食品科学 ›› 2019, Vol. 40 ›› Issue (6): 1-8.doi: 10.7506/spkx1002-6630-20180326-353

• 食品化学 •    下一篇

翡翠贻贝副肌球蛋白的特性及在模拟胃肠液中的消化

邹 睿1,张凌晶1,2,钟 婵1,翁 凌1,2,林丽云1,李钰金3,刘光明1,2,曹敏杰1,2,*   

  1. 1.集美大学食品与生物工程学院,福建 厦门 361021;2.水产品深加工技术国家地方联合工程研究中心,福建 厦门 361021;3.泰祥集团 山东省海洋食品营养研究院,山东 荣成 264309
  • 出版日期:2019-03-25 发布日期:2019-04-02
  • 基金资助:
    国家自然科学基金面上项目(31471640);福建省科技计划项目(2017N5011)

Characterization and Simulated Gastrointestinal Digestion of Paramyosin from Perna viridis

ZOU Rui1, ZHANG Lingjing1,2, ZHONG Chan1, WENG Ling1,2, LIN Liyun1, LI Yujin3, LIU Guangming1,2, CAO Minjie1,2,*   

  1. 1. College of Food and Biological Engineering, Jimei University, Xiamen 361021, China; 2. National & Local Joint Engineering Research Center of Processing Technology for Aquatic Products, Xiamen 361021, China; 3. Shandong Marine Food Nutrition Research Institute, Taixiang Group Company, Rongcheng 264309, China
  • Online:2019-03-25 Published:2019-04-02

摘要: 为探究贝类副肌球蛋白(paramyosin,PM)的热稳定性、pH值稳定性及其在模拟胃肠液中的消化特性,以翡翠贻贝(Perna viridis)为对象,利用硫酸铵盐析和羟基磷灰石柱层析等方法,从肌肉中纯化PM,采用肽质量指纹图谱法对其进行鉴定,利用圆二色谱测定其二级结构及热变性温度。结果显示,翡翠贻贝PM分子质量为99.5 kDa;肽质量指纹图谱分析获得26 个肽段,共330 个氨基酸残基,与地中海贻贝(Mytilus galloprovincialis)PM的序列一致性达到100%,表明纯化的蛋白为PM。圆二色谱结果显示,PM呈现典型的α-螺旋结构,其热变性温度为(56.3±0.2)℃。在30~100 ℃热处理30 min,PM未出现沉淀聚集现象,在pH 6~11范围内也较稳定,但当pH≤5时稳定性差,出现沉淀聚集现象。与胃蛋白酶、胰蛋白酶及胰凝乳蛋白酶对PM的单独消化作用相比,3 种酶连续作用可使PM有效降解,但仍有分子质量30 kDa的片段未被完全消化。本研究表明,翡翠贻贝PM具有较好的耐热性及耐消化性,为贻贝深加工及PM的后续研究提供一定理论参考。

关键词: 副肌球蛋白, 翡翠贻贝, 分离纯化, 稳定性, 模拟胃肠液消化

Abstract: In order to investigate the thermal stability, pH stability and digestion characteristics of paramyosin (PM) in simulated gastrointestinal fluids, we purified PM to homogeneity from the muscle of Perna viridis by consecutive ammonium sulfate fractionation and hydroxyapatite chromatography, and we further identified it by peptide mass fingerprinting (PMF). Circular dichroism (CD) spectroscopy was employed to measure its secondary structure and thermal denaturation temperature. SDS-PAGE was carried out to explore its thermal stability, pH stability and digestion characteristics in simulated gastrointestinal fluids. PM showed a single band corresponding to 99.5 kDa on SDS-PAGE. Using peptide mass fingerprinting, a total of 26 peptide fragments, including 330 amino acid residues, were obtained from purified PM, which revealed 100% identity to PM from Mytilus galloprovincialis, indicating that the purified protein is PM. CD spectral analysis demonstrated that PM had a typical α-helix structure with thermal denaturation temperature (Td) of (56.3 ± 0.2) ℃. No obvious precipitate was observed when PM was heated in the temperature range from 30 to 100 ℃ for 30 min. PM was stable between pH 6.0–11.0. However, it was unstable below pH 5.0 and aggregated. PM could only be partially hydrolyzed by pepsin, trypsin or chymotrypsin individually, while it was effectively degraded by continuous hydrolysis with the three proteinases. However, protein bands with molecular mass of approximately 30 kDa remained undigested. In conclusion, PM from Perna viridis is thermally stable and resistant to gastrointestinal digestion. This work provides a valuable theoretical basis for mussel processing and further study on PM.

Key words: paramyosin, Perna viridis, purification, stability, simulated gastrointestinal digestion

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