Explorative solid-phase extraction (E-SPE) for accelerated microbial natural product discovery, dereplication, and purification.
Microbial natural products (NP) cover a high chemical diversity, and in consequence extracts from microorganisms are often complex to analyze and purify. A distribution analysis of calculated pK(a) values from the 34390 records in Antibase2008 revealed that within pH 2-11, 44% of all included compounds had an acidic functionality, 17% a basic functionality, and 9% both. This showed a great potential for using ion-exchange chromatography as an integral part of the separation procedure, orthogonal to the classic reversed-phase strategy. Thus, we investigated the use of an "explorative solid-phase extraction" (E-SPE) protocol using SAX, Oasis MAX, SCX, and LH-20 columns for targeted exploitation of chemical functionalities. E-SPE provides a minimum of fractions (15) for chemical and biological analyses and implicates development into a preparative scale methodology. Overall, this allows fast extract prioritization, easier dereplication, mapping of biological activities, and formulation of a purification strategy.
Center for Microbial Biotechnology, Institute for Systems Biology, Technical University of Denmark, Søltofts Plads, Building 221, DK-2800 Kgs. Lyngby, Denmark. maj@bio.dtu.dk
Maria
M
Månsson
Richard K
RK
Phipps
Lone
L
Gram
Murray H G
MH
Munro
Thomas O
TO
Larsen
Kristian F
KF
Nielsen
D001419:Bacteria / Q000737:chemistry / Q000254:growth & development; D001688:Biological Products / Q000032:analysis / Q000737:chemistry / Q000191:economics / Q000302:isolation & purification* / Q000494:pharmacology; D002852:Chromatography, Ion Exchange / Q000379:methods; D016208:Databases, Factual; D006863:Hydrogen-Ion Concentration; D015394:Molecular Structure; D052616:Solid Phase Extraction / Q000379:methods*
D001688:Biological Products