Application of Bioassay-Guided Fractionation Coupled with a Molecular Approach for the Dereplication of Antimicrobial Metabolites

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• 作者：Devaraju Rakshith ; Parthasarathy Santosh ; T. P. Pradeep…
• 关键词：Bioprospecting ; Hyphenated techniques ; High ; throughput ; Polyketides ; Polyketide synthase gene ; Phylogenomics ; Dereplication ; Bioautography ; Strain prioritization
• 刊名：Chromatographia
• 出版年：2016
• 出版时间：December 2016
• 年：2016
• 卷：79
• 期：23-24
• 页码：1625-1642
• 全文大小：
• 刊物类别：Chemistry and Materials Science
• 刊物主题：Chromatography; Proteomics; Pharmacy; Laboratory Medicine;
• 出版者：Springer Berlin Heidelberg
• ISSN：1612-1112
• 卷排序：79

文摘

A systematically delineated dereplication approach was described based on genome mining and bioassay-guided fractionation using endophytic fungus Xylaria psidii FPL-52(S) isolated from leaves of Ficus pumila Linn., (Moraceae). A polyketide synthase gene-based molecular screening strategy by a degenerate oligonucleotide primer polymerase chain reaction technique coupled with a bioinformatic phylogenomic approach revealed the presence of an iterative polyketide synthase gene within the genome of Xylaria psidii FPL-52(S). Chemical dereplication of ethyl acetate extract derived from a submerged fermentation culture broth of Xylaria psidii FPL-52(S) by bioassay-guided chromatographic and hyphenated analytical spectroscopic techniques led to the identification of polyketide mycoalexin 3-O-methylmellein. Antimicrobial profiling and minimal inhibitory concentration values for 3-O-methylmellein were determined by disc diffusion and microbroth dilution techniques. Gram-positive bacteria, dermatophytic and phytopathogenic fungi were susceptible in terms of inhibition zone and minimum inhibitory concentration values when compared to co-assayed standards. Herein, we highlight and demonstrate an improved approach which facilitates efficient dereplication and effect-guided fractionation of antimicrobial metabolite(s). The present work flow serves as a promising dereplication tool to survey the biosynthetic potential of endophytic fungal diversity, thereby identifying the most promising strains and prioritizing them for novel polyketide-derived antimicrobial metabolite discovery.