I am working with Bacillus subtilis RIk1285 (derivative of 168 W.T from TaKaRa), the maximum O.D I get when cultured in L.B broth (Flask work) is 4 O.D and in 7.5 Litre bench top fermentor (Eppendorf) is 5 O.D , apart from LB I did try with SR medium (Peptone; Yeast extract; K2HPO4) which didnt showed any much changes.
Kindly suggest few medium composition or any suggestions to increase the cell density above 10 O.D.
In my opinion, you should concentrate on why OD is not reaching till your expected OD rather than media composition. Perhaps you might be using batch culture in a big fermented. Try continuous culture system. If you do not have that. Try feed batch culture system. You can grow cell as usual in batch fermented. Then filter out the cells, remove used media, then refill fresh media. Then check if it is possible to attend your expected OD. Perhaps metabolite in the media has to do with cell cycle or nutrient deficiency or density dependent growth inhibition.
Dear Goutham,
try a minimal medium:
M9: http://subtiwiki.uni-goettingen.de/wiki/index.php/M9_minimal_medium
CM: http://subtiwiki.uni-goettingen.de/wiki/index.php/C_minimal_medium
You should easily reach OD5-7 in shake flasks using 5 g/L of glucose. By increasing the glucose, ammonium and Mg concentration you should get optical densities higher than 10. Next to glucose, malate is a preferred C-source of Bacillus subtilis. If you want to get higher, I recommend fed-batch culture.
https://www.ncbi.nlm.nih.gov/pubmed/18601168
Good luck
Dear Michael,
I did try in past with M9 medium but i found a very poor growth of my strain in it. I shall try with CM medium once including glucose or any of the above cocktails you mentioned. Thank U :)
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