Biochemical properties of BbKst-kinase. Ai. Specific activity of BbKst on different substrates. The significant differences between specific activities with different substrates were determined by One-way ANOVA, ** and *** represents p < 0.001 and 0.0001, respectively. Each experiment was performed in triplicates, and the average data presented. Aii. Autoradiogram image of the substrate phosphorylation. B. pH dependent curve. Enzyme assay was performed at pH 3 to 11. The graph was fitted following Lorentzian equation. Error bars indicate standard deviation between three replicates (Bi). Respective autoradiogram of the substrate phosphorylation at different pH (Bii). C. Specific activity of BbKst-Kinase at different temperature from 4 to 45ºC using casein as substrate. Error bars indicate standard deviation between three replicates (Ci). Respective autoradiogram of the substrate phosphorylation at different pH (Cii). Di. Casein phosphorylation activity of BbKst-kinase in presence of different concentrations of Mg2+ or Mn2+ was determined in presence of 0.5, 1, 2.5, 5, 7.5 and 10 mM of Mn 2+ (source MnCl2) or Mg2+ (source MgCl2). Respective autoradiograms of phosphorylation in presemce of Mn 2+(Dii) and Mg2+(Diii).