Hi, Currently i am working on isolation of natural products fromMArine actinobacteria, Recently i found out one of my actinobacteria strain which is very strange, when ever i want to do large culture, my all flasks colour is change, from one another.I use Gause medium and same parent sample for all of flasks, The original colour of strain is Pink, after 3 days of culture, it starts to give pink colour then slowly after some days it become dark pink, Some become dark red, Some light yellow, some dark yellow, some dark brown, some green , even every bottle have a strange colour.I even tried dark and light reactions for that, In dark it didnt changed colour but on HPLC-DAD there was no difference, And on hplc this strain didn't show any peak totally blank. I mostly cultured it from 7 -18 days but no peak on HPLC, i don't know what is the reason are they changing because of heat? or what? the temperature is 28.Is someone have this experience?please guide me for that.
Can I see a culture on Petri dish on ISP2/ISP4. Do check the pH in each flask.. How do you prepare your inoculum and what is the inoculum size?
Respected Mam, Vikineswary,
Thanks for your help but i just cultured it on Gause medium both solid and broth, I didn't used ISP2/ ISP4. and i don't know why in HPLC it does not show any peak, totally blank.
Hi Syed,
My suggestion for this,
1.You must prepare the ISP2 Solid medium and Gause Medium and compare it visually on plate.
2.By seeing your plate and flask am predicting your culture producing some pigment.
3. How you used this culture filtrate for detecting HPLC-DAD. can you explain clearly . If i know the protocol I can give some suggestion to you on this scenario.
Finally you may interact with me through my e-mail : [email protected]
With Regards,
Dr.P.Praveen.
Fmom your plates- top view -three is evidence of sporulation of your culture. Have you viewed it under microscope ( cover slip culture technique). Marine actinomycetes like starch. After SMF was there dense mycelium production. What was the pH? Looking forward to hearing from you via email
pH is the main factor behind different pigment production. Actinobacterial colour tends to change after sporulation and with increase and decrease of pH. My suggestion is to check its colour in ISP-2 and ISP-4 medium.
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