Purification and characterization of extracellular beta-glucosidase from Sinorhizobium kostiense AFK-13 and its algal lytic effect on Anabaena flos-aquae.

A beta-glucosidase from the algal lytic bacterium Sinorhizobium kostiense AFK-13, grown in complex media containing cellobiose, was purified to homogeneity by successive ammonium sulfate precipitation, and anion-exchange and gel-filtration chromatographies. The enzyme was shown to be a monomeric protein with an apparent molecular mass of 52 kDa and isoelectric point of approximately 5.4. It was optimally active at pH 6.0 and 40'C and possessed a specific activity of 260.4 U/mg of protein against 4-nitrophenyl-beta-D-glucopyranoside (pNPG). A temperature-stability analysis demonstrated that the enzyme was unstable at 50 degrees C and above. The enzyme did not require divalent cations for activity, and its activity was significantly suppressed by Hg+2 and Ag+, whereas sodium dodecyl sulfate (SDS) and Triton X-100 moderately inhibited the enzyme to under 70% of its initial activity. In an algal lytic activity analysis, the growth of cyanobacteria, such as Anabaena flos-aquae, A. cylindrica, A. macrospora, Oscillatoria sancta, and Microcystis aeruginosa, was strongly inhibited by a treatment of 20 ppm/disc or 30 ppm/disc concentration of the enzyme.