Abstract
Rosmarinic acid synthase from cell cultures of Coleus blumei Benth. was purified to apparent homogeneity by fractionated ammonium sulfate precipitation (60–80% saturation), hydrophobic interaction chromatography, affinity chromatography and gel filtration. This purification procedure resulted in a 225-fold-enriched specific enzyme activity with a yield of 9%. The protein preparation was apparently pure according to sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and two-dimensional gel electrophoresis. The apparent molecular mass determined by gel filtration and SDS-PAGE was 77 kDa, indicating that rosmarinic acid synthase is a monomeric enzyme.
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Abbreviations
- DTT:
-
dithiothreitol
- HIC:
-
hydrophobic interaction chromatography
- RA:
-
rosmarinic acid
- RAS:
-
rosmarinic acid synthase
- SDS-PAGE:
-
sodium dodecyl sulfate-polyacrylamide gel electrophoresis
References
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The financial support of the Deutsche Forschungsgemeinschaft is gratefully acknowledged. Two-dimensional gel electrophoresis was done with the help of Dr. Guy Bauw, University of Gent, Belgium.
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Petersen, M. Purification of rosmarinic acid synthase from cell cultures of Coleus blumei Benth. Planta 191, 18–22 (1993). https://doi.org/10.1007/BF00240891
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DOI: https://doi.org/10.1007/BF00240891