N-acetyl transferase purification, What to do next?

Biology mpiraee at hotmail.com
Fri Mar 5 13:33:20 EST 1999


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
Any commnet on how to proceed for further purification is welcomed.

Thanks
Mahmood   mpiraee at is2.dal.ca




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