title: High levels of stable phytase accumulate in the culture medium of transgenic Medicago truncatula cell suspension cultures.
abstract: The use of plants for production of recombinant proteins is becoming widely accepted. More recently, plant cell cultures have been proposed as valuable systems for producing a wide range of biologically active proteins. Such systems provide certain advantages over whole plants, but yields are still considered a limitation. In this study we established a Medicago truncatula cell suspension line expressing phytase from Aspergillus niger. Phytase is an N-glycosylated enzyme that breaks down indigestible phytate, resulting in an increased availability of phosphorus and other minerals in monogastric animals and reduced levels of phosphorus output in their manure. Various production systems have previously been used to express heterologous phytase, including several plant species. In this work, remarkable amounts of enzymatically active recombinant phytase were produced and secreted into the culture medium. Recombinant phytase accumulated to at least 25 mg/L and remained stable along the growth curve, and an enriched fraction with high enzymatic activity was easily obtained. We therefore propose M. truncatula cell suspension cultures as a potential system for the production of recombinant proteins. Most importantly, we have shown that, contrary to general belief, it is possible to achieve high levels of a functional recombinant protein in plant cell culture systems.
authors.affiliation: Plant Cell Biology Laboratory, Instituto de Tecnologia Quimica e Biologica, UNL, Oeiras, Portugal.
authors.forename: Ana Sofia
authors.initials: AS
authors.lastname: Pires
authors.forename: M Guadalupe
authors.initials: MG
authors.lastname: Cabral
authors.forename: Pedro
authors.initials: P
authors.lastname: Fevereiro
authors.forename: Eva
authors.initials: E
authors.lastname: Stoger
authors.forename: Rita
authors.initials: R
authors.lastname: Abranches
mesh_terms: D010832:6-Phytase / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism; D001234:Aspergillus niger / Q000201:enzymology; D002478:Cells, Cultured; D003470:Culture Media; D004591:Electrophoresis, Polyacrylamide Gel; D005656:Fungal Proteins / Q000096:biosynthesis; D018506:Gene Expression Regulation, Plant; D046913:Medicago truncatula / Q000201:enzymology* / Q000235:genetics* / Q000254:growth & development / Q000378:metabolism; D010833:Phytic Acid / Q000378:metabolism; D030821:Plants, Genetically Modified; D011994:Recombinant Proteins / Q000096:biosynthesis* / Q000302:isolation & purification / Q000378:metabolism
chemical_list: D003470:Culture Media; D005656:Fungal Proteins; D011994:Recombinant Proteins; D010833:Phytic Acid; D010832:6-Phytase