Zhang Kunfeng, Huang Yuzhe, Wu Qianzhen, Guo Wenhua, Chen Hao, Zhang Wenna, Li Yong, Lu Yongming, Wu Qingxi, Pan Wenjuan, Chen Lei, Chen Yan
School of Life Sciences, Anhui University, Hefei, Anhui, China.
School of Life Sciences, Anhui University, Hefei, Anhui, China; Key Laboratory of Ecological Engineering and Biotechnology of Anhui Province and Anhui Key Laboratory of Modern Biomanufacturing, Hefei, Anhui, China.
Int J Biol Macromol. 2022 May 15;207:750-759. doi: 10.1016/j.ijbiomac.2022.03.086. Epub 2022 Mar 19.
The objectives of this study were (1) to prepare Armillariella tabescens mycelia polysaccharides (PAT) with remarkably growth inhibitory effect on typical food-borne pathogenic bacteria using a green and efficient polyamide method and (2) to explore the antibacterial mechanism of PAT for use as a natural antibacterial agent. The sugar and uronic acid contents of PAT were 93.41% and 12.24%, respectively. PAT could inhibit the growth of Escherichia coli, Proteus vulgaris, Bacillus subtilis, and Staphylococcus aureus cells, with minimum inhibitory concentrations of 0.5, 1.0, 4.0, and 4.0 mg/mL, respectively. Ultra-high-resolution field emission scanning electron microscopy and high-resolution transmission electron microscopy analysis revealed cell wall and membrane rupture of E. coli treated with PAT. Further, 0.5-4.0 mg/mL PAT was found to significantly (P < 0.01) and concentration-dependently increase the conductivity of the broth, exudation of the intracellular protein, and alkaline phosphatase and β-galactosidase activities. Confocal laser scanning microscopy revealed morphological changes in E. coli DNA after PAT treatment and intracellular reactive oxygen species accumulation; flow cytometry revealed E. coli cell apoptosis. Our findings provide a theoretical basis and technical support for the development of PAT as a natural antibacterial product.
(1)使用绿色高效的聚酰胺法制备对典型食源致病菌具有显著生长抑制作用的亮菌菌丝体多糖(PAT);(2)探究PAT作为天然抗菌剂的抗菌机制。PAT的糖含量和糖醛酸含量分别为93.41%和12.24%。PAT能够抑制大肠杆菌、普通变形杆菌、枯草芽孢杆菌和金黄色葡萄球菌细胞的生长,其最低抑菌浓度分别为0.5、1.0、4.0和4.0mg/mL。超高分辨率场发射扫描电子显微镜和高分辨率透射电子显微镜分析显示,经PAT处理的大肠杆菌细胞壁和细胞膜破裂。此外,发现0.5 - 4.0mg/mL的PAT能显著(P < 0.01)且呈浓度依赖性地增加肉汤的电导率、细胞内蛋白质的渗出以及碱性磷酸酶和β-半乳糖苷酶的活性。共聚焦激光扫描显微镜显示PAT处理后大肠杆菌DNA的形态变化以及细胞内活性氧的积累;流式细胞术显示大肠杆菌细胞凋亡。我们的研究结果为开发PAT作为天然抗菌产品提供了理论依据和技术支持。