Stable coexpression of two human sialylation enzymes in plant suspension-cultured tobacco cells(PLANT BIOTECHNOLOGY)

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概要

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• 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.

• 公益社団法人日本生物工学会の論文

著者

• Fujiyama Kazuhito

  International Center for Biotechnology, Osaka University

• MISAKI Ryo

  International Center for Biotechnology, Osaka University

• SEKI Tatsuji

  International Center for Biotechnology, Osaka University

• Kazuhito Fujiyama

  International Center For Biotechnology Osaka University

• Fujiyama Kazuhito

  大阪大学生物工学国際交流センター

• Fujiyama Kazuhito

  International Center For Biotechnology Osaka Univ.

• Fujiyama Kazuhito

  Department Of Fermentation Technology Osaka University

• Fujiyama Kazuhito

  International Center For Biotechnology Osaka University

• Misaki Ryo

  International Center For Biotechnology Osaka University

• Misaki Ryo

  The International Center For Biotechnology Osaka University

• Seki Tatsuji

  International Center For Biotechnology Osaka University

• Seki Tatsuji

  大阪大学生物工学国際交流センター

• Fujiyama Kazuhito

  The International Center For Biotechnology Osaka Univ. 2-1 Yamada-oka Suita Osaka 565-0871 Jpn

• Kajiura Hiroyuki

  International Center for Biotechnology, Osaka University

• Kajiura Hiroyuki

  International Center For Biotechnology Osaka University

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