Nippon Suisan Gakkaishi 80 (3), 371-378 (2014)

Isolation of alginate lyase-producing actinomycetes from the marine environment and purification of lyase

CHIAKI IMADA,* KEI SAKATA, TAKESHI TERAHARA AND TAKESHI KOBAYASHI

Graduate School of Marine Science and Technology, Tokyo University of Marine Science and Technology, Minato, Tokyo 108-8477, Japan

 A total of 605 actinomycetes were isolated from marine and terrestrial sediments of various Japanese coastal areas using several isolation media. The largest number of alginate lyase (A.L.)-producing strains were isolated from marine sediment samples collected in Otsuchi Bay in Iwate Prefecture. The activities of 18 A.L.-producing strains were measured using liquid culture media and were identified by 16S rDNA sequencing analysis. Among the 18 strains, one species belonging to the genus of Micromonospora (named as strain OB40608, isolated from Otsuchi Bay at a depth of 100 m) showed high A.L. activity and has never been reported as an A.L. producer, therefore it was selected for further study. A.L. was purified from culture supernatant of strain OB40608 by ammonium sulfate precipitation, cation exchange chromatography (Cellufine C-500 m) and gel filtration chromatography (Sephacryl S-200 HR). The final preparation showed a single band on polyacryl amide gel electrophoresis. The molecular weight of purified A.L. was 114 kDa, and the optimum temperature and pH were 50℃ and pH 7.0, respectively.


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