Studies on the denaturation of muscle proteins under high pressure.

高压下肌肉蛋白变性的研究。

• 批准号: 10460118
• 负责人: SUZUKI Atsushi
• 金额: $ 8.77万
• 依托单位: NIIGATA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B).
• 财政年份: 1998
• 资助国家: 日本
• 起止时间: 1998 至 2000
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-10460118/
• 关键词: high pressure treatment; denaturation of actin; denaturation of myosin; mitochondria; intramuscular connective tissue; proteasome; high pressure-thawing; β-Lactoglobulin; 超高圧; 筋肉タンパク質; ミオシンの圧力変性; トロポミオシンの圧力変性; 質量中心; α-ヘリックス; ミオシンATPアーゼ; 加圧解

The denaturation of muscle proteins under high pressure was investigated by histological, biochemical and physicochemical methods.Actin1) The rate of dissociation of nucleotides from actin molecule, disappearance of the characterized ^1H NMR signal at 2.055 ppm and loss in biochemical activities (DNAasel inhibition capacity, polymerizability) at 300 MPa were almost identical. suggesting that the rapid collapse of steric structure around the upper region called "pointed end" of actin molecule following the dissociation of the bound nucleotide.2) The decreases of α-helix content were observed with the pressure applied and the denaturation was irreversible in the actin pressurized at 300 MPa or more.Myosin and Tropomyosin1) The structural changes in myosin molecule pressurized at 200 MPa or more were irreversible.2) Tropomyosin which is resistive to acid and heat was also resistive to high pressure.Mitochondria and Connective Tissue1) The decrease of Ca^<2+> uptaking ability seemed to be correlated with the decrease of ATPase activity which may be caused by the release of ATPase subunits (α β) from mitochondria by pressure.2) The deformation of the honeycomb-like structure of endomysium was accelerated with increase of pressure applied, but the changes in the extractability of PGs were not observed in the pressurized muscle.ProteasomeThe high pressure treatments up to 50〜100MPa caused the activation of the proteasome. This activation is probably due to the increase of the interaction of the substrate and active site of the proteasome unfolded by high pressure treatment.β-Lactoglobulin (BLg) and High Pressure-Thawing1) No difference in the ^1NMR spectrum was observed between pressurized and unpressurized BLg. NMR detection of the H/D exchange of aromatic residues indicated that the conformation at the vicinity of tryptophan residues can be refolded after the release of pressure.2) It was cleared that the high pressure-thawing was applicable to meat processing.

通过组织学、生物化学和物理化学方法研究了高压下肌肉蛋白的变性。肌动蛋白1) 肌动蛋白分子中核苷酸的解离速率、2.055 ppm 处特征^1H NMR 信号的消失以及生化活性的损失（DNAasel 抑制能力） ，聚合性）在 300 MPa 下几乎相同的塌陷，表明称为“尖端”的上部区域周围的空间结构快速。结合的核苷酸解离后肌动蛋白分子。2）随着施加压力，观察到α-螺旋含量减少，并且在300 MPa或以上加压的肌动蛋白中，变性是不可逆的。肌球蛋白和原肌球蛋白1）肌球蛋白分子的结构变化2)原肌球蛋白耐酸耐热，耐高温，200MPa以上加压是不可逆的。线粒体与结缔组织1) Ca^<2+>摄取能力的降低似乎与ATPase活性的降低有关，这可能是由于压力从线粒体释放ATPase亚基(α β)引起的。2)随着施加压力的增加，肌内膜蜂窝状结构的变形加速，但在加压肌肉中未观察到 PG 提取率的变化。蛋白酶体高压处理至50～100MPa引起蛋白酶体的活化，这种活化可能是由于高压处理使蛋白酶体展开的底物和活性位点的相互作用增加。β-乳球蛋白(BLg)和高压解冻1)无差异。在^1NMR谱中观察到加压和非加压BLg之间的芳香族残基的H/D交换的NMR检测表明附近的构象。释放压力后色氨酸残留物可重新折叠。2)明确高压解冻适用于肉类加工。

项目成果

Y.Ikeuchi: "High pressure-induced textural changes of food proteins"Nippon Nougei Kagakukaishi. Vol.74(5). 612-615 (2000)

Y.Ikeuchi：“高压引起的食物蛋白质的结构变化”Nippon Nougei Kagakukaishi。

Y.Ueno, Y.Ikeuchi, and A.Suzuki: "Effects of high pressure treatments on intramuscular connective tissue"Meat Sci.. Vol.52. 143-150 (1999)

Y.Ueno、Y.Ikeuchi 和 A.Suzuki：“高压治疗对肌内结缔组织的影响”Meat Sci.. Vol.52。

Shuhei Yamamoto: "Trends in High Pressure Bioscience and Technology"Elsevier Science B.V.. 6 (2001)

Shuhei Yamamoto：“高压生物科学与技术的趋势”Elsevier Science B.V.. 6 (2001)

池内義秀: "高圧による食品タンパク質の物性変化"日本農芸化学会誌. 74巻・5号. 612-615 (2000)

Yoshihide Ikeuchi：“高压引起的食品蛋白质物理特性的变化”，日本农业化学学会杂志，第 74 卷，第 5 期。612-615（2000 年）。

Atsushi Suzuki: "Pressure-induced changes in the connectin/titin localization in the myofibrils revealed by immunoelectron microscopy"Meat Science. (印刷中). (2001)

Atsushi Suzuki：“通过免疫电子显微镜揭示压力引起的肌原纤维中连接蛋白/肌联蛋白定位的变化”《肉类科学》（出版中）。