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从本土猪肠道来源的细菌中分离和鉴定一种新型抗菌肽。

Isolation and characterisation of a novel antibacterial peptide from a native swine intestinal tract-derived bacterium.

机构信息

College of Animal Science and Technology, Northwest A&F University, 22 Xinong Road, Yangling, Shaanxi 712100, China.

College of Animal Science and Technology, Northwest A&F University, 22 Xinong Road, Yangling, Shaanxi 712100, China; Institute of Animal Sciences of CAAS, No. 2 Yuanmingyuan West Road, Haidian, Beijing 100193, China.

出版信息

Int J Antimicrob Agents. 2017 Apr;49(4):427-436. doi: 10.1016/j.ijantimicag.2016.12.012. Epub 2017 Feb 27.

Abstract

Antimicrobial peptides (AMPs) are highly associated with antipathogenic activity, without generating drug resistance in targeted bacteria. In this study, the existence of AMPs in the Tibetan swine, a China-native, cold-resistant and seldom-sick breed of pig, was investigated. A peptide secreted by a Tibetan swine intestinal tract-derived Bacillus strain was isolated using reversed-phase chromatography (RPC), ultrafiltration and reversed-phase high-performance liquid chromatography (RP-HPLC). The peptide was identified by mass spectrometry and was characterised for activity against Escherichia coli and Staphylococcus aureus. The 16-amino acid peptide (ASVVNKLTGGVAGLLK), named TP, had a molecular mass of 1568.919 Da and exhibited inhibitory activity against Gram-positive and Gram-negative bacteria [minimum inhibitory concentrations (MICs) of 2.5-5 µM and 10-20 µM for E. coli and S. aureus, respectively] as well as human MKN-45 and NB4 tumour cell lines [50% inhibitory concentration (IC) = 4.686 µM and 11.479 µM, respectively]. TP also exhibited weak haemolytic activity. Furthermore, TP enhanced cell membrane permeability and K outflow, bound with E. coli genomic DNA in vitro and inhibited E. coli growth. Thus, TP represents a strong candidate as an antibacterial peptide.

摘要

抗菌肽(AMPs)与抗病原体活性高度相关,而不会在目标细菌中产生耐药性。本研究调查了中国本土的耐寒、少病的藏猪是否存在 AMPs。使用反相色谱(RPC)、超滤和反相高效液相色谱(RP-HPLC)从藏猪肠道衍生的芽孢杆菌菌株中分离出一种由肽。该肽通过质谱鉴定,并针对大肠杆菌和金黄色葡萄球菌的活性进行了表征。该 16 个氨基酸肽(ASVVNKLTGGVAGLLK),命名为 TP,分子量为 1568.919 Da,对革兰氏阳性和革兰氏阴性细菌均具有抑制活性(对大肠杆菌和金黄色葡萄球菌的最小抑菌浓度(MIC)分别为 2.5-5 μM 和 10-20 μM),以及人 MKN-45 和 NB4 肿瘤细胞系[50%抑制浓度(IC)= 4.686 μM 和 11.479 μM]。TP 还表现出较弱的溶血活性。此外,TP 增强了细胞膜通透性和 K 外流,与大肠杆菌基因组 DNA 在体外结合,并抑制大肠杆菌生长。因此,TP 是一种有前途的抗菌肽候选物。

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