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.

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 method for enzymatic sulfurylation of a substrate is provided which includes the steps of reacting the substrate with 3′-phosphoadenosine-5′-phosphosulfate (PAPS) in a medium containing a bacterium belonging to the family Enterobacteriaceae to produce a sulfated derivative of the substrate, and collecting the sulfated derivative from the medium, wherein the bacterium has been modified to produce, at least, a protein having sulfotransferase activity, and to attenuate expression of an aphA gene, a cysQ gene, or a cpdB gene, or a combination of these.

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 method of treating a subject suffering from a neurodegenerative disease characterized by insufficient autophagy is provided, the method comprising administering to the subject an effective amount of a composition that inhibits N-deacetylase N-sulfotransferase (NDST), Further provided is a method of identifying a modulator of autophagy.

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.

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 method for enzymatic sulfurylation of a substrate is provided which includes the steps of reacting the substrate with 3-phosphoadenosine-5-phosphosulfate (PAPS) in a medium containing a bacterium belonging to the family Enterobacteriaceae to produce a sulfated derivative of the substrate, and collecting the sulfated derivative from the medium, wherein the bacterium has been modified to produce, at least, a protein having sulfotransferase activity, and to attenuate expression of an aphA gene, a cysQ gene, or a cpdB gene, or a combination of these.

The disclosure relates to double stranded ribonucleic acid (dsRNAi) agents and compositions targeting a heparan sulfate biosynthesis pathway enzyme gene (HSBPE) gene, e.g., Exostosin Glycosyltransferase 1 (EXT1), Exostosin Glycosyltransferase 2 (EXT2), and/or N-Deacetylase And N-Sulfotransferase 2, (NDST2 gene), as well as methods of inhibiting expression of an HSBPE gene and methods of treating subjects having Mucopolysaccaridosis type III (MPS III), e.g., MPS IIIA, MPS IIIB, MPS IIIC, or MPS IIID, using such dsRNAi agents and compositions.

An anticoagulant heparin-chondroitin chimeric saccharide molecule as well as a preparation method and application thereof are disclosed. The anticoagulant heparin-chondroitin chimeric saccharide molecule has a structure as shown in formula I. The heparin-chondroitin chimeric saccharide molecule of the present disclosure has potent activities against an Xa factor and IIa, and the activity of the heparin-chondroitin chimeric saccharide molecule can be effectively neutralized by protamine, with a neutralization rate of greater than or equal to 70%. The risk of causing adverse reactions such as fatal HIT is obviously lower than that of enoxaparin and other low-molecular-weight heparins. The heparin-chondroitin chimeric saccharide molecule disclosed by the present disclosure is suitable for the preparation of a safer potent anticoagulant and antithrombotic new drug.