Human α-mannosidase produced in transgenic tobacco plants is processed in human α-mannosidosis cell lines
Article first published online: 7 JUN 2011
© 2011 The Authors. Plant Biotechnology Journal © 2011 Society for Experimental Biology, Association of Applied Biologists and Blackwell Publishing Ltd
Plant Biotechnology Journal
Volume 9, Issue 9, pages 1061–1073, December 2011
How to Cite
De Marchis, F., Balducci, C., Pompa, A., Riise Stensland, H. M. F., Guaragno, M., Pagiotti, R., Menghini, A. R., Persichetti, E., Beccari, T. and Bellucci, M. (2011), Human α-mannosidase produced in transgenic tobacco plants is processed in human α-mannosidosis cell lines. Plant Biotechnology Journal, 9: 1061–1073. doi: 10.1111/j.1467-7652.2011.00630.x
- Issue published online: 9 NOV 2011
- Article first published online: 7 JUN 2011
- Received 4 October 2010; revised 31 March 2011; accepted 31 March 2011.
- transgenic tobacco;
- vacuolar targeting;
- enzyme replacement therapy
Deficiency in human lysosomal α-mannosidase (MAN2B1) results in α-mannosidosis, a lysosomal storage disorder; patients present a wide range of neurological, immunological, and skeletal symptoms caused by a multisystemic accumulation of mannose-containing oligosaccharides. Here, we describe the expression of recombinant MAN2B1 both transiently in Nicotiana benthamiana leaves and in the leaves and seeds of stably transformed N. tabacum plants. After purification from tobacco leaves, the recombinant enzyme was found to be N-glycosylated and localized in vacuolar compartments. In the fresh leaves of tobacco transformants, MAN2B1 was measured at 10 200 units/kg, and the purified enzyme from these leaves had a specific activity of 32–45 U/mg. Furthermore, tobacco-produced MAN2B1 was biochemically similar to the enzyme purified from human tissues, and it was internalized and processed by α-mannosidosis fibroblast cells. These results strongly indicate that plants can be considered a promising expression system for the production of recombinant MAN2B1 for use in enzyme replacement therapy.