Optimization of the Conditions for the Transformation of a Bacillus subtilis Strain L11 to Prepare Nano Selenium and Its Preliminary Application in Sheep Feed

Wenxin Guo, Xinyu Shi, Lu Wang, Xin Cong, Shuiyuan Cheng, Linling Li, Hua Cheng
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Abstract

Selenium nanoparticles (SeNPs) have greater bioavailability and safety than inorganic selenium, and was widely used in medical, agricultural, nutritional supplements, and antibacterial fields. The present study screened a strain L11 producing SeNPs from a selenium rich dairy cow breeding base in Hubei Province, China. The strain was identified as Bacillus subtilis through physiological, biochemical, and molecular biology analysis. By adjusting the cultivation conditions, the experiment determined the ideal parameters for L11 to efficiently produce SeNPs. These parameters include a pH value of 6, a cultivation temperature of 37 °C, a concentration of 4 mmol/L Na2SeO3, and a cultivation of 48 h. X-ray Photoelectron Spectroscopy (XPS), Scanning Electron Microscope-Energy Dispersive Spectroscopy (SEM-EDS), and Transmission Electron Microscopy (TEM) were used to verify that the Se particles produced by L11 are SeNPs with diameters ranging from 50 to 200 nm. The combination of the protein analysis of different cell components and TEM analysis showed that L11 mainly produces SeNPs through the transformation of the cell’s periplasmic space, cell membrane, and cell wall. Adding the L11 SeNPs complex to sheep feed can significantly enhance the antioxidant activity and immunity of sheep, and increase the Se content in the neck muscles, liver, and spleen tissues.
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优化枯草芽孢杆菌菌株 L11 的转化条件以制备纳米硒及其在绵羊饲料中的初步应用
与无机硒相比,纳米硒粒子(SeNPs)具有更高的生物利用率和安全性,被广泛应用于医疗、农业、营养补充剂和抗菌领域。本研究从中国湖北省富硒奶牛养殖基地筛选出一株可产生 SeNPs 的菌株 L11。通过生理、生化和分子生物学分析,确定该菌株为枯草芽孢杆菌。通过调整培养条件,实验确定了 L11 高效生产 SeNPs 的理想参数。实验采用 X 射线光电子能谱(XPS)、扫描电子显微镜-能量色散光谱(SEM-EDS)和透射电子显微镜(TEM)来验证 L11 产生的 Se 粒子是直径为 50 至 200 纳米的 SeNPs。结合不同细胞成分的蛋白质分析和 TEM 分析表明,L11 主要通过改造细胞的周质空间、细胞膜和细胞壁产生 SeNPs。在绵羊饲料中添加 L11 SeNPs 复合物可显著提高绵羊的抗氧化活性和免疫力,增加颈部肌肉、肝脏和脾脏组织中的 Se 含量。
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