M9 (probably THE standard chemically defined medium in maniatis) does
not give higher growth than about OD 5 in simple erlenmeyer.
If really maximum volumetric yield is an aim, use a fermenter and
(fed-batch and) rich media or (if not available) just terrific broth
in batch.
best,
David
(PS never heard of induction @ 42, funny system ... we rather like it
cool for making [very functional:] protein)
2009/4/13, megha goyal <mgbiotec from gmail.com>:
> we are currently producing our protein using e.coli w3110 containing puc 18
> plasmid having the gene of interest. our protein is temperature inducible
> i.e we grow our culture at 30º C and induction is at 42 º C. our protein is
> expressed as Inclusion bodies.
>> we are using complex medium having yeast extract and tryptone with addtion
> of trace metals and vitamins during fermentation. glucose is added in
> initital stage as well as fed intermittently as per the requirement mostly
> durign the induction. the culture grows to an O.D. of 10 [6hrs] and then we
> induce it [2hrs] this is as per our technology providers advice.
>> we would like to switch over to chemically defined media from complex media
> so as to increase yield. can anyone suggest how can we go about it as the
> media constituent we found on the net gives trace metals content to be added
> but not of vitamins. kindly help.
>> biotef
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> Methods mailing list
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--
David Minde MSc (TUM)
Cellular Protein Chemistry Room 707
Department of Chemistry
Faculty of Science
Utrecht University
Krytgebouw
Padualaan 8
NL-3584 CH Utrecht
The Netherlands
office phone +31 30 253 4105
mobile phone +31(0)631154267
private address:
Griftkade 4 bis
3572 TW Utrecht
Science is what happens while we are making other plans (~John Lennon)
