Purification and Characterization of cloacae M - 1 ExtracellularAlginate Lyase from Enterobacter
نویسنده
چکیده
An alginate lyase from the cuiture supernatant of Ehterobacter cloacae M-1 rvas purified by ammonium su]fate precipitation, cation-exchange chromatography (SP-Toyopearl), and gel filtration (Ultrogel AcA44). The final preparation thus obtained showed a single band on SDS-PAGE. The purified enayme had the molecular weight of 38,ooO and 32,OOO by SDS-PAGE and gel filtration, respectiyely. The pl of the enzyme was 8.9. The optimum pH and temperature for the enzyme reaction were around 7.8 and 30eC, respectively. The enzyme was unstable on heating. EDTA complete]y inhibited the enzyme activity, but the activity was completely restored by the treatment with CaC12. The enzyme was specific for poly-guluronate and produced seyeral kinds of unsaturated oligomers from the gluluronate. This suggested that the enzyme cou]d be classified as an endo poly-guluronate lyase.
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