University of Arkansas

Other

Neose Technologies, Inc.

Alan D Elbein

Gary A Bannon

Richard Drake

Y T Pan

Richard R Drake

Catherine M Ketcham

Irena Pastuszak

Irena Patuszak

Sam Smith

The present invention provides a purified, homogeneous plant enzyme that adds a &#x3b2;-1,2-linked xylose to the &#x3b2;-linked mannose on the N-linked oligosaccharides of storage glycoproteins. This &#x3b2;1,2-xylosyltransferase was purified from the microsomal fraction of soybean cells approximately 51,000-fold. Also provided is an antibody recognizing this &#x3b2;1,2-xylosyltransferase.

Antibody directed against purified &#x3b2; 1,2-xylosyltransferase

The present invention provides a purified, homogeneous plant enzyme that adds a &bgr;-1,2-linked xylose to the &bgr;-linked mannose on the N-linked oligosaccharides of storage glycoproteins. This &bgr;1,2-xylosyltransferase was purified from the microsomal fraction of soybean cells approximately 51,000-fold. Also provided is polyclonal antiserum recognizing this &bgr;1,2-xylosyltransferase enzyme and uses thereof.

Purified &bgr;1,2-xylosyltransferase and uses thereof

A new enzyme that phosphorylates GalNAc at position 1 to form GalNAc-&agr;-1P was purified &tilde;1275-fold from the cytosolic fraction of pig kidney, and the properties of the enzyme were determined. The kinase is specific for GalNAc as the phosphate acceptor and is inactive with GlcNAc, ManNAc, glucose, galactose, mannose, GalN, and GlcN. The native enzyme has a molecular mass of 48-51 kDa, and this enzyme is clearly separated from galactokinase by chromatography. The GalNAc kinase has a pH optimum between 8.5 and 9.0, and requires a divalent cation in the order Mg2&plus;>Mn2&plus;>Co2&plus;, with optimum Mg2&plus; concentration at &tilde;5 mM. The enzyme was most active with ATP as the phosphate donor, but slight activity was observed with ITP, acetyl-P, and phosphoenolpyruvate. Enzyme activity was highest in porcine and human kidney and porcine liver, and was low in most other tissues.

Liver GalNAc-1-phosphate kinase

The present invention provides a purified, homogeneous plant enzyme that adds a &bgr;-1,2-linked xylose to the &bgr;-linked mannose on the N-linked oligosaccharides of storage glycoproteins. This &bgr;1,2-xylosyltransferase was purified from the microsomal fraction of soybean cells approximately 51,000-fold. Also provided is polyclonal antiserum recognizing this &dgr;1,2-xylosyltransferase enzyme and uses thereof.

The present invention provides an enzyme L-fucokinase isolated from porcine kidney and in homogeneously purified form. The enzyme has a native molecular weight of 440 kDa based on gel filtration, a subunit molecular weight of about 110 kDa based on SDS PAGE, an optimal pH of about 8.0, and wherein said enzyme catalyzes the phosphorylation of L-fucose and does not phosphorylate D-glucose, D-galactose and D-mannose.

Purified porcine kidney L-fucose kinase

DNA sequences encoding GDP-fucose pyrophosphorylase (GTP+fucose-1-P.fwdarw.GDP-fucose+PP.sub.i), are provided. The enzymes can be used in the synthesis of desired carbohydrate structures.

Nucleic acids encoding GDP-Fucose pyrophosphorylase

The present invention describes a method for the treatment of gram-negative bacterial infections using high-mannose containing oligosaccharides. Specifically the use of Man.sub.9 (GlcNAc).sub.2 -hydrophobic glycopeptides (i.e. tyrosinamide) to block adhesion of the bacteria pili to the oligosaccharide of the host cells plasma membrane in infections of Enterobacter cloacae and other Enterobacter and gram-negative species.

Inhibition of bacterial binding by high-mannose oligosaccharides

The present invention provides the enzyme UDP-N-acetylglucosamine pyrophosphorylase in isolated and purified form. Also provided is a various method of using and preparing this purified, homogeneous enzyme.

Growing community of inventors

Little Rock, AR, United States of America

Alan D Elbein