Cloning, Expression and Characterization of an Alginate Lyase in Bacillus subtilis WB600

The aim of this study was to further broaden the heterologous expression alginate lyase from Vibrio alginolyticus in a Bacillus subtilis vector. A B. WB600/pP43NMK-alg62 strain constructed. (NH4)2SO4 precipitation and Ni-affinity chromatography were performed purify enzyme. We then characterized Its molecular weight 57.64 kDa, it worked optimally at 30 °C with pH 8.0. Ca2+ markedly enhanced enzymatic activity Alg62 while Cu2+ Ni2+ inhibited its activity. had wide range substrate specificity, showing high toward sodium polyG. Following optimization fermentation process, optimal conditions for recombinant as follows: temperature 37 °C, 7.0, medium consisting glycerol 15 g/L, yeast powder 25 g/L K+ 1.5 mmol/L. At these conditions, enzyme reached 318.21 U/mL, which 1.54 times higher than initial