Team:IISER-Pune-India/TeamNotebook/Team Notebook 11e9906b405543a29361e38a50c81024/Meeting with Dr Anand Ghosalkar(Praj) 73814c5603eb47f3b629c704d9ff2fc4

Meeting with Dr Anand Ghosalkar(Praj)

Meeting with Dr Anand Ghosalkar(Praj)

DescriptionIHP on butanol and industry
HP sub-branchiHP
Property 1
Property 2

Project Presentation: Likhith

  1. If our supernatant has equal parts of butanol ethanol, and some pyruvate, do there exist economically viable ways to separate these products. Likhith
  1. Relevance of feedback inhibition and its effect on butanol production. Likhith
  1. If biomass is 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? Sanjana
  1. What do we do with ecoli biomass Sanjana
  1. Immobilisation
    1. how will it impact growth rate, growth dynamics will change
    1. inexpensive and will immobilize the culture
  1. Constitutive vs. inducible promoter
  1. Which industries will be interested in using their emissions? Will, there be other toxic substances in the emissions, can we purify the CO2? (Idk if it's that relevant)
  1. How do we find the optimal frequency of extraction of butanol if we do continuous extraction? What parameters would this depend on?
  1. Will the butanol go back into the cell if it stays in the media for a long time and how much?
  1. Minimum inhibitory concentration - assay
  1. How do we deal with butanol toxicity in cyanobacteria?


green biologics, jule biotechnology

  • butanol vs succinate
    • primary idea lies in utilising co2
    • n-butanol also has equally relevant properties, at least as a fuel
    • high energy density over ethanol and other biofuels, lot of literature on it
    • logistically
    • transporter
      • panels who'd look at metabolic engineering POV
        • rate at which co2, production rates in sucrose
        • he works on downstream systems like ecol and yeast
      • channels a lot to glycogen
        • 8g/l, you can cultivate ecoli in it
        • put up these numbers in your slides
      • been done before?
        • ^combination of symporter and sps gene
        • ^culture works
        • he has experience in methanotrophs
        • have a look at if anyone else has done coculture of cyanobacteria and ecoli before
        • highlight key changes in your project from previous literature
    • invertase enzyme
  • this co2 to butanol in coculture is new, so highlight this
  • scaling up
    • tolerance of ecoli to nbutanol
    • maximum titer, from claustridium, more than 2-3% tolerance not reported
    • separation from fermentation broth might be hard, boiling point higher than water so
    • highlight strategies like extractive fermentation, which solves both toxicity but also simplifies separation of butanol from water
    • pyruvate he hasn't come across
    • claustridium based systems- green biologics
      • from sugar feedstocks ^^
      • separation of ethanol butanol is not much of a problem (distillation)
    • extractive fermentation
    • biocompatible solvent - selected and screened based on toxicity to the microbial system you're using
    • photobioreactor
      • issues in design
        • tubular? lot which have been scaled up for algal have been tubular, 1000 cubic metres has been largest?
        • very rarely in ecoli, they have large bioreactors
        • scale up hasn't been demonstrated very well in last few years
        • jule biotech (might be ded company, he doesn't seem to have contacts there)
          • huge cyanobacterial cultivations, ponds
      • you can get an idea of what's the largest bioreactors possible
      • just highlight that you're aware of what the state of bioreactors is
    • energies
      • rates, titres, of final products
    • highlight that you're solving feedstock problem
      • overall co2 to sucrose to butanol yield - 0.9 * 0.4 - highlight this
    • highlight tolerance of the butanol producing strain, compare with clostridium
    • current molecule is isobutanol, toxicity, yield, ease of engineering
    • praj itself has yeast strain that uses feedstock to butanol?
    • inhibitory feedbacks?
      • clostridia has more literature
      • transcriptomic, proteomic analysis, etc for butanol
      • there was serious work before to produce butanol in clostridia
    • recycling biomass
      • large volume cell biomass
      • cyanobacteria
        • sunlight fixing advantage
        • concern is
          • growth rate
            • report the fast growth rate and its scale
          • photobioreactor scalability
            • led, energy input references should be available in literature?
      • a major source of protein
        • animal feed easiest
        • sellable and acceptable idea
      • antibiotic markers
        • modified organism safety issues?
          • there are
          • DTGS grain-based ethanol plants, GMO corn?
          • say you can remove markerless systems
          • even if is GMO, in the longer run
          • protein limitation problem in the world, 10 billion population
      • immobilization
        • inexpensive?
        • has been around as an idea for v long
        • as of today, people have not been able to utilize it
        • scalability of immobilization is a challenge
        • enzyme catalysed reaction immobilization has been shown
        • highlight dissolved oxygen aspect? missed what likku said
        • would immobilization affect intake rates?
          • non uniform growth becomes challenging when scaled up
          • new innovations
          • immobilized systems for butanol might not have been attempted at all
        • ROS species quenching
          • if cell density is high and it can accumulate peroxide
          • he doesn't see it becoming disadvantaged in an immobilized culture
      • continuous extraction process has been demonstrated before so you can present it as a way:
        • designed in a way to solve toxicity problems and feedback inhibition problems
      • industries for co2 sources:
        • ethanol plants, cleanest co2 which you can get
          • you have advantages of integrating this into ethanol plants
        • onsite production? as a product butanol is good enough
      • extractive fermentation -lots of literature in claustridium for models and values for extraction frequencies
    • tolerance measurement- viability of cells, titration of cell count/solvent concentration, specfici growth rate, productions rates at diff titer values
    • easiest is HPLC, there could be enzymatic methods but he doesn't know for sure, hplc is used routinely in labs, can give precise quantities. GCMS you can also do but he thinks hplc is more common. both would be accurate, sensitive and repeatable.