The present invention relates a polypeptide having kaurenoic acid 13-hydroxylase activity, which polypeptide comprises an amino acid sequence which, when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1 or SEQ ID NO: 3, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 136, 248, 336 or 403, said positions being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

The present invention relates a polypeptide having kaurenoic acid 13-hydroxylase activity, which polypeptide comprises an amino acid sequence which, when aligned with a kaurenoic acid 13-hydroxylase comprising the sequence set out in SEQ ID NO: 1 or SEQ ID NO: 3, comprises at least one substitution of an amino acid corresponding to any of amino acids at positions 136, 248, 336 or 403, said positions being defined with reference to SEQ ID NO: 1 or SEQ ID NO: 3 and wherein the polypeptide has one or more modified properties as compared with a reference polypeptide having kaurenoic acid 13-hydroxylase activity. A polypeptide of the invention may be used in a recombinant host for the production of steviol or a steviol glycoside.

A method for producing allolactose is provided. Allolactose is produced by bringing cells of a microorganism having beta-galactosidase (BGL) into contact with lactose in the presence of a substance that inhibits hydrolysis of allolactose.

This invention relates to a method for the enzymatic synthesis of oligosaccharides, preferably human milk oligosaccharides (HMOs) The method comprises the enzymatic transfer of a glycosyl moiety and subsequent removal of by-products, such as lactose, by nanofiltration using a membrane comprising an active polyamide layer.

This invention relates to a method for the enzymatic synthesis of oligosaccharides, preferably human milk oligosaccharides (HMOs) The method comprises the enzymatic transfer of a glycosyl moiety and subsequent removal of by-products, such as lactose, by nanofiltration using a membrane comprising an active polyamide layer.

The present invention provides a new type of galacto-oligosaccharide, having a mannose residue instead of a glucose residue at the reducing end. The invention also relates to compositions comprising this galacto-oligosaccharide, its preparation and use in nutritional compositions.

The present invention provides a new type of galacto-oligosaccharide, having a mannose residue instead of a glucose residue at the reducing end. The invention also relates to compositions comprising this galacto-oligosaccharide, its preparation and use in nutritional compositions.

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 metabolically engineered cell and a method by cultivation or fermentation with the cell for production of a sialylated di- and/or oligosaccharide. The metabolically engineered cell comprises a pathway for production of the sialylated di- and/or oligosaccharide and is modified for expression and/or overexpression of multiple coding DNA sequences encoding one or more isoproteins that catalyze the same chemical reaction. Furthermore, the disclosure provides for purification of the sialylated di- and/or oligosaccharide from the cultivation.

The disclosure is in the technical field of synthetic biology and metabolic engineering. More particularly, the disclosure is in the technical field of cultivation or fermentation of metabolically engineered cells. The disclosure describes a method for the production of a di- or oligosaccharide with an N-acetylglucosamine at the reducing end by a cell as well as the purification of the di- or oligosaccharide from the cultivation. Furthermore, the disclosure provides a cell metabolically engineered for production of a di- or oligosaccharide with an N-acetylglucosamine at the reducing end.

Clean-in-place methods applicable to improve production of fermentation operations are disclosed herein to monitor, report, and control the cleaning reagents applied, residual components thereof, and clean-in-place conditions within levels that are compatible with fermentation processes, without limiting the effectiveness of the clean-in-place protocol with respect to cleaning or sanitizing the fermentation system.