Extended Data Fig. 5: Selection of the improved PAM mono-variants from the site-saturation mutagenesis strategy of seven PAM residues (1st level of multilevel screening).
From: An engineered enzyme embedded into PLA to make self-biodegradable plastic
Boxplot distribution of the relative halo diameter sizes of the independent site-saturation mutagenesis of the seven residues of PAM (S101, S103, T105, T106, G133, E159 and I217) as compared to wild type PAM halo. Halo diameter sizes were evaluated after incubation at 45 °C of agarose-immobilized PLA substrate, buffered at pH 9.0 loaded with protein extracts. Individual boxes were drawn using first and third quartiles of the distribution, the median value is shown as an orange line. Boundaries of the whiskers are based on a 1.5 interquartile range value and outliers are shown (open circle). The variant selection threshold was set at 100% of the halo diameter of the PAM reference when the median value of the distribution was lower than 100% but was set at the median value when this latter value was higher than 100% (positions S101 and S103). Detail of the number of selected variants per position is indicated.
