Provided herein are genetically modified yeast cells capable of producing one or more human milk oligosaccharides. The yeast cells include one or more heterologous nucleic acids that encode enzymes of a human milk oligosaccharide biosynthetic pathway. The yeast cells do not include a heterologous nucleic acid encoding a fucokinase. Also provided are fermentation compositions including the disclosed genetically modified yeast cells, and related methods of producing and recovering human milk oligosaccharides generated by the yeast cells.

The present invention provides a novel biosynthetic production process which converts L-galactose into 2-fucosyllactose via four enzymatically catalyzed reaction steps. The present process is designed such that co-factors required by the process are regenerated within the four reaction steps, hence making the process cost-effective and efficient. The process can be performed in vitro in a cell-free system. The present invention also provides mutant enzymes that can be used to increase production levels of 2-fucosyllactose, whether using the novel pathway described herein or the mannose-dependent pathway known in the art.

Microbial cells genetically engineered with a heterologous nucleic acid sequence that increases export of 2 fucosyllactose are disclosed. Methods of increasing export of 2 fucosyllactose from a microbial cell and for identifying a heterologous nucleic acid sequence that increases export of 2 fucosyllactose from a microbial cell are also disclosed.

The present invention relates to a method for producing fucosylated oligosaccharides by using a recombinant prokaryotic host cell that is cultivated on a gluconeogenic substrate, as well as to the host cell and its use. The host cell is genetically modified in that the activity of a fructose-6-phosphate converting enzyme is abolished or lowered, and the transport of the produced fucosylated oligosaccharide through the cell membrane is facilitated by an exogenous transport protein.

The present invention relates to an enzyme-catalyzed process for producing GDP-fucose from low-cost substrates guanosine and L-fucose or guanosine and D-Mannose in a single reaction mixture. Said process can be operated (semi)continuously or in batch mode. Further, said process can be adapted to produce fucosylated molecules and biomolecules including glycans, such as human milk oligosaccharides, proteins, peptides, glycoproteins or glycopeptides.

Microbial cells genetically engineered with a heterologous nucleic acid sequence that increases export of 2 fucosyllactose are disclosed. Methods of increasing export of 2 fucosyllactose from a microbial cell and for identifying a heterologous nucleic acid sequence that increases export of 2 fucosyllactose from a microbial cell are also disclosed.

The present invention relates to a method for producing fucosylated oligosaccharides by using a recombinant prokaryotic host cell that is cultivated on a gluconeogenic substrate, as well as to the host cell and its use. The host cell is genetically modified in that the activity of a fructose-6-phosphate converting enzyme is abolished or lowered, and the transport of the produced fucosylated oligosaccharide through the cell membrane is facilitated by an exogenous transport protein.

The present invention relates to a method for producing fucosylated oligosaccharides by using a recombinant prokaryotic host cell that is cultivated on a gluconeogenic substrate, as well as to the host cell and its use. The host cell is genetically modified in that the activity of a fructose-6-phosphate converting enzyme is abolished or lowered, and the transport of the produced fucosylated oligosaccharide through the cell membrane is facilitated by an exogenous transport protein.

The present invention is in the technical field of synthetic biology and metabolic engineering. More particularly, the present invention is in the technical field of metabolically engineered cells and use of said cell in a cultivation, preferably a fermentation. The present invention describes a cell for the production of a compound. The cell comprises a pathway for the production of the compound, which can be a disaccharide, oligosaccharide and/or a Neu(n)Ac-containing bioproduct, wherein (n) is 4, 5, 7, 8 or 9 or a combination thereof. The cell is metabolically engineered for enhanced synthesis of acetyl-Coenzyme A. The invention also resides in a method of producing such compound by cultivation, preferably a fermentation, with such a cell.

The present disclosure relates to a method of obtaining a cell where fucosylation pathways are modified, leading to production of partially fucosylated and non-fucosylated protein products, specifically antibodies from the cell. The present disclosure employs the Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR) technology. The method of the present disclosure targets the Fut8 gene and GMD gene in a cell. Such products are used in developing therapeutics and biomarkers, and in diagnosis and prognosis of diseases.