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US 8,137,928 B2Granted
Expression of soluble, active eukaryotic glycosyltransferases in prokaryotic organisms
AssigneeBiogenerix AG
InventorsMarc F. Schwartz, Tarik Soliman
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Abstract
The present invention provides enhanced methods of producing soluble, active eukaryotic glycosyltransferases in prokaryotic microorganisms that have an oxidizing environment.
Claims5
A method of producing a soluble truncated active eukaryotic glycosyltransferase in a prokaryotic microorganism, which method comprising the steps of
- (a) providing a prokaryotic microorganism that has an oxidizing intracellular environment,
- (b) expressing a nucleic acid that encodes a truncated eukaryotic glycosyltransferase in the prokaryotic microorganism, wherein the truncated eukaryotic glycosyltransferase is a truncated human GalNacT2 of SEQ ID NO:
- 33 having a truncation selected from the group consisting of Δ51 Δ445, or one or two truncations selected from the group consisting of Δ40, Δ53, Δ73, Δ94, and Δ445, and
- (c) growing the microorganism under conditions that allow expression of the soluble truncated active eukaryotic glycosyltransferase within a cellular compartment of the prokaryotic microorganism.
1 more element
A method of producing a soluble active eukaryotic glycosyltransferase fusion protein in a prokaryotic microorganism, which method comprises:
- (a) providing a prokaryotic microorganism that has an oxidizing intracellular environment,
- (b) expressing a nucleic acid that encodes a soluble active eukaryotic glycosyltransferase fusion protein which comprises a maltose binding protein (MBP) and a truncated eukaryotic glycosyltransferase, wherein the truncated eukaryotic glycosyltransferase is a truncated human GalNAcT2 of SEQ ID NO:
- 33 having a truncation selected from the group consisting of Δ51Δ445, or one or two truncations selected from the group consisting of Δ40, Δ53, Δ73, Δ94, and Δ445, in the prokaryotic microorganism, and
- (c) growing the microorganism under conditions that allow expression of the soluble active eukaryotic glycosyltransferase fusion protein within a cellular compartment of the prokaryotic microorganism.
1 more element
Dependencies are inferred from the claim text.