Optimization of Cyclotide extraction from Taraxacum officinale flowers
Master thesis
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http://hdl.handle.net/11250/295186Utgivelsesdato
2015-08-06Metadata
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- Master's theses (KBM) [888]
Sammendrag
A non-destructive protocol was created for extracting, isolating and detecting cyclotides from cultivated T.
Officinale flower heads. Optimal extraction was achieved by maceration for 15 minutes in 50% MeOH and
steeping plant material at 70 °C for 3 hours. Size exclusion chromatography was applied successfully using a
stationary phase with a molecular cut off at 1000-5000 Da yielding a good separation at 280nm. A molar
attenuation threshold was calculated from a protein standard with purified Kalata B1 and used to validate
isolated fractions. An amber colored fraction containing 0.39 mM protein was applied to a 400 MHz NMR to
determine the presence of cyclotides using extraordinary chemical shifts. Kalata B1 was not confirmed but
NMR showed fingerprint similarities to the standard and a signal at -0.1 ppm. This work demonstrates the
viability of the protocol for future use.