title: Sequence analysis and resistance to pepsin hydrolysis as part of an assessment of the potential allergenicity of ice structuring protein type III HPLC 12.
abstract: The recently published WHO/FAO guidelines on the assessment of allergenicity of novel food proteins provide a strategy with which to approach the determination of the potential of novel proteins in foods to be allergens. Key to this strategy are the assessment of sequence similarity to known allergens and the assessment of the resistance to pepsin hydrolysis. Ice structuring proteins (also commonly referred to as anti-freeze or thermal hysteresis proteins) are a group of naturally occurring proteins that bind to ice and structure ice crystal formation. The amino acid sequence of the ice structuring protein (ISP) type III HPLC 12 (ISP type III) was compared in silico with the sequences of known allergens. Secondly, the resistance to pepsin hydrolysis of ISP type III and its glycoconjugates (produced in recombinant baker's yeast) was assessed. The results indicate that ISP type III has no sequence similarity with known allergenic proteins. Both ISP type III and ISP type III glycoconjugates contained within the fermentation product were hydrolysed readily by pepsin (50% loss in <10 min at pH 1.5) to give peptide fragments that were too small to be allergenic or to trigger cross-linking to IgE. In an accompanying study, we demonstrated that IgE from fish-allergic individuals did not bind ISP Type III. Therefore, in accordance with the WHO/FAO strategy, the assessment of ISP type III and ISP type III glycoconjugates by sequence analysis together with lack of resistance to pepsin hydrolysis and the absence of IgE binding supports the conclusion that both are unlikely to present a potential sensitisation hazard.
authors.affiliation: Safety and Environmental Assurance Centre, Unilever Colworth Laboratory, Colworth House, Sharnbrook, Bedfordshire MK44 1LQ, UK.
authors.forename: B
authors.initials: B
authors.lastname: Baderschneider
authors.forename: R W R
authors.initials: RW
authors.lastname: Crevel
authors.forename: L K
authors.initials: LK
authors.lastname: Earl
authors.forename: A
authors.initials: A
authors.lastname: Lalljie
authors.forename: D J
authors.initials: DJ
authors.lastname: Sanders
authors.forename: I J
authors.initials: IJ
authors.lastname: Sanders
mesh_terms: D000485:Allergens / Q000009:adverse effects* / Q000737:chemistry / Q000378:metabolism; D002851:Chromatography, High Pressure Liquid; D004591:Electrophoresis, Polyacrylamide Gel; D006868:Hydrolysis; D013058:Mass Spectrometry / Q000379:methods; D010434:Pepsin A / Q000378:metabolism*
chemical_list: D000485:Allergens; D010434:Pepsin A