Fig 6.
Binding and induction of ChiC using mucin extract from porcine stomach. (A)ELISA analysis of binding between immobilized porcine stomach mucins type III and His-tagged ChiC (10 µM) and His-tagged ChiCE143Q (10 µM) detected with anti-His-tag antibody. ****P < 0.0001; ChiCE143Q versus ChiC by two-tailed Student’s t-test. Buffer was used as a control and subtracted from the samples. Data plotted represented as the mean ± SD of three replicates. (B and C) Quantification of chitinase activity of PA (PAO1) at different growth phases in LB medium and LB medium supplemented with 1% PSM type III. Activity is represented by the concentration of generated (GlcNAc)2 (µM) by the bacteria and the cell-free supernatants upon incubation with (GlcNAc)4 for 2 h after the given growth phase was reached. Purified ChiC (1 µM) was used as a positive control. The data are plotted as the mean ± SD, representing three experiments performed in biological triplicates.