Wipa SugintaArchara VongsuwanChomphunuch SongsiriritthigulHeino PrinzPeter EstibeiroRory R. DuncanJisnuson SvastiLinda A. Fothergill-GilmoreSuranaree University of TechnologyMax Planck Institut fur molekulare PhysiologieExpressOn BioSystems Ltd.University of EdinburghMahidol University2018-07-242018-07-242004-04-15Archives of Biochemistry and Biophysics. Vol.424, No.2 (2004), 171-180000398612-s2.0-1642504702https://repository.li.mahidol.ac.th/handle/20.500.14594/21198We 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.Mahidol UniversityBiochemistry, Genetics and Molecular BiologyArticleSCOPUS10.1016/j.abb.2004.01.017