Malathy, Amrita, Yazdani (both said don't immobilise, Yazdani because ROS quenching might not happen, why did amrita say this?): Immobilization of ecoli - (reasons: continuous extraction (both Malathy and Dubey said is important - because of feedback inhibition + toxicity), biomass separation (Malathy), growth rate control).
How relevant is feedback inhibition? - praj guy said we don't need to worry too much; Does it happen? How much will it affect butanol production? - R, F - @SANJANA VASANTH@Vidisha Hate
How do we find the optimal frequency of extraction of butanol if we do continuous extraction? Possible parameters this would depend on: MIC for cyano and e.coli, feedback inhibition - R, F - @SANJANA VASANTH@Vidisha Hate
What butanol assay to design to compare butanol production with and without immobilization? - R, F - @AKASH DUTTA@Misaal Bedi
How to immobilize E. coli? (not alginate cuz expensive and imported) - @LIKHITH CHANDRAGIRI@AKASH DUTTAEncapsulation
How will we assay for E.coli growth and population? - F - @AKASH DUTTA@Misaal Bedi
How will sucrose consumption by E.coli change? It'll reduce presumably, what are the implications of that? - F (possible contamination if sucrose use by ecoli reduces?)
Will the butanol go back into the cell if it stays in the media for a long time and how much? - F
Santanu, Malathy: How to reuse biomass and all components of the coculture. @Vidisha Hate@LIKHITH CHANDRAGIRI
If being used as feed for animals, what safety considerations we need to take into account? Do we remove the modifications before using them as feed? Do we remove antibiotic markers or do genomic integration of csc genes? - F
What other ways to use biomass? - R, F - @Vidisha Hate@AKASH DUTTA
What ways to use other components? - R, F - @Vidisha Hate@AKASH DUTTA
Malathy, Dubey, Annamma: Butanol extraction procedure (Dubey and Malathy said it should be continuous because more butanol in the medium will be toxic + feedback inhibition reducing yield) - ask ucm about MIC
Ask NCL peeps and analytical chemists about this + research on your own - R, F - @SANJANA VASANTH@Vidisha Hate
Research what the minimum inhibitory concentration (MIC) of butanol is for E.coli. What are the MIC values for cyano? - R @SANJANA VASANTH@Vidisha Hate
Butanol toxicity for cyano? Will immobilization help? If butanol is toxic to cyano how do we deal with that? - R, F @SANJANA VASANTH@Vidisha Hate
Butanol extraction methods: All 3 R (research other methods too, once we have a base ask someone about the best method) - @Misaal Bedi@AKASH DUTTA
Where will our bioreactor be? Will we have a source of high CO2? What will our light source be? - R
Are we going to reach out to industries to use their emissions? Which industries will be interested in these? Will there be other toxic substances in the emissions, can we purify the CO2? - F
Based on the answers to the above 2 questions, what parameters are important to look at for our bioreactor - F
Microaerobic conditions - R
Malathy: Increasing CO2 uptake capacity of UTEX - @ARYA NARNAPATTI@NAMASIVAYAM SANKAR
How to reduce byproducts? (ethanol, pyruvate, butyrate) - Use Optknock, FSEOP, etc. - R @ARSH SHAIKH@Misaal Bedi
Santanu: Can the E.coli be sustained by exopolysaccharides alone from cyano? (No sucrose release) - R, F - @LIKHITH CHANDRAGIRI
Santanu: Go for constitutive expression (inducible ones have leaky expression) - @LIKHITH CHANDRAGIRI
Should we use inducible promoters for characterizing the coculture? - F
What would be the best inducible promoters from a sustainability perspective? Light inducible etc. - R, F
Nishad: @Misaal Bedi@AKASH DUTTA
Mycobacteria - in static culture, cover media with film of oil to prevent oxygen diffusion and put redox indicator in media, as media becomes more oxygen-deficient indicator goes from blue to colorless - but this works for small culture volumes.
Look at redox indicators to check if the media is anaerobic. lots of reducing agents can be toxic, check which is compatible. - R
How to check if the media is anaerobic/microaerobic? - R
SANJANA VASANTHVidisha Hate
AKASH DUTTA
LIKHITH CHANDRAGIRI
Misaal Bedi
ARYA NARNAPATTI
ASHLI JAIN
Jason Joby
NAMASIVAYAM SANKAR
Soorya Narayan