Fig. 5.

SP-dependent extracellular alginate decomposition. a Quantification of decomposed alginate in concentrated supernatant samples (with 100 mg sodium alginate added to 5-ml concentrated supernatant protein samples; protein concentration: 0.24 g/l). GPC analysis was carried out after shaking at 120 rpm and 37 °C for 20 h. 1, JM109/pM01 (vector control); 2, JM109/pM03 (expressing wild-type AlyB of V. algivorus); 3, JM109/pM04 (SP deletion mutant); 4, JM109/pM06 (PL7 deletion mutant); 5, JM109/pM05 (CBM32 deletion mutant); 6, JM109/pM07 (SP and CBM32 deletion mutant); 7, JM109/pM02 (expressing wild-type AlyB of V. splendidus). b Average molecular weights after processing concentrated supernatant samples. Samples 1–7 are as described for panel a. c Western blot analysis of alginate in concentrated supernatants. 1, XL-Western Marker SP-2170 (Aproscience, Tokushima, Japan); 2, JM109/pM01 (vector control); 3, JM109/pM03; 4, JM109/pM04 (SP deletion mutant). d Sodium dodecyl sulphate polyacrylamide gel electrophoresis analysis alginate in concentrated supernatants. Lane 1, Novex Sharp Unstained Protein Standard (Thermo Fisher Scientific, Waltham, MA, USA); lanes 2–4 are as described for panel c