Purification and Enzymatic Characteristics of Alginate Lyase from V. parahaemolyticus C20
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Abstract:
Alginate lyase was successfully purified from fermented broth of V. parahaemolyticus C20 by ultra-filtration, ammonium sulfate precipitation, ion exchange chromatography and size exclusion chromatography. The purified alginate lyase with molecular mass of 40 kD had optimal temperature of 30 ℃ and maintained 90% of its activity after 3.5-hour incubation at 30 ℃. The enzyme with optimal pH of 7.2 was stable within the pH value range of 7.0 to 7.6. Mg2+, Na+, NH4+, Ca2+, methanol and Tween-80 with certain concentration had activation of the enzyme while ethanol, acetone, DTT, urea, SDS, Zn2+, Cu2+, Fe2+ and Al3+ showed inhibition. This enzyme specifically degraded polyguluronate with Km of 1.38 mg/mL and Vmax of 0.052 mg/(ml•min).
