Human α-mannosidase produced in transgenic tobacco plants is processed in human α-mannosidosis cell lines.
Plant Biotechnol J
; 9(9): 1061-73, 2011 Dec.
Article
em En
| MEDLINE
| ID: mdl-21645202
ABSTRACT
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.
Texto completo:
1
Coleções:
01-internacional
Base de dados:
MEDLINE
Assunto principal:
Nicotiana
/
Alfa-Manosidase
Limite:
Humans
Idioma:
En
Revista:
Plant Biotechnol J
Assunto da revista:
BIOTECNOLOGIA
/
BOTANICA
Ano de publicação:
2011
Tipo de documento:
Article
País de afiliação:
Itália