Preparation of microbial broth and plant extract libraries and search for interesting secondary metabolites

　　　Contact: Akiko Okano, Nobumoto Watanabe
　　　E-mail: tech_CRAST

Abstract

　Microorganisms, such as actinomycetes and fungi, have produced wide variety of secondary metabolites in structure and biological activity. Such metabolites have been used as drugs, agrochemicals, and their lead compounds. In a chemical biology study, they are used as a bioprobe, which is a chemical tool to investigate biological functions and an important key. Therefore, we have been preparing a microbial broth library from microbial strains by isolating actinomycetes and fungi from soils collected in various region of Japan. The library is used as a unique screening source for interesting biological activity tests. Also, physicochemical information including UV and mass spectral data obtained by LC/MS analysis is used for searching structurally novel metabolites. Plants including medicinal plants and vegetables, which also contain various biologically interesting compounds, have been used as a source to prepare a plant extract library.


Preparation of a microbial broth library

  1. 1. Collection of soils
  2. 2. Isolation of microbes from soils
  3. 3. Culture of a microbial strain by some culturing conditions
  4. 4. Preparation of an extract by solvent extraction and drying up
  5. 5. Preparation of a library
  6. 6. Validation of a library by LC/MS
  7. 7. Supply as a screening source for bioassays

Preparation of a plant extract library

  1. 1. Collection of plants
  2. 2. Preparation of extraction: cleaning and drying
  3. 3. Extraction with organic solvent and dry up
  4. 4. Liquid-liquid partition between organic solvent and water
  5. 5. Preparation of a library
  6. 6. Validation of a library by LC/MS
  7. 7. Supply as a screening source for bioassays



Overview of preparation of a microbial broth library

Search for interesting metabolites

Bioassay guided separation

　We are searching for biologically interesting metabolites based on various biological assays, such as antimicrobial activity and cytotoxicity against mammalian cells.


Structure oriented separation

　We are searching for structurally interesting metabolites based on DAD-LC/MS data including UV and mass spectra.




Search for new metabolites based on LC/MS data

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