Glycan engineering of proteins with whole living yeast cells expressing rat liver α2,3‐sialytransferase in the porous cell wall

FEBS Letters

Heikinheimo, Riikka¹  Makarow, Marja¹  Sievi, Eeva¹  Helin, Jari¹ 

[1] Institute of Biotechnology, P.O. Box 56, University of Helsinki, 00014 Helsinki, Finland

关键词: Sialyltransferase;    Yeast;    Glycan engineering;    Recombinant pharmaceutical;    Secretion;   

DOI  :  10.1016/S0014-5793(98)01550-6

学科分类：生物化学/生物物理

来源: John Wiley & Sons Ltd.

【 摘 要 】

The N-glycans of recombinant proteins produced via the secretory pathway of cultured mammalian cells are often undersialylated, and insect cells lack sialytransferases. Undersialylated glycoproteins are rapidly cleared from the circulation, compromising the effect of pharmaceuticals. We show that incubation with living Saccharomyces cerevisiae cells expressing the catalytic ectodomain of rat liver α2,3-sialyltransferase (ST3N_e) in the porous cell wall resulted in sialylation of glycoproteins. The K _m values of the yeast enzyme for several substrates were similar to those of recombinant ST3N_e from insect cells and of authentic ST3N. The yeast strain provides an inexpensive self-perpetuating source of ST3N activity for glycan engineering of recombinant proteins.

【 授权许可】

Unknown