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内生天蓝色链霉菌菌株AZRA 37抗菌特性的表观遗传激活及利用基质辅助激光解吸电离飞行时间串联质谱鉴定诱导蛋白

Epigenetic Activation of Antibacterial Property of an Endophytic Streptomyces coelicolor Strain AZRA 37 and Identification of the Induced Protein Using MALDI TOF MS/MS.

作者信息

Kumar Jitendra, Sharma Vijay K, Singh Dheeraj K, Mishra Ashish, Gond Surendra K, Verma Satish K, Kumar Anuj, Kharwar Ravindra Nath

机构信息

Mycopathology and Microbial Technology Laboratory, Department of Botany, Banaras Hindu University, Varanasi, 221005, India.

Dept of Botany, Visva-Bharati, Shantiniketan, Bolpur, India.

出版信息

PLoS One. 2016 Feb 4;11(2):e0147876. doi: 10.1371/journal.pone.0147876. eCollection 2016.

Abstract

The endophytic Streptomyces coelicolor strain AZRA 37 was isolated from the surface sterilized root of Azadirachta indica A. Juss., commonly known as neem plant in India. Since only a few reports are available regarding epigenetic modulations of microbial entities, S. coelicolor was treated with different concentrations of 5-azacytidine for this purpose and evaluated for its antibacterial potential against five human pathogenic bacteria (Aeromonas hydrophila IMS/GN11, Enterococcus faecalis IMS/GN7, Salmonella typhi MTCC 3216, Shigella flexneri ATCC 12022 and Staphylococcus aureus ATCC 25923). The crude extract obtained from cultures treated with 25 μM concentration of 5-azacytidine, was found effective against all five pathogenic bacteria tested while the untreated control was only active against 3 pathogenic bacteria. HPLC analysis of crude compounds from treated cultures showed a greater number of compounds than that of the control. Extraction of whole cell protein and its SDS PAGE analysis showed an additional major protein band in 25 μM 5-azacytidine treated culture and MALDI TOF MS/MS analysis revealed that this protein belongs to the porin family.

摘要

内生链霉菌天蓝色链霉菌菌株AZRA 37是从印度楝(在印度通常称为印楝树)经表面消毒的根部分离得到的。由于关于微生物实体表观遗传调控的报道较少,因此为此用不同浓度的5-氮杂胞苷处理天蓝色链霉菌,并评估其对五种人类病原菌(嗜水气单胞菌IMS/GN11、粪肠球菌IMS/GN7、伤寒沙门氏菌MTCC 3216、福氏志贺氏菌ATCC 12022和金黄色葡萄球菌ATCC 25923)的抗菌潜力。从用25μM浓度的5-氮杂胞苷处理的培养物中获得的粗提物对所有五种测试病原菌均有效,而未处理的对照仅对3种病原菌有活性。对处理过的培养物中的粗化合物进行HPLC分析表明,其化合物数量比对照更多。全细胞蛋白的提取及其SDS-PAGE分析显示,在25μM 5-氮杂胞苷处理的培养物中有一条额外的主要蛋白带,MALDI TOF MS/MS分析表明该蛋白属于孔蛋白家族。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/8b6b/4742224/ef89fd4cc7b9/pone.0147876.g001.jpg

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