食品科学 ›› 2023, Vol. 44 ›› Issue (5): 29-37.doi: 10.7506/spkx1002-6630-20220226-231

• 基础研究 • 上一篇    

抗菌肽zp37抑制果汁中单核细胞增生李斯特菌的活性及其作用机制

任俊和,曾平,陈思睿,易兰花   

  1. (1.西南大学食品科学学院,重庆 400715;2.香港中文大学药剂学院,香港 999077;3. 重庆市开州区疾病预防控制中心,重庆 405499)
  • 发布日期:2023-03-23
  • 基金资助:
    国家自然科学基金青年科学基金项目(32102032)

Inhibitory Effect of Antimicrobial Peptide zp37 on Listeria monocytogenes in Fruit Juice and Its Action Mechanism

REN Junhe, ZENG Ping, CHEN Sirui, YI Lanhua   

  1. (1. College of Food Science, Southwest University, Chongqing 400715, China;2. School of Pharmacy, The Chinese University of Hong Kong, HongKong 999077, China;3. Chongqing Kaizhou Center for Disease Control and Prevention, Chongqing 405499, China)
  • Published:2023-03-23

摘要: 单核细胞增生李斯特菌(简称单增李斯特菌)作为一种常见的食源性致病菌,是影响果汁食用安全性的主要威胁因子。本实验设计并获得一种抗菌肽zp37,通过测定抗菌肽zp37最小抑菌浓度(minimum inhibitory concentration,MIC),分析抗菌肽zp37处理后单增李斯特菌的生长曲线、膜电位、膜完整性、细胞微观结构、细胞聚集率、DNA结合情况以及胞内活性氧等,研究抗菌肽zp37抑制果汁中单增李斯特菌的活性及其作用机制。结果表明,抗菌肽zp37对单增李斯特菌的MIC为16 μg/mL,当其质量浓度大于4 MIC时,单增李斯特菌生长受到明显抑制。同时,8 MIC的抗菌肽zp37处理1 h能杀死绝大部分的单增李斯特菌,且在7 d贮藏期内,2 MIC的抗菌肽zp37能将草莓汁、猕猴桃汁和苹果汁中的单增李斯特菌控制到检测限以下。抗菌肽zp37处理后,单增李斯特菌细胞膜离子通透性增加、膜电位消失,细胞膜完整性被破坏,细胞出现明显的凹陷变形,细胞间发生聚集。荧光标记示踪法结果显示,抗菌肽zp37同时作用于单增李斯特菌的细胞膜和细胞质。在细胞质中,当抗菌肽zp37质量浓度大于DNA质量浓度的1/4时,两者会发生明显结合甚至导致DNA沉淀。此外,抗菌肽zp37能诱导单增李斯特菌产生活性氧,且与质量浓度成正比。综上,抗菌肽zp37对果汁中的单增李斯特菌抑制效果明显,本研究可为抗菌肽作为生物防腐剂应用于果汁提供一定的科学依据。

关键词: 抗菌肽;果汁;单核细胞增生李斯特菌;抑菌活性;作用机制

Abstract: As a common foodborne pathogen, Listeria monocytogenes is a major threat to the safety of fruit juice. In this study, The activity and action mechanism of antimicrobial peptide zp37 in inhibiting L. monocytogenes in fruit juice were studied by measuring its minimum inhibitory concentration (MIC), and analyzing its growth curve, membrane potential, membrane integrity, cell microstructure, cell aggregation rate, DNA binding, intracellular reactive oxygen species (ROS) level after zp37 treatment. The results showed that the MIC value of antimicrobial peptide zp37 against L. monocytogenes was 16 μg/mL, and the growth of L. monocytogenes was significantly inhibited by zp37 at concentrations greater than 4 MIC. After treatment with the peptide at 8 MIC concentration for 1 h, L. monocytogenes was almost completely killed. The peptide at 2 MIC concentration kept the number of L. monocytogenes in strawberry, kiwifruit and apple juice below the detection limit for seven days. After treatment with zp37, the cell membrane permeability of L. monocytogeneswas increased, the membrane potential was lost, the cell membrane integrity was destroyed, the cell surface became obviously sunken and deformed, and cell aggregation occurred. The results of fluorescent labeling showed that zp37 could simultaneously act on the cell membrane and cytoplasm of L. monocytogenes. In the cytoplasm, when zp37 concentration was greater than one-fourth of the DNA concentration, they could bind significantly and even DNA was precipitated. In addition, zp37 induced L. monocytogenes to produce ROS in a positive dose-dependent manner. The above results showed that antimicrobial peptide zp37 had anobvious inhibitory effect on L. monocytogenes in fruit juice, which can provide a scientific basis for the application of antimicrobial peptides as biological preservatives in fruit juice.

Key words: antimicrobial peptide; fruit juice; Listeria monocytogenes; antibacterial activity; action mechanism

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