目前,关于乳酸菌抗真菌肽抑制真菌的作用机制研究相对较少。该试验以副干酪乳杆菌ALAC-4产生的抗真菌肽APT为研究对象,采用活细胞计数法评价了其最小抑菌浓度为15.63 mg/mL,抑菌率为65%。以粘红酵母菌作为指示菌,对抗菌肽的抑菌机理进行了初步研究。测定了抗菌肽处理后粘红酵母的β-1,3-葡聚糖和几丁质的暴露量,结果表明APT可能破坏了粘红酵母细胞壁的结构;K+泄漏实验结果显示,粘红酵母菌胞内K＋释放量随APT浓度的升高显著增加;扫描电镜和透射电镜显示APT可使菌体细胞表面出现皱缩、孔洞,细胞质出现空泡现象;通过碘化丙啶和5(6)-羧基二乙酸荧光素琥珀酰亚胺酯标记实验结果表明APT处理粘红酵母细胞内酶失活状况与细胞膜的破损并非同步进行。综上可知抗菌肽APT对粘红酵母细胞壁膜以及胞内酶系造成影响从而起到抑菌作用。
At present, there is relatively little information on the inhibiting fungi mechanism of antifungal peptide produced by lactic acid bacteria.Using the antibacterial peptide APT produced by Lactobacillus paracasei ALAC-4 as research object in the research, the minimum inhibitory concentration and the antibacterial rate were confirmed to be 15.63 mg/mL and 65% respectively by the viable cell counting method.The antifungal mechanism of APT was studied using Rhodotorula glutinis as an indicator strain.By measuring the exposure of β-1,3-glucan and chitin in R.glutinis after treatment with the peptide.Results showed that APT could destroy the structure of the cell wall of R.glutinis.The results of the K+ leakage experiment showed that the amount of intracellular K+ release increased significantly with the increase of APT concentration. Scanning electron microscope and transmission electron microscope showed that APT could cause shrinkage and holes on surface of yeast cell, moreover cause vacuoles in the cytoplasm.It was found by propidium Iodide and carboxyfluorescein diacetate labeling experiments that the intracellular enzyme inactivation and cell membrane damage of R.glutinis treated with APT were not synchronized.In summary, it can be seen that APT affect the cell wall membrane and intracellular enzymes of R.glutinis, thereby exert inhibiting effects.
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