Production in yeast of α-galactosidase A, a lysosomal enzyme applicable to enzyme replacement therapy for Fabry disease

Yasunori Chiba, Hitoshi Sakuraba, Masaharu Kotani, Ryoichi Kase, Kazuo Kobayashi, Makoto Takeuchi, Satoshi Ogasawara, Yutaka Maruyama, Tasuku Nakajima, Yuki Takaoka, Yoshifumi Jigami

Research output: Contribution to journalReview articlepeer-review

49 Citations (Scopus)


A mammalian-like sugar moiety was created in glycoprotein by Saccharomyces cerevisiae in combination with bacterial α-mannosidase to produce a more economic enzyme replacement therapy for patients with Fabry disease. We introduced the human α-galactosidase A (α-GalA) gene into an S. cerevisiae mutant that was deficient in the outer chains of N-linked mannan. The recombinant α-GalA contained both neutral (Man8GlcNAc2) and acidic ([Man-P]1-2Man8GlcNAc2) sugar chains. Because an efficient incorporation of α-GalA into lysosomes of human cells requires mannose-6-phosphate (Man-6-P) residues that should be recognized by the specific receptor, we trimmed down the sugar chains of the α-GalA by a newly isolated bacterial α-mannosidase. Treatment of the α-GalA with the α-mannosidase resulted in the exposure of a Man-6-P residue on a nonreduced end of oligosaccharide chains after the removal of phosphodiester-linked nonreduced-end mannose. The treated α-GalA was efficiently incorporated into fibroblasts derived from patients with Fabry disease. The uptake was three to four times higher than that of the nontreated α-GalA and was inhibited by the addition of 5 mM Man-6-P. Incorporated α-GalA was targeted to the lysosome, and hydrolyzed ceramide trihexoside accumulated in the Fabry fibroblasts after 5 days. This method provides an effective and economic therapy for many lysosomal disorders, including Fabry disease.

Original languageEnglish
Pages (from-to)821-828
Number of pages8
Issue number12
Publication statusPublished - 2002 Dec 1
Externally publishedYes


  • Lysosomal disease
  • Mannose-6-phosphate
  • Mannosidase
  • Saccharomyces cerevisiae
  • Therapeutic glycoprotein

ASJC Scopus subject areas

  • Biochemistry


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