Abstract: In one aspect, methods of biomolecular analysis are described herein. Briefly, a method comprises providing a composition comprising glycosaminoglycans and contacting the composition with a membrane comprising at least one nanopore. An electric field is applied across the nanopore, and data of glycosaminoglycan translocation events through the nanopore are recorded. A molecular weight distribution of the glycosaminoglycans is derived from the data.

Abstract: In one aspect, methods of biomolecular analysis are described herein. Briefly, a method comprises providing a composition comprising glycosaminoglycans and contacting the composition with a membrane comprising at least one nanopore. An electric field is applied across the nanopore, and data of glycosaminoglycan translocation events through the nanopore are recorded. A molecular weight distribution of the glycosaminoglycans is derived from the data.

Abstract: The present invention relates to Factor IX polypeptides conjugated to heparosan (HEP) polymers, methods for the manufacture thereof and uses of such conjugates. The resultant conjugates may be used—for example—in the treatment or prevention of bleeding disorders such as haemophilia B.

Abstract: The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Pasteurella multocida hyaluronate synthase (PmHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product.

Abstract: The present invention relates to a recombinantly produced, enzymatically active hyaluronan synthase, wherein the recombinantly produced, enzymatically active hyaluronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcA and UDP-GlcNAc to synthesize HA. The recombinantly produced, enzymatically active hyaluronan synthase may be produced from a hyaluronan synthase gene isolated from a microbial source, such as a streptococcal source.

Abstract: The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate or chondroitin or heparin/heparosan synthases from Pasteurella, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure with a targeted size that are substantially monodisperse in size. The present invention also relates to methodology for polymer grafting by a polysachharide synthase to form glycosaminoglycan polymers having an unnatural structure.

Abstract: The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate or chondroitin or heparin/heparosan synthases from Pasteurella, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure with a targeted size that are substantially monodisperse in size. The present invention also relates to methodology for polymer grafting by a polysaccharide synthase to form glycosaminoglycan polymers having an unnatural structure.

Abstract: The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Pasteurella multocida hyaluronate synthase (PmHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product.

Abstract: The present invention relates to a Gram-negative glycosaminoglycan gene and methods of making and using same. The present invention relates to recombinant Gram-positive host cells containing a Gram-negative glycosaminoglycan synthase gene, and methods of producing glycosaminoglycans using such recombinant host cells.

Abstract: The present invention relates to a recombinant Bacillus host cell containing a recombinant vector including a nucleic acid segment having a coding region segment encoding enzymatically active hyaluronan synthase (HAS). The recombinant Bacillus host cell is utilized in a method for producing hyaluronic acid (HA).

Abstract: The present invention relates to a nucleic acid segment having a coding region segment encoding enzymatically active Pasteurella multocida hyaluronate synthase (PmHAS), and to the use of this nucleic acid segment in the preparation of recombinant cells which produce hyaluronate synthase and its hyaluronic acid product.

Abstract: The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate or chondroitin or heparin/heparosan synthases from Pasteurella, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure with a targeted size that are substantially monodisperse in size.

Abstract: The present invention relates to methodology for polymer grafting by a polysaccharide synthase and, more particularly, polymer grafting using the hyaluronate or chondroitin or heparin/heparosan synthases from Pasteurella multocida, in order to create a variety of glycosaminoglycan oligosaccharides having a natural or chimeric or hybrid sugar structure.

Abstract: The present invention relates to a recombinantly produced, enzymatically active hyaluronan synthase, wherein the recombinantly produced, enzymatically active hyaluronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcA and UDP-GlcNAc to synthesize HA. The recombinantly produced, enzymatically active hyaluronan synthase may be produced from a hyaluronan synthase gene isolated from a microbial source, such as a streptococcal source.

Abstract: A functionally active hyaluronan synthase having at least one modified amino acid residue therein as compared to a corresponding functionally active native hyaluronan synthase such that the functionally active hyaluronan synthase has an altered enzymatic activity as compared to the corresponding functionally active native hyaluronan synthase is disclosed. The altered enzymatic activity may result in increased or decreased activity when compared to the corresponding native hyaluronan synthase, or the altered enzymatic activity may result in production of hyaluronan having an average molecular mass that is greater than or less than an average molecular mass of hyaluronan produced by the corresponding native hyaluronan synthase. Methods of producing hyaluronic acid utilizing a recombinant host cell having an expression construct encoding the functionally active hyaluronan synthase with altered enzymatic activity are also disclosed.

Abstract: The present invention relates to a method of producing hyaluronic acid that includes introducing a purified nucleic acid segment having a coding region encoding enzymatically active hyaluronan synthase into a Bacillus strain, and culturing the Bacillus strain under conditions appropriate for the production of hyaluronic acid.

Abstract: The present invention relates to a recombinantly produced, enzymatically active hyaluronan synthase, wherein the recombinantly produced, enzymatically active hyaluronan synthase is a single protein that is a dual-action catalyst that utilizes UDP-GlcA and UDP-GlcNAc to synthesize HA. The recombinantly produced, enzymatically active hyaluronan synthase may be produced from a hyaluronan synthase gene isolated from a microbial source, such as a streptococcal source.

Abstract: Disclosed are DNA sequences encoding hyaluronic acid synthase that are employed to construct recombinant cells useful in the production of hyaluronata synthase and hyaluronic acid (HA). In preferred aspects, chromosomal DNA encoding the HA synthase gene, hasA, was cloned from a Streptococcus pyoganas genomic library. These vectors were used to transform host cells such as E. coli and acapsular Streptococci to produce hyaluronic acid. Resultant transformants were screened to identify colonies which have incorporated HA synthase DNA in a form that is being actively transcribed into the corresponding HA synthase enzyme. These colonies were selected and employed in the production of hyaluronic acid.

Abstract: The present invention relates to a recombinant host cell that includes an expression construct having a hyaluronan synthase gene that encodes an enzymatically active hyaluronan synthase (HAS), and to the use of these recombinant cells that produce hyaluronan synthase and its hyaluronic acid product.

Abstract: The present invention relates to a recombinant Bacillus host cell containing a recombinant vector including a nucleic acid segment having a coding region segment encoding enzymatically active hyaluronan synthase (HAS). The recombinant Bacillus host cell is utilized in a method for producing hyaluronic acid (HA).