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Characterization and Gene Cloning of an Acidic Thaumatin-Like Protein (TLP 1), an Allergen from Sapodilla Fruit (Manilkara zapota)

Ashok Kumar, Hassan G and Venkatesh, L. Hegde and Santoshkumar, M. Shetty and Venkatesh, Y. P. (2013) Characterization and Gene Cloning of an Acidic Thaumatin-Like Protein (TLP 1), an Allergen from Sapodilla Fruit (Manilkara zapota). Allergology International, 62 (4). pp. 447-462.

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Abstract

Background: Allergy to sapodilla (Manilkara zapota) fruit ingestion is rare. An independent study from our group has identified a basic thaumatin-like protein (TLP 2) as the major allergen. The present study was aimed at identifying and characterizing additional allergens from sapodilla. Methods: Allergic subjects were identified by case history, skin prick test (SPT) and allergen-specific IgE. Sapodilla extract was fractionated using SP-Sepharose into 3 components (SP1, SP2 and SP3) which were analyzed by native�SDS-PAGE, IgE-immunoblot, isoelectric focusing (IEF) and N-terminal sequencing. The conserved regions of plant TLPs and the N-terminal sequence were used to design primers for PCR. Results: SPT and ELISA confirmed a subject with oral allergy syndrome (OAS) to sapodilla and custard apple. Two proteins (26.9 and 24.5 kDa; reducing conditions) were detected as allergens, of which the latter in SP2 has already been identified as basic TLP (TLP 2). The 26.9 kDa protein present in SP1 was identified as an acidic TLP based on native PAGE, IEF and N-terminal sequencing. Presence of a basic β-1,3-glucanase in SP3 was inferred by zymography. Sequence analysis of the genomic clone of the acidic TLP gene revealed that it is intronless and non-glycosylated. Evolutionary relatedness to olive, grape and kiwi fruit allergenic TLPs were inferred by phylogenetic analysis. Conclusions: An acidic TLP (TLP 1) was identified as a new allergen in sapodilla. TLP 1 is a single polypeptide (207 residues) belonging to the thaumatin family of the GH64-TLP-SF superfamily. Clinically, sapodilla should be considered in the list of fruits causing OAS.

Item Type: Article
Uncontrolled Keywords: acidic TLP, exotic fruit, food allergen, Manilkara zapota, oral allergy syndrome
Subjects: 600 Technology > 08 Food technology > 12 Allergy & Toxicology
600 Technology > 08 Food technology > 24 Fruits
Divisions: Dept. of Biochemistry
Depositing User: Food Sci. & Technol. Information Services
Date Deposited: 20 May 2016 10:27
Last Modified: 20 May 2016 10:27
URI: http://ir.cftri.com/id/eprint/12195

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