UDP-galactose: β-N-acetyl-glucosamine β-1,4-galactosyltransferase, β4Gal-T2

US6916649B2 · kind B2 · utility

Assignee

• GlycoZym ApS · DK

Inventors

• Henrik Clausen · Seattle, US
• Eric Paul Bennett · Avenue Madeleine, DK

Key dates

• Filing date: Apr 24, 2002
• Grant date: Jul 12, 2005
• Expiry date: Mar 26, 2023

Classification

• Technology area (CPC C): Chemistry; Metallurgy
• CPC primary: C12N9/1051
• WIPO field: Biotechnology
• WIPO sector: Chemistry

Abstract

A novel gene defining a novel enzyme in the UDP-D-galactose: b-N-acetyl-glucosamine β-1,4-galactosyltransferase family, termed β4Gal-T2, with unique enzymatic properties is disclosed. The enzymatic activity of β4Gal-T2 is shown to be distinct from that of previously identified enzymes of this gene family. The invention discloses isolated DNA molecules and DNA constructs encoding β4Gal-T2 and derivatives thereof by way of amino acid deletion, substitution or insertion exhibiting β4Gal-T2 activity, as well as cloning and expression vectors including such DNA, cells transfected with the vectors, and recombinant methods for providing β4Gal-T2. The enzyme β4Gal-T2 and β4Gal-T2-active derivatives thereof are disclosed, in particular soluble derivatives comprising the catalytically active domain of β4Gal-T2. Further, the invention discloses methods of obtaining β-1,4-galactosyl glycosylated saccharides, glycopeptides or glycoproteins by use of an enzymically active β4Gal-T2 protein or fusion protein thereof or by using cells stably transfected with a vector including DNA encoding an enzymatically active β4Gal-T2 protein as an expression system for recombinant production of such glycopepti…