A bacterial expression system revisited for the recombinant production of cystine-rich plant lipid transfer proteins.
Non-specific lipid transfer proteins (nsLTPs) are abundant and ubiquitous cystine-rich proteins that are capable, in vitro, of binding lipids and hydrophobic molecules. In view to probe the lipid binding properties of the wheat nsLTP1, mutant variants may represent a powerful tool. To this end, a synthetic gene, encoding a mature wheat nsLTP1 polypeptide, was designed to ensure high level expression in Escherichia coli. The bacterial expression host strain, a translational fusion strategy, and convenient cleavage and purification procedures were optimized to produce in standard fermentation conditions, a significant amount (15 mg/L final yield), of a soluble and correctly folded recombinant nsLTP1. This highly amenable expression system is helpful in order to investigate structure-activity relationships of plant nsLTP.
Elmorjani
Khalil
K
Unité de Recherche sur les Protéines Végétales et leurs Interactions, INRA, Rue de la Géraudière 44316 Nantes Cedex 3, France. elmorjan@nantes.inra.fr
Lurquin
Vanessa
V
Lelion
André
A
Rogniaux
Hélène
H
Marion
Didier
D
D052179:Antigens, Plant; D002352:Carrier Proteins / Q000096:biosynthesis* / Q000737:chemistry / Q000235:genetics*; D003553:Cystine / Q000737:chemistry / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000235:genetics* / Q000378:metabolism*; D015964:Gene Expression Regulation, Bacterial / Q000235:genetics; D024861:Genetic Enhancement / Q000379:methods*; D016297:Mutagenesis, Site-Directed; D010940:Plant Proteins; D015202:Protein Engineering / Q000379:methods*; D011994:Recombinant Proteins / Q000096:biosynthesis; D014908:Triticum / Q000235:genetics* / Q000378:metabolism*
D052179:Antigens, Plant; D002352:Carrier Proteins; D010940:Plant Proteins; D011994:Recombinant Proteins; C511920:lipid transfer proteins, plant; D003553:Cystine