@article{01HSXMHFFZVXMWSYMKPJ5GEYPX,
  abstract     = {{The reconstruction of ancestral sequences can offer a glimpse into the fascinating process of molecular evolution by exposing the adaptive pathways that shape the proteins found in nature today. Here, we track the evolution of the carbohydrate-active enzymes responsible for the synthesis and turnover of mannogen, a critical carbohydrate reserve in Leishmania parasites. Biochemical characterization of resurrected enzymes demonstrated that mannoside phosphorylase activity emerged in an ancestral bacterial mannosyltransferase, and later disappeared in the process of horizontal gene transfer and gene duplication in Leishmania. By shuffling through plausible historical sequence space in an ancestral mannosyltransferase, we found that mannoside phosphorylase activity could be toggled on through various combinations of mutations at positions outside of the active site. Molecular dynamics simulations showed that such mutations can affect loop rigidity and shield the active site from water molecules that disrupt key interactions, allowing alpha-mannose 1-phosphate to adopt a catalytically productive conformation. These findings highlight the importance of subtle distal mutations in protein evolution and suggest that the vast collection of natural glycosyltransferases may be a promising source of engineering templates for the design of tailored phosphorylases.}},
  author       = {{Franceus, Jorick and  Rivas-Fernandez, Jose Pablo and Lormans, Jolien and  Rovira, Carme and Desmet, Tom}},
  issn         = {{2155-5435}},
  journal      = {{ACS CATALYSIS}},
  keywords     = {{enzyme engineering,enzyme evolution,ancestralsequence reconstruction,glycoside phosphorylase,GT108,Leishmania,mannogen,CENTRAL CARBON METABOLISM,MOLECULAR-DYNAMICS,FAMILY,HYDROLASES,MECHANISM,VIRULENCE}},
  language     = {{eng}},
  number       = {{5}},
  pages        = {{3103--3114}},
  title        = {{Evolution of phosphorylase activity in an ancestral glycosyltransferase}},
  url          = {{http://doi.org/10.1021/acscatal.3c05819}},
  volume       = {{14}},
  year         = {{2024}},
}

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