3 replies to this topic

[Puteri Q]

Hi there...
Ive been doing my research about protein overexpression. Is there anyone who knows any factor that may affect to protein expression?
Does the enviromental conditon (ex. medium) may affect to the solubility of the recombinant protein?

P.S.: transforman==> E.coli BL21 DE3 Plyss

--Q--

[perneseblue]

Puteri Q, on Jul 1 2010, 07:26 PM, said:

Hi there...
Ive been doing my research about protein overexpression. Is there anyone who knows any factor that may affect to protein expression?
Does the enviromental conditon (ex. medium) may affect to the solubility of the recombinant protein?

P.S.: transforman==> E.coli BL21 DE3 Plyss

--Q--

Yes, the environment does affect the solubility and expression of recombinant protein. This is why we spend so much money and time optimising growth condition. And it is also the reason why it is so difficult to scale large bio reactors.

Examples of factors that affect protein expression,
Media Carbon-nitrogen ratio.
Carbon source composition.
Nitrogen source composition/availability.
Temperature
Oxygen availability
Waste product build up
Cellular growth rate
pH

It is complicated and i only touched the periphery of this field when I was reading up factors that influence plasmid production and cell growth density.

[Puteri Q]

perneseblue, on Jul 6 2010, 04:22 AM, said:

Puteri Q, on Jul 1 2010, 07:26 PM, said:

Hi there...
Ive been doing my research about protein overexpression. Is there anyone who knows any factor that may affect to protein expression?
Does the enviromental conditon (ex. medium) may affect to the solubility of the recombinant protein?

P.S.: transforman==> E.coli BL21 DE3 Plyss

--Q--

Yes, the environment does affect the solubility and expression of recombinant protein. This is why we spend so much money and time optimising growth condition. And it is also the reason why it is so difficult to scale large bio reactors.

Examples of factors that affect protein expression,
Media Carbon-nitrogen ratio.
Carbon source composition.
Nitrogen source composition/availability.
Temperature
Oxygen availability
Waste product build up
Cellular growth rate
pH

It is complicated and i only touched the periphery of this field when I was reading up factors that influence plasmid production and cell growth density.

Ooo..thanks for the answer.   Do you have any suggestion for the reference that I could learn about those factors?

[perneseblue]

I looked up pubmed and google.

Just start off with a simple inquiry like "protein expression improvement enhancements". That should get you a few papers and reviews. Read them up and you'll pick up more specific terminology and become familiar with the authors who work in this field. Then you can use the correct terminology to improve your search.

And also look up the US patent office. You will probably find most of the more advance/recent work in protein expression enhancements "missing". But by looking at the patent office, you can get a glimpse of what modern companies are doing now. Protein expression is big money.