Nippon Suisan Gakkaishi 66 (2), 275-281 (2000)

Purification and Characterization of Lysozyme from Brackishwater Clam Corbicula japonica

Kouji Miyauchi,* Masahiro Matsumiya,* and Atsushi Mochizuki*

Lysozyme was purified from brackishwater clam Corbicula japonica by sequential procedures with ammonium sulfate fractionation, and CHITOPEARL BASIC BL-01 affinity, Hydroxyapatite, S-Sepharose Fast Flow, and CM-TOYOPEARL 650S column chromatographies.

The purified enzyme showed a single protein band on polyacrylamide gel electrophoresis (PAGE), and its molecular weight was estimated to be 12,000 by SDS-PAGE. Optimum pH of the enzyme was 4.8 toward Micrococcus lysodeikticus cells. The optimum temperature was 70°C. The enzyme was stable in the range of pH 3.0-6.8 and 20-90°C, respectively. The specific activity of the enzyme toward M. lysodeikticus cells was 256-fold higher than that of hen egg white lysozyme.

The N-terminal amino acid sequence and amino acid composition of the enzyme were not similar to those of hen egg white lysozyme, suggesting a genetical difference of both enzymes.


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