HPTS结合蛋清溶菌酶对枯草杆菌芽孢的灭活作用
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(宁夏大学食品科学与工程学院 银川 750021)

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国家自然科学基金项目(32260633,31760474)


Inactivation of High-pressure Thermal Sterilization Combining with Egg White Lysozyme on Bacillus subtilis Spores
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(College of Food Science and Engineering, Ningxia University, Yinchuan 750021)

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    摘要:

    本文研究了高压热杀菌技术(HPTS)结合蛋清溶菌酶处理对枯草杆菌芽孢的杀灭作用。采用 HPTS(200 MPa-25,65,75 ℃)结合蛋清溶菌酶(0.05%,0.1%,0.3%)保压处理20 min,处理后测定枯草杆菌芽孢的失活量、蛋白质与核酸的泄漏量、电导率、细胞膜Na+/K+-ATPase活性变化、芽孢的形态结构变化、蛋白质二级结构稳定性变化、内膜通透性变化。结果表明,HPTS结合溶菌酶对芽孢具有明显的协同杀灭作用,200 MPa/75 ℃结合0.3%溶菌酶处理后,芽孢存活浓度下降3.91 lg (CFU/mL)。HPTS结合溶菌酶处理后芽孢蛋白质、核酸泄漏量及上清液电导率都显著增加,Na+/K+-ATPase活性显著降低,芽孢蛋白质稳定性下降,芽孢表面显示出极度的粗糙,伴随着严重的塌陷皲裂,结构发生了明显变化。流式细胞术分析发现,芽孢内膜的通透性显著增加。结论:HPTS结合蛋清溶菌酶处理抑制了芽孢代谢途径中关键酶的活性,同时破坏了枯草杆菌芽孢的芽孢壁、内膜等结构,改变了芽孢的生理特性,最终导致芽孢失活。

    Abstract:

    This study aimed to investigate the inactivation effect of HPTS combining with egg white lysozyme on the spores of Bacillus subtilis. The spores were treated with pressure 200 MPa at 25, 65 ℃ and 75 ℃, and egg white lysozyme at 0.05%, 0.1% and 0.3%. After treatment, the inactivation rate of Bacillus subtilis spores, leakage of nucleic acid and protein, the change in the cell membrane ATPase activity, the changes in inner membrane permeability of spores and the damage on protein of the spores were determined. The results showed that HPTS and egg white lysozyme synergistically inactivated the spores, and the survival concentration of spores decreased by 3.91 lg (CFU/mL) after 200 MPa/75 ℃ combining with 0.3% lysozyme treatment. HPTS combining with lysozyme treatment significantly increased the protein and nucleic acid leakage and conductivity of the spores, significantly decreased the Na+/K+-ATPase activity, decreased the stability of the budding spore proteins, and significant increases in the permeability of the inner membrane of the spores. In summary, HPTS combining with egg white lysozyme treatment inhibited the activity of key enzymes in the spore metabolic pathway, while disrupting the spore wall, inner membrane and other structures of Bacillus subtilis budding spores, changed the physiological properties of spores, and eventually led to spore inactivation.

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辛伟山,刘月,李佳佳,武思睿,王旭娟,王传发,章中. HPTS结合蛋清溶菌酶对枯草杆菌芽孢的灭活作用[J].中国食品学报,2024,24(3):112-121

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  • 收稿日期:2023-03-24
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  • 在线发布日期: 2024-04-16
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