Open AccessHuman α‐mannosidase produced in transgenic tobacco plants is processed in human α‐mannosidosis cell lines
Author(s) -
De Marchis Francesca,
Balducci Chiara,
Pompa Andrea,
Riise Stensland Hilde M. F.,
Guaragno Marco,
Pagiotti Rita,
Menghini Anna R.,
Persichetti Emanuele,
Beccari Tommaso,
Bellucci Michele
Publication year - 2011
Publication title -
plant biotechnology journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 3.525
H-Index - 115
eISSN - 1467-7652
pISSN - 1467-7644
DOI - 10.1111/j.1467-7652.2011.00630.x
Subject(s) - nicotiana tabacum , nicotiana benthamiana , mannosidase , biology , recombinant dna , enzyme , transgene , mannose , solanaceae , enzyme assay , biochemistry , cell culture , genetically modified crops , gene , genetics
Summary 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.