Gene cloning and characterization of a novel recombinant antifungal chitinase from papaya (carica papaya)

Yu Ting Chen, Lien Hua Hsu, I. Ping Huang, Tsun Chuang Tsai, Guan Chiun Lee, Jei Fu Shaw

Research output: Contribution to journalArticle

24 Citations (Scopus)

Abstract

A chitinase cDNA clone (CpCHI, 1002 bp) was isolated from papaya fruit, which encoded a 275 amino acid protein containing a 28 amino acid signal peptide in the N-terminal end. The predicted molecular mass of the mature protein was 26.2 kDa, and its p/value was 6.32. On the basis of its amino acid sequence homology with other plant chitinases, it was classified as a class IV chitinase. An active recombinant CpCHI enzyme was overexpressed in Escherichia coli. The purified recombinant papaya chitinase showed an optimal reaction temperature at 30 °C and a broad optimal pH ranging from 5.0 to 9.0. The recombinant enzyme was quite stable, retaining >64% activity for 3 weeks at 30 °C. The spore germination of Altemaria brassicicola could be completely inhibited by a 76 nM level of recombinant CpCHI. Recombinant CpCHI also showed antibacterial activity in which 50% of £. coli was inhibited by a 2.5 μM concentration of the enzyme.

Original languageEnglish
Pages (from-to)714-722
Number of pages9
JournalJournal of Agricultural and Food Chemistry
Volume55
Issue number3
DOIs
Publication statusPublished - 2007 Feb 7

Fingerprint

Carica
Chitinases
Cloning
papayas
chitinase
Organism Cloning
molecular cloning
Genes
Amino Acids
Enzymes
enzymes
Amino Acid Sequence Homology
amino acids
spore germination
Molecular mass
Germination
Protein Sorting Signals
Fruits
signal peptide
Spores

Keywords

  • Antifungal activity
  • Chitinase
  • Papaya
  • Recombinant expression

ASJC Scopus subject areas

  • Chemistry(all)
  • Agricultural and Biological Sciences(all)

Cite this

Gene cloning and characterization of a novel recombinant antifungal chitinase from papaya (carica papaya). / Chen, Yu Ting; Hsu, Lien Hua; Huang, I. Ping; Tsai, Tsun Chuang; Lee, Guan Chiun; Shaw, Jei Fu.

In: Journal of Agricultural and Food Chemistry, Vol. 55, No. 3, 07.02.2007, p. 714-722.

Research output: Contribution to journalArticle

Chen, Yu Ting ; Hsu, Lien Hua ; Huang, I. Ping ; Tsai, Tsun Chuang ; Lee, Guan Chiun ; Shaw, Jei Fu. / Gene cloning and characterization of a novel recombinant antifungal chitinase from papaya (carica papaya). In: Journal of Agricultural and Food Chemistry. 2007 ; Vol. 55, No. 3. pp. 714-722.
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