Expanding the Substrate Scope of a Bacterial Nucleotidyltransferase via Allosteric Mutations

Maggie Zheng, Meng Zheng, Tania J. Lupoli

Research output: Contribution to journalArticlepeer-review


Bacterial glycoconjugates, such as cell surface polysaccharides and glycoproteins, play important roles in cellular interactions and survival. Enzymes called nucleotidyltransferases use sugar-1-phosphates and nucleoside triphosphates (NTPs) to produce nucleoside diphosphate sugars (NDP-sugars), which serve as building blocks for most glycoconjugates. Research spanning several decades has shown that some bacterial nucleotidyltransferases have broad substrate tolerance and can be exploited to produce a variety of NDP-sugars in vitro. While these enzymes are known to be allosterically regulated by NDP-sugars and their fragments, much work has focused on the effect of active site mutations alone. Here, we show that rational mutations in the allosteric site of the nucleotidyltransferase RmlA lead to expanded substrate tolerance and improvements in catalytic activity that can be explained by subtle changes in quaternary structure and interactions with ligands. These observations will help inform future studies on the directed biosynthesis of diverse bacterial NDP-sugars and downstream glycoconjugates.

Original languageEnglish (US)
Pages (from-to)2035-2044
Number of pages10
JournalACS Infectious Diseases
Issue number10
StatePublished - Oct 14 2022


  • NDP-sugars
  • RmlA
  • allosteric regulation
  • bacterial glycoconjugates
  • bacterial nucleotidyltransferase
  • mutational analysis

ASJC Scopus subject areas

  • Infectious Diseases


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