38 results about "Endoglycosidase" patented technology

A chemoenzymatic method for the preparation of a core-fucoslyated glycoprotein or glycopeptide, including (a) providing an acceptor selected from the group consisting of a fucosylated GlcNAc-protein and fucosylated GlcNAc-peptide; and (b) reacting the acceptor with a donor substrate including an activated oligosaccharide moiety, in the presence of an endoglycosidase (ENGase) selected from Endo;F1, Endo-F2, Endo-F3, Endo-D and related glycosynthase mutants to transfer the oligosaccharide moiety to the acceptor and yield the structure defined core-fucosylated glycoprotein or glycopeptide. The donor substrate includes, in a specific implementation, a synthetic oligosaccharide oxazoline. A related method of fucosylated glycoprotein or fucosylated glycopeptide remodeling with a predetermined natural N-glycan or a tailor-made oligosaccharide moiety, and a method of remodeling an antibody to include a predetermined sugar chain to replace a heterogeneous sugar chain, are also described.

The invention relates to a method for analyzing protein O-glycosylation sites. The method comprises the following steps: (1), sequentially carrying out endonuclease digestion, endoglycosidase and exoglycosidase digestion on a proteome sample or a single protein sample to prepare a peptide and non-glycopeptide protein sample or single protein sample with a GalNAc label; (2) loading the sample on a durian agglutinin chromatographic column and separating to prepare a proteomic sample; (3) separating the single protein sample or the proteomic sample through with a C18 reversed-phase chromatographic column, and then detecting by a high-definition mass spectrometer in a cation mode to obtain a high-definition mass spectrogram; (4) processing the obtained mass spectrometric data and obtaining protein glycosylation site information though the searched result. The site information can be more rapidly, more accurately and more comprehensively obtained through establishing the method for detecting the O-glycosylation sites of glycoproteins in the single protein sample and the proteomic sample.

The invention provides use of an EndoS polypeptide, or a polynucleotide encoding an EndoS polypeptide, in the manufacture of a medicament for the treatment or prevention of a disease or condition mediated by IgG antibodies.

The present invention relates to inhibition of heparinoids anti-coagulation activity by a non-active form of a eukaryotic endoglycosidase or any fragment or peptide thereof comprising at least one heparin-binding domain. More particularly, the invention provides compositions and methods for the inhibition of heparinoids anti-coagulation activity and for the treatment of coagulation related pathologic clinical conditions, using a non-active form of mammalian heparanase or peptides thereof comprising at least one heparin-binding domain.

The invention belongs to the technical field of analytical chemistry, and particularly relates to a method for catalyzing an isotope labeled N-sugar chain by using endoglycosidase. Glycoprotein enzymolysis reaction of the endoglycosidase is performed in 18O water, and when the N-sugar chain is digested by enzyme, an isotope 18O molecule is labeled at the reductive tail end of the N-sugar chain. Mass spectrometric detection results prove that the difference between the isotope 18O labeled sugar chain molecular weight and the unlabeled sugar chain molecular weight is 2 Daltons and the isotope 18O molecule is stably labeled at the reductive tail end of the sugar chain. The method is simple and efficient, fills the blank of the sugar chain isotope 18O labeling technology, and can be effectively used for relative quantification of the sugar chain.

The invention discloses a preparation method of egg source protein powder with high characteristics and the protein powder prepared by using the preparation method. The preparation method comprise thefollowing steps: (1) selecting egg white liquid and uniformly stirring; (2) taking a proper amount of egg white liquid, adjusting the pH value to be 5.0-6.0, adding dry yeast and glucose, and insulating in a water bath kettle at the temperature of 45 DEG C to obtain an activated seed liquid; adjusting the pH value of the residual egg white liquid to be 5.0-6.0, adding activated seed liquid, and fermenting for 2-3 h in a water bath kettle at the temperature of 30 DEG C; (3) adding 500-1000 U/g of neutral protease and papain at the ratio of 1: (1-3), carrying out enzymolysis on the neutral protease and the papain for 60-90 min, and adding endoglycosidase when the enzymolysis reaches 40 min; (4) carrying out activated carbon decolorization to obtain decolorized enzymatic hydrolysate; and (5)adding vegetable protein, mogroside, beta-dextran, hydroxypropyl starch and lecithin; and then carrying out electric field induction treatment for 4-7 minutes, and carrying out spray drying to obtainthe egg source protein powder; and the egg source protein powder prepared by the invention has the characteristics of immune activity, high digestibility and absorption, balanced nutrition, no bittertaste and excellent taste, and is a brewing instant terminal product.

Genetically engineered host animal cells capable of producing glycoproteins having modified glycosylation patterns, e.g., defucosylation and/or monoglycosylation. Such host animal cells can be engineered to express fucosidase, endoglycosidase or both.

A mutant of Endos2 includes one or more mutations in the sequence of a wild-type EndoS2 (SEQ ID NO:1), wherein the one or more mutations are in a peptide region located within residues 133-143, residues 177-182, residues 184-189, residues 221-231, and/or residues 227-237, wherein the mutant of EndoS2 has a low hydrolyzing activity and a high tranglycosylation activity, as compared to those of the wild-type EndoS2. A method for preparing an engineered glycoprotein using the mutant of EndoS2 includes coupling an activated oligosaccharide to a glycoprotein acceptor. The activated oligosaccharide is a glycan oxazoline.

The invention relates to a method for realizing the engineering expression of glycosyl endoglycosidase EGCase I. The method can realize the effective soluble expression of the EGCase I and maintain excellent hydrolysis and transglycosylation activity; a novel EGCase I mutant enzyme developed on the basis of the method has a broad substrate spectrum, and also has the activity of glycosidase synthetase; and the obtained glycosphingolipid endoglycosidase is very suitable for being applied to the analysis and the synthesis of industrial glycosphingolipids.