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植物乳杆菌胞外多糖纳米银的结构、物理性质及抑菌活性。

Structures, physical properties and antibacterial activity of silver nanoparticles of Lactiplantibacillus plantarum exopolysaccharide.

机构信息

Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, China.

Engineering Research Center of Agricultural Microbiology Technology, Ministry of Education & Heilongjiang Provincial Key Laboratory of Plant Genetic Engineering and Biological Fermentation Engineering for Cold Region & Key Laboratory of Microbiology, College of Heilongjiang Province & School of Life Sciences, Heilongjiang University, Harbin 150080, China; Hebei University of Environmental Engineering, Hebei Key Laboratory of Agroecological Safety, Qinhuangdao 066102, China.

出版信息

Int J Biol Macromol. 2024 Apr;263(Pt 2):130083. doi: 10.1016/j.ijbiomac.2024.130083. Epub 2024 Feb 29.

Abstract

Lactic acid bacteria (LAB) exopolysaccharide (EPS) has good water absorption, high viscosity, good stability, so it was widely used in probiotics fields. In this study, EPS-producing LAB strain Lactiplantibacillus plantarum HDL-03 was isolated and identified. Moreover, the HDL-03 EPS was used as a stabilizer and mixed with AgNO to synthesize a novel nanoparticle AgNPs whose structure and properties were explored. The monosaccharide composition and molecular weight indicated that HDL-03 EPS was a heteropolysaccharide composed of mannose and glucose. Fourier transform infrared spectroscopy (FT-IR), nuclear magnetic resonance (NMR) spectroscopy analysis and methylation results jointly proved it was a heteropolysaccharide containing 1,3-Manp and 1,6-Glcp. The X-Ray diffraction (XRD) results showed that this EPS has an amorphous structure, while the synthesized AgNPs have crystalline properties. The scanning electron microscopy (SEM) and transmission electron microscopy (TEM) results indicated EPS had a smooth and dense sheet structure, while the surface of AgNPs became rougher and large holes appeared after synthesis. Zeta particle size analysis suggested that the particle size of AgNPs increased by 36.63 nm compared to HDL-03 EPS. FT-IR analysis exhibited that the position of the characteristic peaks of AgNPs changed. The OH moving from a wavelength of 3388.49 cm to a wavelength of 3316.79 cm and telescopic vibration peak changed from 1356.07 cm to 1344.22 cm. A plate inhibition test revealed the effect of different concentrations of EPS and AgNO synthesized AgNPs on the diameter of inhibition circle produced by the indicator bacteria Escherichia coli and Staphylococcus aureus. Furthermore, AgNPs were applied to the indicator bacteria, which the minimum inhibitory concentration (MIC), time-inhibitory curve, and changes in extracellular conductivity, nucleic acids, proteins, ATP, and lactate dehydrogenase (LDH) levels were determined. The AgNPs inhibited the growth of E. coli and S. aureus and exhibited outstanding antimicrobial properties. With the increase of treatment time, the degree of cell membrane damage increased, the permeability enhanced, and the intracellular substances leaked. These results indicate that HDL-03 EPS has good potential for applications in the production of food packaging, antimicrobials, catheters, textiles and coatings.

摘要

乳酸菌(LAB)胞外多糖(EPS)具有良好的吸水性、高粘度、良好的稳定性,因此被广泛应用于益生菌领域。本研究从植物乳杆菌中分离鉴定出一株产 EPS 的菌株 Lactiplantibacillus plantarum HDL-03,并将其 EPS 作为稳定剂与 AgNO 混合合成新型纳米银颗粒 AgNPs,对其结构和性能进行了探讨。单糖组成和分子量分析表明,HDL-03 EPS 是由甘露糖和葡萄糖组成的杂多糖。傅里叶变换红外光谱(FT-IR)、核磁共振(NMR)波谱分析和甲基化结果共同证明,这是一种含有 1,3-Manp 和 1,6-Glcp 的杂多糖。X 射线衍射(XRD)结果表明,该 EPS 具有无定形结构,而合成的 AgNPs 具有结晶性。扫描电子显微镜(SEM)和透射电子显微镜(TEM)结果表明,EPS 具有光滑致密的片状结构,而 AgNPs 表面在合成后变得更加粗糙,出现大孔。Zeta 粒径分析表明,与 HDL-03 EPS 相比,AgNPs 的粒径增加了 36.63nm。FT-IR 分析表明,AgNPs 的特征峰位置发生了变化。OH 从 3388.49cm 处的移动到 3316.79cm 处,伸缩振动峰从 1356.07cm 处变为 1344.22cm 处。平板抑菌试验表明,不同浓度 EPS 和 AgNO 合成的 AgNPs 对指示菌大肠杆菌和金黄色葡萄球菌产生的抑菌圈直径的影响。此外,将 AgNPs 应用于指示菌,测定其最小抑菌浓度(MIC)、时间抑制曲线以及细胞外电导率、核酸、蛋白质、ATP 和乳酸脱氢酶(LDH)水平的变化。AgNPs 抑制了大肠杆菌和金黄色葡萄球菌的生长,表现出优异的抗菌性能。随着处理时间的增加,细胞膜损伤程度增加,通透性增强,细胞内物质泄漏。这些结果表明,HDL-03 EPS 在食品包装、抗菌剂、导管、纺织品和涂料的生产中具有良好的应用潜力。

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