Vibrio pacini synthesizes multiple chitinases, of which three have been purified in this study by ammonium sulphate fractionation, chitin affinity chromatography and gel chromatography. Molecular weights of the three chitinases, Chi1, Chi2 and Chi3 are 27×103, 39×103 and 46×103 respectively, as determined by sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE). The purified enzymes have optimal activity at pH 7-8, and retain 50% enzymatic activity pH 4-9. The activities of chitinases are inhibited by Pb2+, Fe3+ and Cu2+, and increased by Ca2+, Mg2+ and Mn2+. Chi3 is found to inhibit the growth of six species of fungi. Such characters of chitinase are different from those of any other chitinase that were reported before.