N-acetyl transferase purification, What to do next?

Achim Recktenwald achimr at home.com
Fri Mar 5 19:26:18 EST 1999

Biology wrote in message <36E02370.324E5276 at hotmail.com>...
>Hi evryone
>It is a long time I am trying to purify N-acetyl transferase from
>Streptomyces venezuelae. It works on a specific substrate p-nitro phenyl
>serinol. I fractionated the crude cell extract by 60% amonium sulfate
>(after removing the pellet fro 30% am. sulfate) which is highly active.
>Then on DEAE collumn it was again fractionated by NaCl gradient. I found
>the active fraction with the highest activity. I analyzed that fraction
>with HPLC. It showed still it has two more proteins with very close MW.
>I already tried HIC columns but it did not worked. I need the peptide to
>be sequenced

Could you mention, why it did not work with HIC?
Which resins did you try?


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