Biosynthesis and FPLC purification of antibacterial peptide from the biotherapeutic agent Enterococcus faecium

IF 0.7 Q4 PHARMACOLOGY & PHARMACY
Eslam Abd-elwahed, A. El-Waseif, D. Maany
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引用次数: 1

Abstract

Background Probiotics are microorganisms that play an essential role in microbial intestinal balance and in health care. Objective To isolate a probiotic that can be used to produce antimicrobial peptides potentially used as inhibitors against pathogenic bacteria. Materials and methods The research protocol was carried out through isolation of samples from different dairy product and screening for the most potent probiotic exhibiting antimicrobial activity against Staphylococcus aureus ATCC 6538 and Escherichia coli ATCC 8739 according to the guidelines of Clinical and Laboratory Standards Institute using the disc diffusion method. The molecular identification of this probiotic strain was done by 16S ribosomal DNA sequencing, and the phylogenetic tree was obtained. The purification process and characterization of the antibacterial peptide were done by (NH4)2SO4 and performing fast protein liquid chromatography. Results and discussion Bacterial probiotic strains obtained from different samples were screened for the best antimicrobial activity, where isolate number 9 from 18 isolates showed the highest antibacterial activity against S. aureus and E. coli. Therefore, it was chosen for molecular identification. The molecular identification process revealed that isolate number 9 was Enterococcus faecium. Results of antibiotics sensitivity indicated that S. aureus is more sensitive to antibiotics than E. coli. The fast protein liquid chromatography purification and characterization process of the peptide produced from the probiotic E. faecium showed that the active fraction was precipitated at 60% saturation of (NH4)2SO4. Moreover, single absorbance peaks confirmed the presence of the peptide ‘enterocin.’
粪肠球菌生物治疗剂抗菌肽的生物合成及FPLC纯化
益生菌是在肠道微生物平衡和保健中发挥重要作用的微生物。目的分离一种可用于生产抗菌肽的益生菌,该抗菌肽具有抑制病原菌生长的潜力。材料与方法本研究方案采用圆盘扩散法对不同乳制品样品进行分离,筛选对金黄色葡萄球菌(Staphylococcus aureus) ATCC 6538和大肠杆菌(Escherichia coli) ATCC 8739抗菌活性最强的益生菌,按照临床与实验室标准协会的指南进行。通过16S核糖体DNA测序对该菌株进行分子鉴定,得到系统发育树。采用(NH4)2SO4和快速蛋白液相色谱法对抗菌肽进行纯化和表征。结果与讨论从不同样品中筛选出的益生菌对金黄色葡萄球菌和大肠杆菌的抑菌活性最高,其中18株分离物中9号分离物对金黄色葡萄球菌和大肠杆菌的抑菌活性最高。因此,选择它进行分子鉴定。分子鉴定结果表明,第9号分离物为粪肠球菌。抗生素敏感性结果显示金黄色葡萄球菌对抗生素的敏感性高于大肠杆菌。对益生菌E. faecium产生的肽进行了快速蛋白液相色谱纯化和表征,结果表明,活性部分在60%饱和(NH4)2SO4条件下沉淀。此外,单吸光度峰证实了肽“肠霉素”的存在。
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来源期刊
Egyptian Pharmaceutical Journal
Egyptian Pharmaceutical Journal PHARMACOLOGY & PHARMACY-
CiteScore
1.10
自引率
0.00%
发文量
37
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