I need high density large volume inoculum for fermentation. Inoculum density of Pesudomonas aeroginosa is at OD600=1.9, in 25 mL LB in 250 mL flask, while it is 1.6 in 50 mL medium in 250 mL flask. Then it drops down to OD600=0.8 in 100 mL medium in 500 mL flask. I increased the no of loops as well, but same result was obtained. (37 degree Celsius/150 rpm)
Check https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5812950/
https://sfamjournals.onlinelibrary.wiley.com/doi/full/10.1111/1751-7915.13905
Also check https://www.tandfonline.com/doi/pdf/10.1080/00986445.2021.2012462
https://www.tandfonline.com/doi/full/10.1080/00986445.2021.2012462
Also check on this paper
https://www.mdpi.com/2227-9717/10/2/307/pdf
What is the composition of cultivation medium and what is the substrate and what is its concentration?
Regards
Vit
Dear Kousar Perveen:
I have been in the fermentation industry for over 35 years and I never use 25 ml/50 ml medium in 250 ml conical flasks. The general practice is 50 ml medium in 500 ml flask is the standard configuration or you may use wide mouth 500 ml conical flasks. If the shaker through is about 50 mm then it is better and if the shaker through is 25 mm then you may have to increase the rpm to 200 - 225 rpm. You may also baffle flask which is available or you may have to make it yourself or with the help of a glass blower.
Pseudomonas aeruginosa is a highly aerobic bacterium. Therefore you must consider increasing aeration with increased volume. If it exhausts the available oxygen in the medium, its growth rate will drop and inoculum density will drop as well. If it is a strain that can switch from oxidative metabolism to anaerobic respiration, still the relevant external electron acceptor must be available. As the bacterium is metabolically versatile, care must be taken to not overburden it with garbage in the medium.
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