Rahman Lubna, Shinwari Zabta K, Iqrar Irum, Rahman Lutfur, Tanveer Faouzia
Molecular Systematics and Applied Ethno Botany Lab (MoSEL), Department of Biotechnology, Quaid I Azam University, Islamabad, Pakistan.
Pakistan Academy of Sciences, 3-Constitution Avenue Sector G-5/2, Islamabad, Pakistan.
Ann Clin Microbiol Antimicrob. 2017 Aug 1;16(1):53. doi: 10.1186/s12941-017-0228-7.
Natural products of animals, plants and microbes are potential source of important chemical compounds, with diverse applications including therapeutics. Endophytic bacteria that are especially associated with medicinal plants presents a reservoir of therapeutic compounds. Fagonia indica has been recently investigated by numerous researchers because of its striking therapeutic potential especially in cancer. It is also reported that endophytes play a vital role in the biosynthesis of various metabolites; therefore we believe that endophytes associated with F. indica are of crucial importance in this regard. The present study aims successful isolation, molecular identification of endophytic bacteria and their screening for bioactive metabolites quantification and in vitro pharmacological activities.
16S rRNA gene sequencing was used for the identification of isolated endophytic bacteria. Methanolic extracts were evaluated for total phenolic contents (TPC), total flavonoids contents (TFC), DPPH free radical scavenging activity, reducing power and total anti-oxidant assays were performed. And also screened for antibacterial and antifungal activities by disc diffusion method and their MIC were calculated by broth dilution method using microplate reader. Further, standard protocols were followed for antileishmanial activity and protein kinase inhibition. Analysis and statistics were performed using SPSS, Table curve and Origin 8.5 for graphs.
Bacterial strains belonging to various genera (Bacillus, Enterobacter, Pantoea, Erwinia and Stenotrophomonas) were isolated and identified. Total phenolic contents and total flavonoids contents varies among all the bacterial extracts respectively in which Bacillus subtilis showed high phenolic contents 243 µg/mg of gallic acid equivalents (GAE) and Stenotrophomonas maltophilia showed high flavonoids contents 15.9 µg/mg quercitin equivalents (QA), total antioxidant capacity (TAC) 37.6 µg/mg of extract, reducing power (RP) 206 µg/mg of extract and 2, 2-diphenyl-1-picrylhydrazyl (DPPH) free radical scavenging activity with 98.7 μg/mL IC value. Although all the extracts tested were active to inhibit growth of selected pathogenic microbes (bacteria and fungi), but significant antibacterial activity was observed against Klebsiella pneumonia and B. subtilis. An Enterobacter cloaca was active against Leishmania tropica with IC value of 1.4 µg/mg extracts. B. subtilis and Bacillus tequilensis correspondingly exhibit significant protein kinase inhibition of 47 ± 0.72 and 42 ± 1.21 mm bald zones, indicating anti-infective and antitumor potential.
Our findings revealed that crude extracts of selected endophytic bacteria from F. indica possess excellent biological activities indicating their potential as an important source of antibiotics (antifungal, antibacterial) compounds.
动物、植物和微生物的天然产物是重要化合物的潜在来源,具有包括治疗学在内的多种应用。与药用植物特别相关的内生细菌是治疗化合物的一个储存库。由于其显著的治疗潜力,尤其是在癌症治疗方面,印度法贡草最近受到了众多研究人员的调查。也有报道称内生菌在各种代谢物的生物合成中起着至关重要的作用;因此,我们认为与印度法贡草相关的内生菌在这方面至关重要。本研究旨在成功分离、分子鉴定内生细菌,并对其进行生物活性代谢物定量筛选和体外药理活性研究。
采用16S rRNA基因测序法对分离出的内生细菌进行鉴定。对甲醇提取物进行总酚含量(TPC)、总黄酮含量(TFC)、DPPH自由基清除活性、还原力和总抗氧化剂测定。并采用纸片扩散法筛选其抗菌和抗真菌活性,用酶标仪通过肉汤稀释法计算其最低抑菌浓度(MIC)。此外,遵循标准方案进行抗利什曼原虫活性和蛋白激酶抑制研究。使用SPSS、Table curve和Origin 8.5进行分析和统计以绘制图表。
分离并鉴定了属于不同属(芽孢杆菌属、肠杆菌属、泛菌属、欧文氏菌属和嗜麦芽窄食单胞菌属)的细菌菌株。所有细菌提取物中的总酚含量和总黄酮含量各不相同,其中枯草芽孢杆菌的总酚含量最高,为243μg/mg没食子酸当量(GAE),嗜麦芽窄食单胞菌的总黄酮含量最高,为15.9μg/mg槲皮素当量(QA),总抗氧化能力(TAC)为37.6μg/mg提取物,还原力(RP)为206μg/mg提取物,2,2-二苯基-1-苦基肼(DPPH)自由基清除活性的IC值为98.7μg/mL。尽管所有测试提取物均能有效抑制所选致病微生物(细菌和真菌)的生长,但对肺炎克雷伯菌和枯草芽孢杆菌观察到显著的抗菌活性。一株阴沟肠杆菌对热带利什曼原虫有活性,提取物的IC值为1.4μg/mg。枯草芽孢杆菌和龙舌兰芽孢杆菌相应地表现出显著的蛋白激酶抑制作用,秃斑区分别为47±0.72和42±1.21mm,表明具有抗感染和抗肿瘤潜力。
我们的研究结果表明,从印度法贡草中筛选出的内生细菌粗提物具有优异的生物活性,表明它们作为抗生素(抗真菌、抗菌)化合物的重要来源具有潜力。