Journal
TALANTA
Volume 240, Issue -, Pages -Publisher
ELSEVIER
DOI: 10.1016/j.talanta.2021.123188
Keywords
Food allergens; Proteomics; Spirulina; Chlorella; Microalgae; FTMS; Tandem MS
Categories
Funding
- Italian Ministero per l'Istruzione, l'Universit`a e la Ricerca (MIUR) [PONa3_00395/1, 2017YER72K]
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With the introduction of novel nutrient sources with high protein content in the consumer market, safety evaluation studies on potentially allergenic proteins are necessary. This study presents a method for establishing sequence-based homology between proteins of spirulina and chlorella micro-algae and known allergens through database searches. Several potentially allergenic proteins were identified, and further immunochemical tests are required to confirm their allergenicity.
Since novel nutrient sources with high protein content, such as yeast, fungi, bacteria, algae, and insects, are increasingly introduced in the consumer market, safety evaluation studies on their potentially allergenic proteins are required. A pipeline for in silico establishing the sequence-based homology between proteins of spirulina (Arthrospira platensis) and chlorella (Chlorella vulgaris) micro-algae and those included in the AllergenOnline (AO) database (AllergenOnline.org) is described. The extracted proteins were first identified through tryptic peptides analysis by reversed-phase liquid chromatography and high resolution/accuracy Fourier-transform tandem mass spectrometry (RPLC-ESI-FTMS/MS), followed by a quest on the UniProt database. The AO database was subsequently interrogated to assess sequence similarity between identified microalgal proteins and known allergens, based on criteria established by the World Health Organization (WHO) and Food and Agriculture Organization (FAO). A direct search for microalgal proteins already included in allergen databases was also performed using the Allergome database. Six proteins exhibiting a significant homology with food allergens were identified in spirulina extracts. Five of them, i.e., two thioredoxins (D4ZSU6, K1VP15), a superoxide dismutase (C3V3P3), a glyceraldehyde-3-phosphate dehydrogenase (K1W168), and a triosephosphate isomerase (D5A635), resulted from the search on AO. The sixth protein, C-phycocyanin beta subunit (P72508), was directly obtained after examining the Allergome database. Two proteins exhibiting significant sequence homology with food allergens were retrieved in chlorella extracts, viz. calmodulin (A0A2P6TFR8), which is related to troponin c (D7F1Q2), and fructose-bisphosphate aldolase (A0A2P6TDD0). Specific serum screenings based on immunochemical tests should be undertaken to confirm or rule out the allergenicity of the identified proteins.
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