Hello. I am trying to optimize the culture conditions for a bacterial isolate that produces pigment on nutrient agar but not in nutrient broth. I have tried to increase the incubation time for growth, supplement the media with glycerol as well as incubate the bacterial cell pellet with methanol overnight for release of pigment. Can anyone please suggest any other protocol that will help me? Thank you very much
Are you incubating in tubes or flasks? Sometimes when incubating in broth you need to put the culture on a shaker plate to increase oxygen levels and surface area.
Tasha
A source of stress can induce the production of pigments and other secondary metabolites. Do you incubate in the dark? If so, try to provide a light source.
I think is necessary to increase the oxygen concentration in the culture, as Tasha said. You need to use a shaker (and a small amount of liquid in a large flask) or an air pump and a sterile filter. The temperature is important too.
Dear Prerana,
You can use Orbital shaking Incubator for your purpose. The instrument can provide your culture a suitable incubation period as well, at the same the shaking activity would spread the microbial metabolite production.
Hi, can you infrom that which type of pigment do you want to synthesize?
for your information, use mineral salt media 50mM strength and add the primery metabolites along with nitrogen and carbon source which organism can utilize for the production of your desired pigment (pH7.2 @ 35oC)... I think this information may helpful for your research
As Sajjan told first target the metabolic pathway of the pigment of interest.use a minimal medium like Kurian et al,2014. Use no primary metabolite to create stress..add the precursor of the pigment as the only source of metabolite.
For example in my medium mentioned above I had added only tyrosine as the sole carbon source for melanin production.like wise you can try.
Thank you all for your valuable suggestions. I am incubating the culture in flasks with a light source and in shaking condition. I tried culturing the bacteria in a 250 ml flask with only 50 ml media that resulted in no pigment production. I am currently trying to restrict the bacterial movement by increasing the viscosity of the medium and hoping to give a positive result. Thank you all once again for your valuable suggestions.
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