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利用超高效液相色谱-电喷雾串联质谱(UPLC-ESI-MS/MS)和热脱附-气相色谱-质谱联用(TD-GC-MS)分析法,对紫茉莉内生真菌棒曲霉MJ31菌株抗菌化合物的测定及生产

Determination and production of antimicrobial compounds by Aspergillus clavatonanicus strain MJ31, an endophytic fungus from Mirabilis jalapa L. using UPLC-ESI-MS/MS and TD-GC-MS analysis.

作者信息

Mishra Vineet Kumar, Passari Ajit Kumar, Chandra Preeti, Leo Vincent Vineeth, Kumar Brijesh, Uthandi Sivakumar, Thankappan Sugitha, Gupta Vijai Kumar, Singh Bhim Pratap

机构信息

Molecular Microbiology and Systematics Laboratory, Department of Biotechnology, Aizawl, Mizoram University, Mizoram, India.

SAIF, CSIR-Central Drug Research Institute (CSIR-CDRI), Lucknow, India.

出版信息

PLoS One. 2017 Oct 19;12(10):e0186234. doi: 10.1371/journal.pone.0186234. eCollection 2017.

Abstract

Endophytic fungi associated with medicinal plants are reported as potent producers of diverse classes of secondary metabolites. In the present study, an endophytic fungi, Aspergillus clavatonanicus strain MJ31, exhibiting significant antimicrobial activity was isolated from roots of Mirabilis jalapa L., was identified by sequencing three nuclear genes i.e. internal transcribed spacers ribosomal RNA (ITS rRNA), 28S ribosomal RNA (28S rRNA) and translation elongation factor 1- alpha (EF 1α). Ethyl acetate extract of strain MJ31displayed significant antimicrobial potential against Bacillus subtilis, followed by Micrococccus luteus and Staphylococcus aureus with minimum inhibitory concentrations (MIC) of 0.078, 0.156 and 0.312 mg/ml respectively. In addition, the strain was evaluated for its ability to synthesize bioactive compounds by the amplification of polyketide synthase (PKS) and non ribosomal peptide synthetase (NRPS) genes. Further, seven antibiotics (miconazole, ketoconazole, fluconazole, ampicillin, streptomycin, chloramphenicol, and rifampicin) were detected and quantified using UPLC-ESI-MS/MS. Additionally, thermal desorption-gas chromatography mass spectrometry (TD-GC-MS) analysis of strain MJ31 showed the presence of 28 volatile compounds. This is the first report on A. clavatonanicus as an endophyte obtained from M. jalapa. We conclude that A. clavatonanicus strain MJ31 has prolific antimicrobial potential against both plant and human pathogens and can be exploited for the discovery of new antimicrobial compounds and could be an alternate source for the production of secondary metabolites.

摘要

据报道,与药用植物相关的内生真菌是多种次生代谢产物的有效生产者。在本研究中,从紫茉莉根部分离出一株具有显著抗菌活性的内生真菌——棒曲霉MJ31菌株,通过对三个核基因即核糖体RNA内部转录间隔区(ITS rRNA)、28S核糖体RNA(28S rRNA)和翻译延伸因子1-α(EF 1α)进行测序来鉴定该菌株。MJ31菌株的乙酸乙酯提取物对枯草芽孢杆菌显示出显著的抗菌潜力,其次是藤黄微球菌和金黄色葡萄球菌,最低抑菌浓度(MIC)分别为0.078、0.156和0.312 mg/ml。此外,通过扩增聚酮合酶(PKS)和非核糖体肽合成酶(NRPS)基因来评估该菌株合成生物活性化合物的能力。进一步地,使用超高效液相色谱-电喷雾串联质谱(UPLC-ESI-MS/MS)检测并定量了七种抗生素(咪康唑、酮康唑、氟康唑、氨苄青霉素、链霉素、氯霉素和利福平)。此外,对MJ31菌株的热脱附-气相色谱质谱联用(TD-GC-MS)分析表明存在28种挥发性化合物。这是关于从紫茉莉中获得的棒曲霉作为内生菌的首次报道。我们得出结论,棒曲霉MJ31菌株对植物和人类病原体均具有丰富的抗菌潜力,可用于发现新的抗菌化合物,并且可能是次生代谢产物生产的替代来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/ae35/5648158/fe51f81c53b9/pone.0186234.g001.jpg

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