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US 11,773,382 B2Granted
Engineered aryl sulfate-dependent enzymes
AssigneeOptimvia, LLC
InventorsTarsis Gesteira Ferreira, Daniel H. Lajiness
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AssigneeOptimvia, LLC
InventorsTarsis Gesteira Ferreira, Daniel H. Lajiness
Estimating the term…
The present invention provides several non-naturally occurring sulfotransferase enzymes that have been engineered to react with aryl sulfate compounds as sulfo group donors, instead of the natural substrate 3′-phosphoadenosine 5′-phosphosulfate (PAPS), and with heparosan-based polysaccharides, particularly heparan sulfate, as sulfo group acceptors. Each of the engineered sulfotransferase enzymes have a biological activity characterized by the position within the heparosan-based polysaccharide that receives the sulfo group, including glucosaminyl N-sulfotransferase activity, hexuronyl 2-O sulfotransferase activity, glucosaminyl 6-O sulfotransferase activity, or glucosaminyl 3-O sulfotransferase activity. Methods of using the engineered sulfotransferases to produce sulfated heparosan-based polysaccharides, including polysaccharides having anticoagulant activity, are also provided.
A non-natural hexuronyl 2-O sulfotransferase (2OST) enzyme having an amino acid sequence comprising multiple mutations relative to conserved amino acid residues and sequence motifs found in natural 2OST enzymes within enzyme class EC 2.8.2.-, wherein:
A non-natural hexuronyl 2-O sulfotransferase (2OST) enzyme engineered to have sulfotransferase activity in the absence of 3′-phosphoadenosine 5′-phosphosulfate (PAPS), the sulfotransferase activity comprising a transfer of a sulfo group from an aryl sulfate compound to a heparosan-based polysaccharide to form a 2-O-sulfated heparan sulfate product, wherein the heparan sulfate comprises N-sulfated heparan sulfate, and the 2-O -sulfated heparan sulfate product comprises N-,2-O-sulfated heparan sulfate,
Dependencies are inferred from the claim text.