Fig. 2: Ancestral LPMO reconstruction and characterization.
From: Enzymatic upgrading of nanochitin using an ancient lytic polysaccharide monooxygenase
a Uncorrelated lognormal relaxed-clock chronogram of LPMO with geological time inferred with Bayesian inference. A total of fifty-one LPMO homologous sequences were used. Geological scale and times are indicated in the upper bar. The selected internal node for experimental validation is shown with a circle LFACA (Last Firmicutes Actinobacteria Common Ancestor). b LFACA-LPMO (red) and BtLPMO (blue) AlphaFold2 prediction structure alignment with RMSD value of 0.428. c Active site Cu ion (yellow) and neighboring interacting residues at approximately 5 Å distance, conforming the so-called “histidine-brace”. d 2,6-DMP activity assay for LFACA-LPMO (red squares) and BtLPMO (blue dots) at different temperatures, 30 °C to 90 °C. e pH activity assay from pH 3 to pH 10, LFACA-LPMO (red squares) and BtLPMO (blue dots). For each data point, six independent experiments were collected. Data are shown as average ± S.E.M. f Generation of nanochitin over time by oxidative cleavage of α-Chitin by LFACA-LPMO (red squares) and extant BtLPMO (blue dots). For each data point, three independent experiments were collected. Data are shown as average ± S.D.
