Department of Chemistry, University of Bari Aldo Moro, Via Orabona 4, 70126, Bari, Italy.
Department of Chemistry, University of Bari Aldo Moro, Via Orabona 4, 70126, Bari, Italy.
Talanta. 2022 Apr 1;240:123188. doi: 10.1016/j.talanta.2021.123188. Epub 2021 Dec 30.
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.
由于具有高蛋白含量的新型营养源，如酵母、真菌、细菌、藻类和昆虫，越来越多地被引入消费市场，因此需要对其潜在过敏原蛋白进行安全性评估研究。本文描述了一种基于序列同源性的计算方法，用于建立螺旋藻（Arthrospira platensis）和小球藻（Chlorella vulgaris）微藻的蛋白质与过敏原在线（AllergenOnline，AO）数据库（AllergenOnline.org）中包含的蛋白质之间的序列同源性。通过反相高效液相色谱和高分辨率/高精度傅里叶变换串联质谱（RPLC-ESI-FTMS/MS）对提取的蛋白质进行胰蛋白酶肽分析，然后在 UniProt 数据库中进行检索。随后，根据世界卫生组织（WHO）和粮食及农业组织（FAO）制定的标准，对 AO 数据库进行查询，以评估鉴定出的微藻蛋白与已知过敏原之间的序列相似性。还使用 Allergome 数据库对已包含在过敏原数据库中的微藻蛋白进行了直接搜索。在螺旋藻提取物中鉴定出 6 种与食物过敏原具有显著同源性的蛋白质。其中 5 种，即 2 种硫氧还蛋白（D4ZSU6，K1VP15）、1 种超氧化物歧化酶（C3V3P3）、1 种甘油醛-3-磷酸脱氢酶（K1W168）和 1 种磷酸丙糖异构酶（D5A635），是通过对 AO 的搜索得到的。第 6 种蛋白质，C-藻蓝蛋白β亚基（P72508），是在检查 Allergome 数据库后直接获得的。在小球藻提取物中也检索到 2 种与食物过敏原具有显著序列同源性的蛋白质，即钙调蛋白（A0A2P6TFR8），它与肌钙蛋白 C（D7F1Q2）有关，和果糖-1,6-二磷酸醛缩酶（A0A2P6TDD0）。应进行基于免疫化学测试的特异性血清筛查，以确认或排除鉴定出的蛋白质的致敏性。
