Abstract：
Parvalbumin (PV) is a major allergen in fish, and traditional treatments cannot reduce its sensitization. The effects of dense-phase carbon dioxide (DPCD) treatment on the sensitization and spatial structure of PV in Trachinotus ovatus were evaluated in this study. Western blotting and indirect ELISA were used to determine the allergenicity changes and spatial conformations of PV treated by DPCD. Tris-tricine-SDS-PAGE, circular dichroism, surface hydrophobicity, endogenous fluorescence, UV spectrophotometry, free amino group, total sulfhydryl group and SEM analyses were applied to characterize PV structure. The results showed that DPCD treatment (15 MPa, 30 min, 50 °C) could reduce PV-induced allergic reactions by 39-41 %, which destroyed the normal conformational epitopes and reduced the risk of PV-induced allergy. The secondary structure changed from ordered to disordered with a decreased content of α-helical groups, while the internal hydrophobic groups were exposed. The total sulfhydryl group content decreased significantly (P < 0.05). The surface hydrophobicity and ultraviolet absorption spectrum were enhanced, and the endogenous fluorescence peak shifted to a long wavelength. Meanwhile, the content of free amino groups increased significantly (P < 0.05). This study could provide a theoretical basis and a promising technical approach for reduction of PV allergenicities.
摘要：
小白蛋白（PV）是鱼类的主要过敏原，传统的治疗方法不能降低其敏感性。在这项研究中，评估了密相二氧化碳（DPCD）处理对TrachinotusovatusPV的敏化和空间结构的影响。采用Western印迹和间接ELISA检测DPCD处理PV的变应原性变化和空间构象。Tris-tricine-SDS-PAGE,圆二色性,表面疏水性，内源性荧光，紫外分光光度法,游离氨基,总巯基和SEM分析用于表征PV结构。结果表明,DPCD处理(15MPa,30分钟,50°C）可将PV引起的过敏反应减少39-41％，这破坏了正常的构象表位，降低了PV诱导的过敏风险。随着α-螺旋基团含量的减少，二级结构从有序变为无序，而内部疏水基团被暴露。总巯基含量明显下降(P<0.05)。表面疏水性和紫外吸收光谱增强,和内源性荧光峰移动到一个长波长。同时,游离氨基含量显著增加(P<0.05)。本研究可为降低肺静脉变应原性提供理论依据和技术途径。
