Lactococcus lactis as a vehicle for the heterologous expression of fungal ribotoxin variants with reduced IgE-binding affinity

Fungal ribotoxins are a family of extracellular ribonucleases which inhibit protein biosynthesis by inactivating the ribosomes. This inactivation results in the induction of cell death by apoptosis. Ribotoxins show antitumoral properties based on their ability to cross the membrane of some transform...

Full description

Saved in:
Bibliographic Details
Published in:Journal of biotechnology 2008-03, Vol.134 (1), p.1-8
Main Authors: Álvarez-García, Elisa, Alegre-Cebollada, Jorge, Batanero, Eva, Monedero, Vicente, Pérez-Martínez, Gaspar, García-Fernández, Rosa, Gavilanes, Jose G., Martínez del Pozo, Álvaro
Format: Article
Language:English
Subjects:
Animals
Asp f 1
Aspergillus giganteus
Biological and medical sciences
Biotechnology
Electrophoresis, Polyacrylamide Gel
Endoribonucleases - genetics
Endoribonucleases - metabolism
Female
Fundamental and applied biological sciences. Psychology
Fungal Proteins - genetics
Fungal Proteins - metabolism
Genetic Engineering - methods
Immunoglobulin E - metabolism
Intestine, Small - microbiology
Lactococcus lactis
Lactococcus lactis - genetics
Lactococcus lactis - metabolism
Mice
Mice, Inbred BALB C
Mycotoxins - genetics
Mycotoxins - metabolism
Plasmids - genetics
Probiotic
Protein Binding
Ribonuclease
α-Sarcin
Citations: Items that this one cites
Items that cite this one
Online Access:Get full text
Tags: Add Tag
No Tags, Be the first to tag this record!
Description
Summary:Fungal ribotoxins are a family of extracellular ribonucleases which inhibit protein biosynthesis by inactivating the ribosomes. This inactivation results in the induction of cell death by apoptosis. Ribotoxins show antitumoral properties based on their ability to cross the membrane of some transformed cells. Unfortunately, they also show an unspecific cytotoxicity which has greatly impaired their potential clinical uses. α-Sarcin, produced by Aspergillus giganteus, is the best-characterized ribotoxin. Asp f 1, another ribotoxin produced by A. fumigatus, is indeed one of its major allergens. In this work, the Lactococcus lactis MG1363 strain has been engineered to produce and secrete not only wild-type Asp f 1 and α-sarcin but also three different mutants with reduced cytotoxicity and/or IgE-binding affinity. The proteins were secreted in native and active form when the extracellular medium employed was buffered at pH values around 8.0. Strains producing the wild-type natural α-sarcin were proved to be innocuous when administered intragastrically to mice for a period of 14 days. Overall, the results presented are discussed in terms of its potential application as a vehicle of oral delivery of hypoallergenic variants as well as a starting point to approach the design of strategies to accomplish the safe delivery of these proteins as antitumoral agents.
ISSN:0168-1656
1873-4863
DOI:10.1016/j.jbiotec.2007.12.011