Fig. 3: Functional validation of three representative P-acquisition AMGs.

a, e, i show the computational protein models of two pyrophosphatases (i.e., PPa1 encoded by the AMG ppa1 and PPa54 encoded by the AMG ppa54) and one alkaline phosphatase (i.e., PhoD22 encoded by the AMG phoD22), respectively. Helices and sheets are colored in a rainbow scheme (from the N terminus in red to the C terminus in blue). Detailed information on individual computational protein models is provided in Supplementary Data 12. b, f, j show the activities of PPa1, PPa54, and PhoD22, respectively, with comparisons between each phage-encoded protein and its corresponsive controls. P1: the commercial pyrophosphatase (PPa, 1000 U mL⁻¹) of Escherichia coli was used as a positive control for both PPa1 and PPa54. N: the total protein from the recombinant E.coli cells transformed with an empty pET28a vector was used as a negative control for all three phage-encoded proteins. P2: the commercial recombinant E.coli alkaline phosphatase (1000 U mL⁻¹) was used as a positive control for PhoD22. The dots overlaying each bar represent the corresponding data points. c, g, k show the effects of pH on the activities of PPa1, PPa54, and PhoD22, respectively. d, h, l show the effects of temperature on the activities of PPa1, PPa54, and PhoD22, respectively. Data presented in (b–d, f–h, j–l) were mean values ± standard deviations from three independent experiments (i.e., n = 3). Relative activities of a given phage-encoded protein shown in individual panels (c, d, g, h, k, l) were calculated based on the highest activity of that protein reported within the corresponsive panel.