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Stable coexpression of two human sialylation enzymes in plant suspension-cultured tobacco cells

Paper ID Volume ID Publish Year Pages File Format Full-Text
20900 43196 2011 7 PDF Available
Title
Stable coexpression of two human sialylation enzymes in plant suspension-cultured tobacco cells
Abstract

Human CMP-N-acetylneuraminic acid (NeuAc) synthase (hCSS) and α2,6-sialyltransferase (hST) participate in the sialylation of N-linked glycans in mammalian cells. hCSS synthesizes CMP-NeuAc, which hST uses as a donor substrate to transfer NeuAc to the terminal position of N-linked glycans. In plant cells, the presence of NeuAc has not yet been substantiated and the identification of the genes involved in the sialylation of N-glycan has not been carried out. In this study, we introduced hCSS and hST genes into suspension-cultured tobacco BY2 cells to provide the machinery for the sialylation pathway in plants. hCSS and hST stably expressed in the plant cells showed activity. In addition, CMP-NeuAc produced by hCSS in the transformed plant cells functioned as a donor substrate to hST. An in vitro coupled hCSS and hST reaction resulted in the production of mammalian-type sialoglycoproteins bearing terminal NeuAc residues. Furthermore, the results of the purification of the coupled-reaction products by Sambucus sieboldian lectin column chromatography and digestion with linkage-specific neuraminidase revealed that the modified terminal residue was α2,6-linked NeuAc. Here, we demonstrate that the in vitro sialylation of N-linked glycans on mammalian proteins can be achieved using plant cell extracts stably expressing hCSS and hST, providing proof-of-principle that a sialylated human therapeutic protein can be produced in plants.

Keywords
Sialic acid; N-glycan; Plant suspension-cultured cell; CMP-N-acetylneuraminic acid synthase; Sialyltransferase
First Page Preview
Stable coexpression of two human sialylation enzymes in plant suspension-cultured tobacco cells
Publisher
Database: Elsevier - ScienceDirect
Journal: Journal of Bioscience and Bioengineering - Volume 111, Issue 4, April 2011, Pages 471–477
Authors
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Subjects
Physical Sciences and Engineering Chemical Engineering Bioengineering