Abstract
Methods are described for growing the cyanobacterium A. variabilis and for the isolation and purification of plastocyanin from the grown culture. Cell paste which had been stored at −35°C was suspended in 1 mM MES buffer, pH 6.5 and centrifuged. The supernatant was diluted to a conductivity of 0.12 mS, [Fe(CN)6]3- added to a concentration of 0.5 mM and the solution loaded on a S Sepharose Fast Flow column. After elution and ultrafiltration, the plastocyanin containing fractions were reloaded on a S Sepharose Fast Flow column for final purification. A typical yield in three days from cells harvested from 3×20 l of medium was 32 mg plastocyanin with a minimum absorbance ratio A278/A597=1.14. This procedure is faster and the yield higher than for previous procedures.
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Abbreviations
- MES:
-
2(N-morpholino)ethanesulfonic acid
- PC:
-
plastocyanin
References
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Christensen, H.E.M., Conrad, L.S. & Ulstrup, J. A new procedure for fast isolation and purification of plastocyanin from the cyanobacterium Anabaena variabilis . Photosynth Res 25, 73–76 (1990). https://doi.org/10.1007/BF00051737
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DOI: https://doi.org/10.1007/BF00051737