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(Reference retrieved automatically from Web of Science through information on FAPESP grant and its corresponding number as mentioned in the publication by the authors.)

Solid phase microextraction as a powerful alternative for screening of secondary metabolites in actinomycetes

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Author(s):
Acquaro Junior, Vinicius Ricardo [1] ; Rodrigues, Julia Pereira [2] ; Beraldo Moraes, Luiz Alberto [2]
Total Authors: 3
Affiliation:
[1] Univ Sao Paulo, Inst Quim Sao Carlos, Sao Paulo - Brazil
[2] Univ Sao Paulo, Fac Filosofia Ciencias & Letras Ribeirao Preto, Dept Quim, Sao Paulo - Brazil
Total Affiliations: 2
Document type: Journal article
Source: Journal of Mass Spectrometry; OCT 2019.
Web of Science Citations: 0
Abstract

Actinobacteria are one of the most promising producers of medically and industrially relevant secondary metabolites. However, screening of such compounds in actinobacteria growth demands simple, fast, and efficient extraction procedures that enable detection and precise quantification of biologically active compounds. In this regard, solid phase microextraction (SPME) emerges as an ideal extraction technique for screening of secondary metabolites in bacteria culture due to its non-exhaustive, minimally invasive, and non-destructive nature: its integrated sample preparation workflow; balanced coverage feature; metabolism quenching capabilities; and superior cleanup, as well as its versatility in configuration, which enables automation and high throughput applications. The current work provides a comparison of micro-scale and direct immersion SPME (DI-SPME) for screening of secondary metabolites, describes the optimization of the developed DI-SPME method, and introduces the developed technique for mapping of target secondary metabolites as well as its direct coupling to mass spectrometry for such applications. The optimized DI-SPME method provided higher amounts of extracted ions and intensity signals, yielding superior extraction and desorption efficiency as compared with micro-scale extraction. Studied compounds presented stability on the coating for 24 h at room temperature. The DI-SPME mapping approach revealed that lysolipin I and the lienomycin analog are distributed along the center and edges of the colony, respectively. Direct coupling of SPME to MS provided a similar ions profile as SPME-LC-MS while enabling a significant decrease in analysis time, demonstrating its suitability for such applications. DI-SPME is herein presented as an alternative to micro-scale extraction for screening of secondary metabolites in actinobacteria solid medium, as well as a feasible alternative to DESI-IMS for mapping of biologic radial distribution of secondary metabolites and cell life cycle studies. Lastly, the direct coupling of DI-SPME to MS is presented as a fast, powerful technique for high throughput analysis of secondary metabolites in this medium. (AU)

FAPESP's process: 16/22023-0 - Activation of the Cryptic Secondary Metabolite Biosynthetic Gene Clusters in Actinomycetes by Antibiotics Resistance
Grantee:Luiz Alberto Beraldo de Moraes
Support type: Regular Research Grants
FAPESP's process: 18/03956-1 - Development and application of the novelty SPME device - Coated Blade Spray
Grantee:Vinicius Ricardo Acquaro Junior
Support type: Scholarships in Brazil - Post-Doctorate