Recombinant Allergens Production in Yeast.
The methylotropic yeast Pichia pastoris has been extensively used in large-scale production of properly folded recombinant proteins. As an eukaryotic organism P. pastoris presents a series of advantages at expression and processing of heterologous proteins such as post-translational modifications, protein processing, and a reasonably sophisticated quality control of protein folding when compared against Escherichia coli. In this chapter, we describe the modified lab procedure for cloning and expression in Pichia pastoris of common food allergens sequences from the raw fruit to the fully folded biotinylated protein product.
School of Biosciences, University of Nottingham, College Road, Sutton Bonington Campus, Sutton Bonington, Loughborough, LE12 5RD, UK. maria.neophytou@nottingham.ac.uk.
Maria
M
Neophytou
School of Biosciences, University of Nottingham, Sutton Bonington Campus, Leicestershire, UK.
Marcos
M
Alcocer
D000485:Allergens / Q000235:genetics*; D004926:Escherichia coli / Q000235:genetics; D005512:Food Hypersensitivity / Q000235:genetics; D010843:Pichia / Q000235:genetics*; D017510:Protein Folding; D011499:Protein Processing, Post-Translational / Q000235:genetics; D011994:Recombinant Proteins / Q000235:genetics*
D000485:Allergens; D011994:Recombinant Proteins
Food allergens; In vivo biotinylation; Pichia pastoris; Protein expression; Protein tagging; Yeast expression