Methodologies for labeling the epigenetic modification 5-hydroxymethyl-cytosine (5hmC) along a DNA molecule, and for determining a presence or a level of this epigenetic modification based on a ratio of fluorescence intensity of a labeled DNA sample to absorption intensity of the DNA sample at 260 nm are disclosed. Related compositions and reagents, and methods of preparing same are also disclosed.

The present invention provides a recombinant microorganism for producing citicoline and a method for producing citicoline by using the recombinant microorganism, wherein genes for degradation and utilization of citicoline, choline, and phosphocholine are knocked out, In addition, a pyrimidine nucleoside synthesis pathway is genetically engineered to remove feedback inhibition to the synthesis pathway. A yield of more than 20 g/L of citicoline can be obtained with recombinant strains in a 5-liter fermenter by means of a biological fermentation method, achieving industrial mass production with low citicoline production costs and less pollution; therefore, the method is a simple, environmentally friendly and has a relatively high promotion and application value.

The present invention relates to an enzyme-catalyzed process for producing UDP-galactose from low-cost substrates uridine monophosphate and D-galactose in a single reaction mixture. The process can be operated (semi)continuously or in batch mode. The process can be extended to uridine as starting material instead of uridine monophosphate. Further, the process can be adapted to produce galactosylated molecules and biomolecules including saccharides, proteins, peptides, glycoproteins or glycopeptides, particularly human milk oligosaccharides (HMO) and (monoclonal) antibodies.

The present invention relates to an enzyme-catalyzed process for producing UDP-N-acetyl-α-D-glucosamine (UDP-GlcNAc) from low-cost substrates uridine monophosphate and N-acetyl-D glucosamine in a single reaction mixture with immobilized or preferably co-immobilized enzymes. Uridine may be used as starting material instead of uridine monophosphate as well. Further, the process may be adapted to produce GlcNAcylated molecules and biomolecules including saccharides, particularly human milk oligosaccharides (HMO), proteins, peptides, glycoproteins, particularly antibodies, or glycopeptides, and bioconjugates, particularly carbohydrate conjugate vaccines and antibody-drug conjugates.

A method or producing amino sugar (containing) products using metabolically engineered microorganisms is disclosed, wherein the conversion of UDP-N-acetylglucosamine to cell envelope precursors and molecules is reduced by altering the activity of enzymes involved in the synthesis of cell envelope precursors and molecules.

Disclosed herein are oligosaccharides and intermediates useful for the production thereof. The compounds are useful as analytical standards and as intermediates for the preparation of more complex oligosaccharide and N-glycan products. The compounds may be prepared in high purity using the selective stop/go synthetic methods disclosed herein.

The present invention relates to a modified (mutant) nicotinamide phosphoribosyltransferase, and a method for producing nicotinamide mononucleotide using the same. The modified nicotinamide phosphoribosyltransferase (Nampt) of the present invention comprises the amino acid sequence represented by the following SEQ ID NO: 1 and/or the amino acid sequence represented by the following SEQ ID NO: 2, and has improved activity as compared with wild-type Nampt: (a) SEQ ID NO: 1: S- [V/I] -P-A-X.sub.1-X.sub.2-H-S-[T/V/I] - [M/V/I] -X.sub.3, and (b) SEQ ID NO: 2: X.sub.4-[S/I] -D-X.sub.5 wherein X.sub.1 to X.sub.5 are as defined herein.

The present invention relates to a modified (mutant) nicotinamide phosphoribosyltransferase, and a method for producing nicotinamide mononucleotide using the same. The modified nicotinamide phosphoribosyltransferase (Nampt) of the present invention comprises the amino acid sequence represented by the following SEQ ID NO: 1 and/or the amino acid sequence represented by the following SEQ ID NO: 2, and has improved activity as compared with wild-type Nampt: (a) SEQ ID NO: 1: S- [V/I] -P-A-X.sub.1-X.sub.2-H-S-[T/V/I] - [M/V/I] -X.sub.3, and (b) SEQ ID NO: 2: X.sub.4-[S/I] -D-X.sub.5 wherein X.sub.1 to X.sub.5 are as defined herein.

The present invention provides circularized RNA and methods of making, purifying, and using same.

The present invention provides circularized RNA and methods of making, purifying, and using same.