食品科学 ›› 2019, Vol. 40 ›› Issue (21): 107-114.doi: 10.7506/spkx1002-6630-20190304-036

• 食品工程 • 上一篇    下一篇

超高压结合胰蛋白酶消减虾原肌球蛋白致敏性及其抗原线性表位残留

胡志和,王丽娟,薛璐,刘平,贾莹,鲁丁强   

  1. (天津商业大学生物技术与食品科学学院,天津市食品生物技术重点实验室,天津 300134)
  • 出版日期:2019-11-15 发布日期:2019-12-02
  • 基金资助:
    国家自然科学基金面上项目(31271841);天津市高等学校创新团队项目(TD13-5087)

Allergenicity Reduction of Shrimp Tropomyosin by High Hydrostatic Pressure Treatment Combined with Enzymatic Hydrolysis and Its Linear Epitope Residues

HU Zhihe, WANG Lijuan, XUE Lu, LIU Ping, JIA Ying, LU Dingqiang   

  1. (Tianjin Key Laboratory of Food Biotechnology, School of Biotechnology and Food Science, Tianjin University of Commerce, Tianjin 300134, China)
  • Online:2019-11-15 Published:2019-12-02

摘要: 比较常压下酶水解与超高压下酶水解虾原肌球蛋白(tropomyosin,TM)的致敏性差异及线性表位的残留,探讨超高压下酶水解TM增强致敏性消减效果的原因。采用间接酶联免疫吸附测定法检测TM致敏性,傅里叶变换红外光谱和荧光光谱法检测TM二级和三级结构;以致敏性(OD492 nm)为指标,确定常压和超高压下酶水解TM的条件;采用质谱法检测水解片段的氨基酸序列。结果表明,在40 ℃、200 MPa下,保压时间30 min有利于胰蛋白酶活力的提高;压力在100~600 MPa内,TM致敏性随压力升高而降低,其致敏性与二级结构中β-转角的相对含量及三级结构的变化有关;在常压下(40 ℃、酶添加量3 000 U/g、水解30 min),TM水解产物的致敏性消减率为89%,水解产物中含8~16 个氨基酸残基的片段数量占76.8%,且线性表位的消减率为60.0%~66.7%;在超高压下(40 ℃、酶添加量3 000 U/g、200 MPa下水解30 min),TM水解产物的致敏性消减率为98%,水解产物中含8~16 个氨基酸残基的片段数量占93.3%,且线性表位的消减率为88.9%~90.0%。因此,超高压可以促进酶水解TM,有利于线性表位的消除,从而降低水解产物的致敏性。

关键词: 原肌球蛋白, 超高静压, 胰蛋白酶, 致敏性, 线性表位

Abstract: The objective of this work was to compare the differences in the allergenicity and linear epitope residues of trypsin hydrolysates of tropomyosin (TM) prepared under normal and high pressure conditions and to discuss the reason why allergenicity reduction of hydrolyzed TM is enhanced under high pressure. The allergenicity was detected by indirect enzyme-linked immunosorbent assay (iELISA), and the secondary and tertiary structures were investigated by infrared and fluorescence spectroscopies. Optimal hydrolysis conditions under normal and high pressure were determined based on the binding capacity of hydrolysate to antibody (OD492 nm). Mass spectrometry (MS) was used for detecting the amino acid sequences of hydrolytic fragments. The results showed that trypsin activity was enhanced after treatment for 30 min under 200 MPa at 40 ℃. The allergenicity of high-pressure treated TM decreased with increasing pressure from 100 to 600 MPa, and it was related to changes in the relative content of β-turn structure and in tertiary structure. The allergenicity of TM was reduced by 89% and 98% after 30 min hydrolysis with 3 000 U/g of trypsin at 40 ℃ under normal pressure and 200 MPa, respectively. The percentage of fragments containing 8–16 amino acid residues in the hydrolysates was 76.8% and 93.3%, respectively, and the linear epitopes were reduced by 60.0%–66.7% and 88.9%–90.0%, respectively. Therefore, high pressure treatment could promote the enzymatic hydrolysis of TM, facilitating the elimination of linear epitopes and reducing the allergenicity of hydrolyzed products.

Key words: tropomyosin, high hydrostatic pressure, trypsin, allergenicity, linear epitope

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