The present invention relates to a process for the enzymatic modification of a glycoprotein. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of glycosyltransferase that is, or is derived from, a ?-(1,4)-N-acetylgalactosaminyltransferase, with a non-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotide is according to formula (3):

##STR00001##

wherein A is selected from the group consisting of N.sub.3, C(O)R.sup.3, (CH.sub.2).sub.iC?CR.sup.4, SH, SC(O)R.sup.8, SC(O)OR.sup.8, SC(S)OR.sup.8, F, Cl, Br I, OS(O).sub.2R.sup.5, terminal C.sub.2-C.sub.24 alkenyl groups, C.sub.3-C.sub.5 cycloalkenyl groups, C.sub.4-C.sub.8 alkadienyl groups, terminal C.sub.3-C.sub.24 allenyl groups and amino groups. The invention further relates to a glycoprotein obtainable by the process according to the invention, to a bioconjugate that can be obtained by conjugating the glycoprotein with a linker-conjugate, and to ?-(1,4)-N-acetylgalactosaminyltransferases that can be used in preparing the glycoprotein according to the invention.

The present invention relates to a process for the enzymatic modification of a glycoprotein. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of glycosyltransferase that is, or is derived from, a ?-(1,4)-N-acetylgalactosaminyltransferase, with a non-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotide is according to formula (3): wherein A is selected from the group consisting of N.sub.3, C(O)R.sup.3, (CH.sub.2).sub.iC?CR.sup.4, SH, SC(O)R.sup.8, SC(O)OR8, SC(S)OR8, F, CI, Br I, OS(O).sub.2R.sup.5, terminal C.sub.2-C.sub.24 alkenyl groups, C.sub.3-C.sub.5 cycloalkenyl groups, C.sub.4-C.sub.8 alkadienyl groups, terminal C.sub.3-C.sub.24 allenyl groups and amino groups. The invention further relates to a glycoprotein obtainable by the process according to the invention, to a bioconjugate that can be obtained by conjugating the glycoprotein with a linker-conjugate, and to ?-(1,4)-N-acetylgalactosaminyltransferases that can be used in preparing the the glycoprotein according to the invention. ##STR00001##

Provided herein are host cells capable of producing a human milk oligosaccharide (HMO), such as yeast cells that are deficient in expression or activity of an endogenous oxidoreductase. Also provided are fermentation compositions including the disclosed host cells, as well as related methods of producing and recovering HMOs generated by the host cells.

The disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, the disclosure is in the technical field of metabolically engineered cells and use of the cells in a cultivation or fermentation. The disclosure describes a cell and a method for production of a di- and/or oligosaccharide. The cell comprises a pathway for production of the di- and/or oligosaccharide and is genetically modified for expression and/or overexpression of at least one set of multiple coding DNA sequences wherein the multiple coding DNA sequences within one set differ in nucleotide sequence and each encode a polypeptide, wherein the polypeptides have the same function and/or activity of interest. Furthermore, the disclosure provides for purification of the di- and/or oligosaccharide from the cultivation.

This technology relates to the production of lactose and human milk oligosaccharides (HMOs) in cells.

The present invention relates to a process for the enzymatic modification of a glycoprotein. The process comprises the step of contacting a glycoprotein comprising a glycan comprising a terminal GlcNAc-moiety, in the presence of glycosyltransferase that is, or is derived from, a -(1,4)-N-acetylgalactosaminyltransferase, with a non-natural sugar-derivative nucleotide. The non-natural sugar-derivative nucleotide is according to formula (3): wherein A is selected from the group consisting of N.sub.3, C(O)R.sup.3, (CH.sub.2).sub.iCCR.sup.4, SH, SC(O)R.sup.8, SC(0)OR8, SC(S)OR8, F, CI, Br I, OS(O).sub.2R.sup.5, terminal C.sub.2-C.sub.24 alkenyl groups, C.sub.3-C.sub.5 cycloalkenyl groups, C.sub.4-C.sub.8 alkadienyl groups, terminal C.sub.3-C.sub.24 allenyl groups and amino groups. The invention further relates to a glycoprotein obtainable by the process according to the invention, to a bioconjugate that can be obtained by conjugating the glycoprotein with a linker-conjugate, and to -(1,4)-N-acetylgalactosaminyltransferases that can be used in preparing the the glycoprotein according to the invention.

##STR00001##

The present invention provides means and methods for the production in eukaryotic cells of homogeneous forms of small glycan structures which carry terminal galactose residues. In addition, the invention provides glycan-conjugates based on specific coupling with galactose residues present on the recombinant glycoproteins.

This invention relates to a method of producing one or more human milk oligosaccharides (HMOs), in particular LNT and/or LNnT, in a genetically engineered cell comprising an enhanced oligosaccharide transport capability. The genetically modified cell comprises a series of genetic modification which enable the production of one or more HMO(s), and a series of genetic modification that enhances the transport of lactose and produced HMO(s).

Disclosed are genetically engineered microbial cells for the production of oligosaccharides comprising a galactose-1,4-glucose moiety at their reducing end, wherein said microbial cells are able to produce said oligosaccharides in the absence of exogenously added lactose, and a method of producing said oligosaccharides using said microbial cells.

The present disclosure relates to improved strains for the production of Human Milk Oligosaccharides (HMOs) in large scale. The strains are genetically engineered to produce less acetate and/or ethanol during large-scale fermentation, in particular when encountering gradients with excess carbon source in the fermenter.