Publication: An endochitinase A from Vibrio carchariae: Cloning, expression, mass and sequence analyses, and chitin hydrolysis
Issued Date
2004-04-15
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ISSN
00039861
Other identifier(s)
2-s2.0-1642504702
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Mahidol University
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SCOPUS
Bibliographic Citation
Archives of Biochemistry and Biophysics. Vol.424, No.2 (2004), 171-180
Suggested Citation
Wipa Suginta, Archara Vongsuwan, Chomphunuch Songsiriritthigul, Heino Prinz, Peter Estibeiro, Rory R. Duncan, Jisnuson Svasti, Linda A. Fothergill-Gilmore An endochitinase A from Vibrio carchariae: Cloning, expression, mass and sequence analyses, and chitin hydrolysis. Archives of Biochemistry and Biophysics. Vol.424, No.2 (2004), 171-180. doi:10.1016/j.abb.2004.01.017 Retrieved from: https://repository.li.mahidol.ac.th/handle/20.500.14594/21198
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Title
An endochitinase A from Vibrio carchariae: Cloning, expression, mass and sequence analyses, and chitin hydrolysis
Abstract
We provide evidence that chitinase A from Vibrio carchariae acts as an endochitinase. The chitinase A gene isolated from V. carchariae genome encodes 850 amino acids expressing a 95-kDa precursor. Peptide masses of the native enzyme identified from MALDI-TOF or nanoESIMS were identical with the putative amino acid sequence translated from the corresponding nucleotide sequence. The enzyme has a highly conserved catalytic TIM-barrel region as previously described for Serratia marcescens ChiA. The Mrof the native chitinase A was determined to be 62,698, suggesting that the C-terminal proteolytic cleavage site was located between R597and K598. The DNA fragment that encodes the processed enzyme was subsequently cloned and expressed in Escherichia coli. The expressed protein exhibited chitinase activity on gel activity assay. Analysis of chitin hydrolysis using HPLC/ESI-MS confirmed the endo characteristics of the enzyme. © 2004 Elsevier Inc. All rights reserved.