1887

Microbiology

Volume 146, Issue 11

The 168 gene encodes a chitosanase with similar properties to a enzyme Free

Abstract

The 168 gene encodes a chitosanase. It was found that transcription of the gene was temporally regulated and was not subject to metabolic repression. Chitosanase synthesis was abolished in a mutant strain. Csn was overproduced in , partially purified and characterized. The deduced amino acid sequence, , and optimal pH and temperature of the enzyme were closer to those of a chitosanase from sp. N174 than to those of chitosanases from other strains.

  • Received:
  • Accepted:
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  • Published Online:
Keyword(s): Bacillus subtilis , catabolite repression , chitosanase , gene expression and LR-PCR, long-range PCR
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2000-11-01
2019-12-08
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References

  1. Akiyama, K., Fujita, T., Kuroshima, K., Sakane, T., Yokota, A. & Takata, R. ( 1999; ). Purification and gene cloning of a chitosanase from Bacillus ehimensis EAG1. J Biosci Bioeng 87, 383-385.[CrossRef]
    [Google Scholar]
  2. Barnes, W. M. ( 1994; ). PCR amplification of up to 35-kb DNA with high fidelity and high yield from lambda bacteriophage templates. Proc Natl Acad Sci USA 91, 2216-2220.[CrossRef]
    [Google Scholar]
  3. Barthelemy, I., Salas, M. & Mellado, R. P. ( 1986; ). In vivo transcription of bacteriophage ϕ29 DNA: transcription initiation sites. J Virol 60, 874-879.
     [Google Scholar]
  4. Bolhuis, A., Tjalsma, H., Smith, H. E., de Jong, A., Meima, R., Venema, G., Bron, S. & van Dijl, J. M. ( 1999; ). Evaluation of bottlenecks in the late stages of protein secretion in Bacillus subtilis. Appl Environ Microbiol 65, 2934-2941.
     [Google Scholar]
  5. Boucher, I., Dupuy, A., Vidal, P., Neugebauer, W. A. & Brzezinski, R. ( 1992; ). Purification and characterization of a chitosanase from Streptomyces N174. Appl Microbiol Biotechnol 38, 188-193.
     [Google Scholar]
  6. Boucher, I., Fukamizo, T., Honda, Y., Willick, G. E., Neugebauer, W. A. & Brzezinski, R. ( 1995; ). Site-directed mutagenesis of evolutionary conserved carboxylic amino acids in the chitosanase from Streptomyces sp. N174 reveals two residues essential for catalysis. J Biol Chem 270, 31077-31082.[CrossRef]
    [Google Scholar]
  7. Bradford, M. M. ( 1976; ). A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein–dye binding. Anal Biochem 72, 248-254.[CrossRef]
    [Google Scholar]
  8. Cheng, S., Higuchi, R. & Stoneking, M. ( 1994; ). Complete mitochondrial genome amplification. Nature Genet 7, 350-351.[CrossRef]
    [Google Scholar]
  9. Christie, G. E. & Calendar, R. ( 1983; ). Bacteriophage P2 late promoters. Transcription initiation sites form two late mRNAs. J Mol Biol 167, 773-790.[CrossRef]
    [Google Scholar]
  10. Davis, B. & Eveleigh, D. E. ( 1984; ). Chitosanases: occurrence, production and immobilization. In Chitin, Chitosan and Related Enzymes, pp. 161-179. Edited by J. P. Zikakis. New York: Academic Press.
  11. Dunbar, B. S. & Schwoebel, E. D. ( 1990; ). Preparation of polyclonal antibodies. Methods Enzymol 182, 663-670.
     [Google Scholar]
  12. Dygert, S., Li, L. H, Florida, D. & Thoma, J. A. ( 1965; ). Determination of reducing sugar with improved precision. Anal Biochem 13, 367-374.[CrossRef]
    [Google Scholar]
  13. Edman, M., Jarhede, T., Sjöström, M. & Wieslander, A. ( 1999; ). Different sequence patterns in signal peptides from Mycoplasmas, other Gram-positive bacteria and Escherichia coli: a multivariate data analysis. Proteins 35, 195-205.[CrossRef]
    [Google Scholar]
  14. Harwood, C. R. & Cutting, S. M. (1990). Molecular Biological Methods for Bacillus. Chichester: Wiley.
  15. Hirano, S. ( 1996; ). Chitin biotechnology applications. Biotechnol Annu Rev 2, 237-258.
     [Google Scholar]
  16. Kedziersdi, W. & Porter, J. C. ( 1991; ). A novel non-enzymatic procedure for removing DNA template from RNA transcription mixtures. Biotechniques 10, 210-214.
     [Google Scholar]
  17. Kyte, J. & Doolittle, R. F. ( 1982; ). A simple method for displaying the hydropathic character of a protein. J Mol Biol 157, 105-132.[CrossRef]
    [Google Scholar]
  18. Laemmli, U. K. ( 1970; ). Cleavage of structural proteins during the assembly of the head of bacteriophage T4. Nature 227, 680-685.[CrossRef]
    [Google Scholar]
  19. Leloup, L., Haddaoui, E. A., Chambert, R. & Petit-Glatron, M. F. ( 1997; ). Characterization of the rate-limiting step of the secretion of Bacillus subtilis α-amylase overproduced during the exponential phase of growth. Microbiology 143, 3295-3303.[CrossRef]
    [Google Scholar]
  20. Lu, Z., Li, Y., Que, Q., Kutish, G. F., Rock, D. L. & Von Etten, J. L. ( 1996; ). Analysis of 94 kb of the chlorella virus PBCV-1 330-kb genome: map positions 88 to 182. Virology 216, 102-123.[CrossRef]
    [Google Scholar]
  21. Marcotte, E. M., Monzingo, A. F., Ernst, S. R., Brzezinski, R. & Robertus, J. D. ( 1996; ). X-ray structure of an anti-fungal chitosanse from Streptomyces N174. Nature Struct Biol 3, 155-162.[CrossRef]
    [Google Scholar]
  22. Masson, J. Y., Boucher, I., Neugebauer, W. A., Ramotar, D. & Brzezinski, R. ( 1995; ). A new chitosanase gene from a Nocardioides sp. is a third member of glycosyl hydrolase family 46. Microbiology 141, 2629-2635.[CrossRef]
    [Google Scholar]
  23. Maxam, A. M. & Gilbert, W. ( 1980; ). Sequencing end-labeled DNAs with base-specific chemical cleavages. Methods Enzymol 65, 499-560.
     [Google Scholar]
  24. Mellado, R. P., Delius, H., Klein, B. & Murray, K. ( 1981; ). Transcription of sea urchin histone genes in Escherichia coli. Nucleic Acids Res 9, 3889-3906.[CrossRef]
    [Google Scholar]
  25. Mellado, R. P, Barthelemy, I. & Salas, M. ( 1986; ). In vivo transcription of bacteriophage ϕ29 DNA early and late promoter sequences. J Mol Biol 191, 191-197.[CrossRef]
    [Google Scholar]
  26. Miwa, Y., Nakata, A., Ogiwara, A., Yamamoto, M. & Fujita, Y. ( 2000; ). Evaluation and characterization of catabolite-responsive elements (cre) of Bacillus subtilis. Nucleic Acids Res 28, 1206-1210.[CrossRef]
    [Google Scholar]
  27. Park, J. K., Shimono, K., Ochiai, N., Shigeru, K., Kurita, M., Ohta, Y., Tanaka, K., Matsuda, H. & Kawamukai, M. ( 1999; ). Purification, characterization, and gene analysis of a chitosanase (ChoA) from Matsuebacter chitosanotabidus 3001. J Bacteriol 181, 6642-6649.
     [Google Scholar]
  28. Parro, V. & Mellado, R. P. ( 1993; ). Heterologous recognition in vivo of promoter sequences from the Streptomyces coelicolor dagA gene. FEMS Microbiol Lett 106, 347-356.[CrossRef]
    [Google Scholar]
  29. Parro, V. & Mellado, R. P. ( 1994; ). Effect of glucose on agarase overproduction by Streptomyces. Gene 145, 49-55.[CrossRef]
    [Google Scholar]
  30. Parro, V., San Román, M., Galindo, I., Purnelle, B., Bolotin, A., Sorokin, A. & Mellado, R. P. ( 1997a; ). A 23911 bp region of the Bacillus subtilis genome comprising genes located upstream and downstream of the lev operon. Microbiology 143, 1321-1326.[CrossRef]
    [Google Scholar]
  31. Parro, V., Vives, C., Godia, F. & Mellado, R. P. ( 1997b; ). Overproduction and purification of an agarase of bacterial origin. J Biotechnol 58, 59-66.[CrossRef]
    [Google Scholar]
  32. Parro, V., Mellado, R. P. & Harwood, C. R. ( 1998; ). Effect of phosphate limitation on agarase production by Streptomyces lividans TK21. FEMS Microbiol Lett 158, 107-113.
     [Google Scholar]
  33. Pelletier, A. & Sygush, J. ( 1990; ). Purification and characterization of three chitosanase activities from Bacillus megaterium P1. Appl Environ Microbiol 56, 844-848.
     [Google Scholar]
  34. Price, J. S. & Storck, S. ( 1975; ). Production, purification and characterization of an extracellular chitosanase from Streptomyces. J Bacteriol 124, 1574-1585.
     [Google Scholar]
  35. Saito, J. I., Kita, A., Higuchi, Y., Nagat, Y., Ando, A. & Miki, K. ( 1999; ). Crystal stucture of chitosanase from Bacillus circulans MH-K1 at 1·6-Å resolution and its substrate recognition mechanism. J Biol Chem 43, 30818-30825.
     [Google Scholar]
  36. Sambrook, J., Fritsch, E. F. & Maniatis, T. (1989). Molecular Cloning: a Laboratory Manual, 2nd edn. Cold Spring Harbor, NY: Cold Spring Harbor Laboratory.
  37. Sandorf, P. A. ( 1989; ). Chitosan: commercial uses and potential applications. In Chitin and Chitosan, pp. 51-69. Edited by G. Skjak-Braek, T. Anthonsen & P. A. Sandorf. London: Elsevier.
  38. Shimosaka, M., Kumehara, M., Zhang, X. Y., Nogawa, M. & Okazaki, M. ( 1996; ). Cloning and characterization of a chitosanase gene from the plant pathogenic fungus Fusarium solani. J Ferment Bioeng 82, 426-431.[CrossRef]
    [Google Scholar]
  39. Somashekar, D. & Joseph, R. ( 1996; ). Chitosanases – properties and applications: a review. Bioresour Technol 55, 35-45.[CrossRef]
    [Google Scholar]
  40. Swofford, D. L. (1988). Phylogenetic Analysis Using Parsimony (paup), version 4.0. Sinauer Associates, Sunderland, MA, USA.
  41. Thompson, J. D., Higgins, D. G. & Gibson, T. J. ( 1994; ). clustal w: improving the sensitivity of progressive multiple sequence alignment though sequence weighting, position-specific gap penalties and weight matrix choice. Nucleic Acids Res 22, 4673-4680.[CrossRef]
    [Google Scholar]
  42. Timmons, T. M. & Dunbar, B. S. ( 1990; ). Protein blotting and immunodetection. Methods Enzymol 182, 679-688.
     [Google Scholar]
  43. Yabuki, M., Uchiyama, A., Suzuki, K., Ando, A. & Fujii, T. ( 1988; ). Purification and properties of chitosanase from Bacillus circulans MH-K1. J Gen Appl Microbiol 34, 255-270.[CrossRef]
    [Google Scholar]
  44. Yamada, T., Hiramatsu, S., Songsri, P. & Fujie, M. ( 1997; ). Alternative expression of a chitosanase gene produces two different proteins in cells infected with Chlorella virus CVK2. Virology 230, 361-368.[CrossRef]
    [Google Scholar]
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The Bacillus subtilis 168 csn gene encodes a chitosanase with similar properties to a Streptomyces enzyme
Microbiology 146, 2929 (2000); https://doi.org/10.1099/00221287-146-11-2929
/content/journal/micro/10.1099/00221287-146-11-2929
/content/journal/micro/10.1099/00221287-146-11-2929
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