aug;176(15):4465–4472. doi: 10.1128/jb.176.15.4465-4472.1994. [DOI] [PMC free article] [PubMed] [Google Scholar]
  33. Watanabe T., Kobori K., Miyashita K., Fujii T., Sakai H., Uchida M., Tanaka H. Identification of glutamic acid 204 and aspartic acid 200 in chitinase A1 of Bacillus circulans WL-12 as essential residues for chitinase activity. J Biol Chem. 1993 Sep 5;268(25):18567–18572. [PubMed] [Google Scholar]
  34. Watanabe T., Oyanagi W., Suzuki K., Ohnishi K., Tanaka H. Structure of the gene encoding chitinase D of Bacillus circulans WL-12 and possible homology of the enzyme to other prokaryotic chitinases and class III plant chitinases. J Bacteriol. 1992 Jan;174(2):408–414. doi: 10.1128/jb.174.2.408-414.1992. [DOI] [PMC free article] [PubMed] [Google Scholar]
  35. Watanabe T., Oyanagi W., Suzuki K., Tanaka H. Chitinase system of Bacillus circulans WL-12 and importance of chitinase A1 in chitin degradation. J Bacteriol. 1990 Jul;172(7):4017–4022. doi: 10.1128/jb.172.7.4017-4022.1990. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Yamada H., Imoto T. A convenient synthesis of glycolchitin, a substrate of lysozyme. Carbohydr Res. 1981 May 18;92(1):160–162. doi: 10.1016/s0008-6215(00)85993-5. [DOI] [PubMed] [Google Scholar]

Articles from Journal of Bacteriology are provided here courtesy of American Society for Microbiology (ASM)

RESOURCES