Cloning and sequencing of a gene encoding the 69-kDa extracellular chitinase of Janthinobacterium lividum

  1. Andrew P. Gleave1,*,
  2. Robert K. Taylor1,
  3. Bret A.M. Morris1,
  4. David R. Greenwood2

Article first published online: 17 JAN 2006

DOI: 10.1111/j.1574-6968.1995.tb07788.x

Issue

FEMS Microbiology Letters

FEMS Microbiology Letters

Volume 131, Issue 3, pages 279–288, September 1995

Additional Information

How to Cite

Gleave, A. P., Taylor, R. K., Morris, B. A. and Greenwood, D. R. (1995), Cloning and sequencing of a gene encoding the 69-kDa extracellular chitinase of Janthinobacterium lividum. FEMS Microbiology Letters, 131: 279–288. doi: 10.1111/j.1574-6968.1995.tb07788.x

Author Information

  1. 1

    Gene Transfer and Expression Group, The Horticulture and Food Research Institute of New Zealand Ltd., Private Bag 92169, Auckland, New Zealand

  2. 2

    Molecular Genetics Group, Plant Improvement Division, The Horticulture and Food Research Institute of New Zealand Ltd., Private Bag 92169, Auckland, New Zealand

* *Corresponding author. Tel.: +64 (9) 849 3660; Fax: +64 (9) 846 3330; E-mail: GleaveA@MARC.CRI.NZ

Publication History

  1. Issue published online: 17 JAN 2006
  2. Article first published online: 17 JAN 2006
  3. (Received 15 March 1995, Revised 13 June 1995, Accepted 10 July 1995)

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Keywords:

  • Janthinobacterium lividum;
  • Chitinase gene;
  • Cloning;
  • Nucleotide sequence

AbstractJanthinobacterium lividum secretes a major 56-kDa chitinase and a minor 69-kDa chitinase. A chitinase gene was defined on a 3-kb fragment of clone pRKT10, by virtue of fluorescent colonies in the presence of 4-methylumbelliferyl-β-d-N,N′,N″-chitotrioside. Nucleotide sequencing revealed an 1998-bp open reading frame with the potential to encode a 69 716-Da protein with amino acid sequences similar to those in other chitinases, suggesting it encodes the minor chitinase (Chi69). Chitinase activity of Escherichia coli (pRKTIO) lysates was detected mainly in the periplasmic fraction and immunoblotting detected a 70-kDa protein in this fraction. Chi69 has an N-terminal secretory leader peptide preceding two probable chitin-binding domains and a catalytic domain. These functional domains are separated by linker regions of proline-threonine repeats. Amino acid sequencing of cyanogen bromide cleavage-derived peptides from the major 56-kDa chitinase suggested that Chi69 may be a precursor of Chi56. In addition, an N-terminally truncated version of Chi69 retained chitinase activity as expected if in vivo processing of Chi69 generates Chi56.

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