Hi
Someone can help me
How can I increase the cell biomass of Bacillus coagulans and then stimulate sporulation. I use a 10-liter fermenter to coagulans culture. The basic culture medium consists of csl and salts. Please tell me the conditions of cultivation and the amount and type of salts used (most important salts) for s coagulans cultivation.
Dear Amin,
when you need more biomass you can try high-cell density fed-batch cultivation to reach high ODs. Just feed in glucose after carbon supply during batch growth is depleted. Maybe add some nitrogen and Mg to the feeding solution.
In order to trigger sporulation just let the cells starve into carbon limitation. You may also add/activate a stressor that causes sporulation in bacilli (heat, high/low pH, oxidative stress). Or you may switch to a sporulation medium.
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Good luck
Michael
Hello, use reach liquid media with rotating or shaking better with aeration. To simulate sporulation after grow put culture in refrigerator 2-4 *C for 5-7 days.
Michael Kohlstedt
Thanks for your guide Dear Michael Kohlstedt. Can I use high temperature and mncl2 salt for sporulation simultaneously? What temperature can I use for sporulation? And how long should the process be at that temperature ?
I am looking for a quick and practical way to do this. Because in our company, the time of coagulans fermentation process is 40 hours!
Try a 30 min heat shock at 42-45°C. In order to check sporulation efficiency take a culture sample after the heat shock, plate out different dilutions of the sample, put the dilutions to 80°C for 30 min and plate them out again. Compare the colony number before and after heat treatment to see how many cells underwent sporulation in your cultivation.
Check published sporulation media for various Bacillus species:
https://openwetware.org/wiki/Cfrench:Sporulation
https://www.dsmz.de/microorganisms/medium/pdf/DSMZ_Medium531.pdf
M
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