US 2014/0154707 A1 · N-GLYCOSYLATION IN TRANSFORMED PHAEODACTYLUM TRICORNUTUM

N-glycosylation in transformed phaeodactylum tricornutum

AssigneesUniversite De Rouen, L'institut Francais De Recherche Pour L'exploitation De La Mer (Ifremer), Algenics

InventorsMuriel Bardor, Romain Louvet, Bruno Saint-Jean, Carole Burel, Berangere Baiet, Jean-Paul Cadoret, Patrice Lerouge, Remy Michel, Aude Carlier

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Abstract

Transformed Phaeodactylum tricornutum including a nucleic acid sequence operatively linked to a promoter, wherein the nucleic acid sequence encodes an N-acetylglucosaminyltransferase I and/or an α-Mannosidase II and wherein at least one β-N-acetylglucosaminidase of the transformed Phaeodactylum tricornutum has been inactivated. A method for producing a glycosylated polypeptide includes the steps of (i) culturing a transformed P. tricornutum as defined previously and (ii) purifying the polypeptide that is expressed and glycosylated in the transformed P. Tricornutum. The use of such a transformed P. tricornutum for producing a glycosylated polypeptide is also described.

Claims23

Dependent claims

1-15. (canceled)

A transformed Phaeodactylum tricornutum whose N-glycosylation pathway has been modified by the inactivation of at least one β-N-acetylglucosaminidase and/or the expression of at least one glycosylation enzyme encoded by a nucleic acid sequence operatively linked to a promoter, wherein

(i) said nucleic acid sequence encodes an N-acetylglucosaminyltransferase I having the amino acid sequence SEQ ID No 1, a fragment or a derivative thereof and said operatively linked promoter has a sequence identity of less 50% with SEQ ID No 3 or a fragment thereof, and/or
(ii) said nucleic acid sequence encodes an α-Mannosidase II having the amino acid sequence SEQ ID No 5, a fragment or a derivative thereof and said operatively linked promoter has a sequence identity of less 50% with SEQ ID No 7 or a fragment thereof.

Dependencies are inferred from the claim text.