Romarís-Hortas V, Bianga J, Moreda-Piñeiro A, Bermejo-Barrera P, Szpunar J
Department of Analytical Chemistry, Nutrition and Bromatology, Faculty of Chemistry, University of Santiago de Compostela, Avenida das Ciencias, s/n. 15782 - Santiago de Compostela, Spain.
CNRS/UPPA, Laboratoire de Chimie Analytique Bio-inorganique et Environnement, UMR5254, Hélioparc, 2, Av. Angot, 64053 Pau, France.
Talanta. 2014 Sep;127:175-80. doi: 10.1016/j.talanta.2014.04.003. Epub 2014 Apr 13.
An analytical approach providing an insight into speciation of iodine in water insoluble fraction of edible seaweed (Nori) was developed. The seaweed, harvested in the Galician coast (Northwestern Spain), contained 67.7±1.3 μg g(-1) iodine of which 25% was water soluble and could be identifies as iodide. Extraction conditions of water insoluble residue using urea, NaOH, SDS and Triton X-100 were investigated. The protein pellets obtained in optimized conditions (after precipitation of urea extracts with acetone), were digested with trypsin and protease XIV. Size exclusion chromatography-ICP-MS of both enzymatic digests demonstrated the occurrence of iodoaminoacids putatively present in proteins. Intact proteins could be separated by gel electrophoresis after an additional extraction of the protein extract with phenol. Sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS PAGE) with laser ablation ICP-MS detection of (127)I indicated the presence of iodine in protein bands corresponding to molecular masses of 110 kDa, 40 kDa, 27 kDa, 20 kDa and 10 kDa. 2D IEF-SDS PAGE with laser ablation ICP-MS (127)I imaging allowed the detection of 5 iodine containing protein spots in the alkaline pI range.
开发了一种分析方法,用于深入了解可食用海藻(紫菜)水不溶部分中碘的形态。这种在加利西亚海岸(西班牙西北部)收获的海藻含有67.7±1.3μg g⁻¹的碘,其中25%是水溶性的,可鉴定为碘化物。研究了使用尿素、氢氧化钠、十二烷基硫酸钠(SDS)和聚乙二醇辛基苯基醚(Triton X-100)对水不溶残渣的提取条件。在优化条件下(用丙酮沉淀尿素提取物后)获得的蛋白质沉淀,用胰蛋白酶和蛋白酶XIV进行消化。两种酶解产物的尺寸排阻色谱-电感耦合等离子体质谱(SEC-ICP-MS)表明蛋白质中可能存在碘代氨基酸。在用苯酚对蛋白质提取物进行额外提取后,完整的蛋白质可以通过凝胶电泳分离。十二烷基硫酸钠聚丙烯酰胺凝胶电泳(SDS-PAGE)结合激光烧蚀ICP-MS对¹²⁷I的检测表明,在分子量为110 kDa、40 kDa、27 kDa、20 kDa和10 kDa的蛋白条带中存在碘。二维等电聚焦-十二烷基硫酸钠聚丙烯酰胺凝胶电泳(2D IEF-SDS PAGE)结合激光烧蚀ICP-MS对¹²⁷I的成像,能够检测到碱性pH值范围内的5个含碘蛋白质斑点。