异硫氰酸烯丙酯对产气荚膜梭菌的抑菌作用及在熟猪肉糜中的应用

黎琳莹; 林以琳; 温耀升; 廖彩虎; 余以刚

doi:10.13386/j.issn1002-0306.2023020200

异硫氰酸烯丙酯对产气荚膜梭菌的抑菌作用及在熟猪肉糜中的应用

Inhibitory Effect of Allyl Isothiocyanate on Clostridium perfringens and Its Application of Cooked Pork

摘要

摘要: 本文旨在研究异硫氰酸烯丙酯（Allyl isothiocyanate，AITC）对产气荚膜梭菌（Clostridium perfringens，C. perfringens）的抑菌效果及作用机制。首先，通过测定最小抑菌浓度（Minimum inhibition concentration，MIC）、绘制生长曲线评估AITC对C. perfringens的抑菌效果，并采用扫描电镜观察细胞形态、碘化丙啶染色实验测定细胞膜完整性，评估AITC对C. perfringens细胞膜的影响；进一步通过SDS-PAGE图谱分析和ATP酶活力测定研究AITC对C. perfringens细胞代谢的影响；最后，研究了AITC对熟猪肉糜中C. perfringens的抑制效果。结果表明，AITC可有效抑制C. perfringens的生长，其MIC为0.1 μL/mL；AITC引起C. perfringens细胞膜破裂、凹陷等形变，导致细胞膜完整性丧失，且膜损伤的程度随AITC浓度的升高而增强；AITC显著（P<0.05）降低了C. perfringens的蛋白质含量和ATP酶活性，进而影响细胞的正常代谢；此外，添加0.1%~0.4%的AITC能够显著（P<0.05）抑制熟猪肉糜中C. perfringens的生长。综上，AITC可以通过破坏C. perfringens的细胞膜和干扰蛋白质代谢达到抑菌效果，本研究为天然抑菌剂在肉类行业中的应用提供了一定的理论基础。

Abstract: The study aimed to investigate the inhibitory activity and mechanism of action of allyl isothiocyanate (AITC) against Clostridium perfringens (C. perfringens). Firstly, the inhibitory effect of AITC on C. perfringens was evaluated by determining the minimum inhibitory concentration (MIC) and plotting the growth curve. Secondly, the effect of AITC on C. perfringens cell membranes was assessed by scanning electron microscopy to observe cell morphology and by measuring cell membrane integrity with a propidium iodide staining assay. In addition, the impact of AITC on the metabolism of C. perfringens was investigated using SDS-PAGE profile and an ATPase activity assay. Finally, the inhibitory effect of AITC on C. perfringens in cooked ground pork was examined. The experimental results showed that AITC could effectively inhibit the growth of C. perfringens, and the MIC of AITC was determined to be 0.1 μL/mL. AITC was able to induce cell membrane deformations, such as rupture and depression, resulting in the loss of C. perfringens cell membrane integrity, and the degree of membrane damage increased with AITC concentration. Meanwhile, AITC could reduce the protein content and ATPase activity of C. perfringens, which had an impact on normal cellular metabolism. Furthermore, the addition of 0.1%~0.4% AITC significantly inhibited the growth of C. perfringens in cooked ground pork (P<0.05). In conclusion, AITC could achieve bacterial inhibition by disrupting the cell membrane of C. perfringens and interfering with protein metabolism, and this study offered a theoretical foundation for the use of natural bacterial inhibitors in the meat industry.