Hi all,
I met a problem about recombinant protein production from yeast CEN.PK_alpha. The target enzyme should secrete into the medium. First, the strains were cultured in 20 mL medium for around 30 h (OD ~ 0.5) and then they were transferred to 200 mL medium with an initial OD of 0.05. When OD reaches 1, the medium was obtained by centrifugation. After ammonium sulfate precipitation overnight, there is almost no pellet on the bottom.
Is there anything wrong here? Could anyone give me some suggestions?
Thank you very much.
Hi Wu,
Before using ammonium sulphate for precipitation, you may need to consider optimising the process by considering different saturated solutions of the salt. Literature search on the enzyme should inform your choice of precipitating temperature and time. You may also need to use ultracentrifugation to harvest the precipitated enzyme. My regards
Try this: may be you find some protocols and booklets.
http://www.biodatacenter.ir
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