Team:IISER-Pune-India/TeamNotebook/Team Notebook 11e9906b405543a29361e38a50c81024/Meeting with Dr Santanu Das (Reliance) 579da5145647493c83bb5d44b8f6db62

Meeting with Dr. Santanu Das (Reliance)

Meeting with Dr. Santanu Das (Reliance)

DescriptionIHP on biosafety, scaling up
HP sub-branchiHP
Property 1
Property 2

Meeting Notes

  • explained proj in a few secs
  • You could get sugar from waste, why are you using cyanobacteria?
  • why two organisms
    • cyano is being used as bioengineering by several people
    • if you say it's difficult to engineer
    • it shouldnt be fancy, it should be reasonable
      • minimal modifications
    • many other sources, that could by used by ecoli to produce your molecule
    • challenges of which strain of cyanobacteria
    • same media for ecoli would be needed
    • coculture problems
    • environment: photobioreactor
      • there could be chunks of other organisms
    • if someone chooses to do something else wherein they breakdown the sugar in the medium for
    • some cyanobacteria which in other stresses excrete huge amount of exopolysaccharide
    • how do you use cyanobacteria and ecoli biomass
    • not just molecule of interest
    • geno-waste?
    • what do you do with the rest of the biomass?
      • energy
      • for feed
    • even if your project is on the molecule it's good to show the use for your biomass
    • lots of accumulation in the form of glycogen etc, which might not be useful
    • turbido-stat
      • grow culture and continuously harvest
      • have 100 liters, take 10l away everyday and add same amount of new media
      • water and biomass will be left out, figure out what to do with it
      • recycling, or if water stream has lots of other organic components, we could use anaerobic digestion of the stream to produce methane, or used for cattlefield
      • biomass= combination of biomolecules
      • Can we get just the biomolecules out of the culture?
        • difficult process
    • grow everything for two days, take everything and then reenoculation
    • when the OD goes to something like 0.9, take 1 litre and add new 1 litre
    • 1) proof of concept: few litres
    • 2) pre pilot: testing in 100 litres
    • toxicological and safety trials
    • regulatory approval
    • 3) pilot in 500 litres
    • 4) commercial : replicate in 100 fermentors
    • you don't have to do all this, but we should plan this out and give a complete package
    • antibiotic markers - for food, is a question
    • if the molecule is just a chemical, antibiotic markers are fine
    • breach of containment could be a slight issue if it's possible to be proliferated in nature
    • antiobiotic marker less is only positive for public perception
    • photobioreactor
      • give artificial light
        • good for proof of concept
        • but economics
      • kept in sunlight
        • much more careful
        • while adding, removing media, or clean it, risk of environment exposure
      • we could do it such that for proof of concept we do it in artificial light and then later design it in sunlight
      • light shading and bioreactor would be a project in itself
      • better to use a commercially available bioreactor
      • making it carbon neutral including renewable power sources
      • would be easy enough to add to the project proposal, (renewable sources of energy), hard IRL
    • constitutive or inducible
      • dont think about inducible unless you need fine control, lots of other difficulties with using them
      • constitutive is best option
      • expression level, risk
    • butanol
      • if you're producing too much, cells may not grow
      • first grow it and then induce after certain cell density
    • when you want to express the molecule, when you express the gene, the origin species of the gene - codon optimise, untranslated leader, specific information
    • even in industry it could be both constitutive or inducable
    • very minimal contribution to carbon sequestration
    • to get a scale which makes a difference to industry
    • it might help more in economics to products which you're making than to the environment
    • lanzatech
    • he didnt give any exact numbers regarding how much profit companies would be willing to compromise
    • what questions do you want to ask
    • lay out a plan to let people know
      • complete flowsheet of the fluxes
      • parameters that we should know
    • praj industries
    • carbon capture and storage:

Questions we can ask:

So these questions are more general , not based on his specifications(ask these towards the end ig):

  1. Our two options for end products are Butanol/succinate. So we are aiming at a sustainable, but also economically viable process. Based of the market, current production and uses of both these products, which of these products would be better in terms of both economics and sustainability.
  1. The cyanobacteria that we are working with work best at 3% CO2, whereas in air it is about 0.05%. This got us to think maybe we could use emissions or something from some industries that will have higher CO2 percentage as input for our bioreactor. This will be helpful for us in terms of productivity and increasing incentive, and for the industries to bring down their emissions. Based on this:
    1. Which industries would suit this best
    1. How do we go about approaching this
    1. What are the things we should keep in mind for this

Based on his expertise:

  1. based on - What are things we should keep in mind now for scaling up, as in what alter challenges can be mitigated by work now? How do we generalise a lab culture to industrial setting? What should we look out for? How do we simulate photobioreactor conditions in the lab, how do we create a prototype? Have you seen co-culture being scaled up industrially? How long would a co-culture have to last for it to be industrially relevant?
  1. Are there some interesting strategies(similar to things like Q2 from before) or something he can think of for scaling this up. Things like factory emissions, those make-fuel-at home small bioreactors, etc
  1. Are there things to keep in mind for end-product users ( in terms of purity and all)
  1. based on - bio safety questions essentially, specifically wrt to cyano.

(He is a member of the global future council on synthetic biology), so he would have an idea about 1. current research in synbio and also 2. peoples opinions and the challenges in the implementation of GMO stuff.

5. Would the general public or other industries opinion on synbio affect the implementation of our project? If yes, is there anything we can change or so in our project that can make this better?

6. Are there currently other synbio projects or tools being developed that would compliment our project or something? idk

7.''Don't focus just on making the project sustainable because economically that is useless and not realistic (ease of execution and economic viability is given max importance)" - Dr Odaneth(DBT-ICT)

What is his opinion on this and how do we ensure easy execution and economic viability in our idea?

8. Antibiotics, markers, containment strategies, feasibility for open air culture, cell shading issues

9. possibility of co-culture crashes

10. night time productivity

11. constitutive vs inducible expression, cost of IPTG induction, cost of salt induction, effect of salt on downstream processing

12. geographical conditions needed for an industrial bioreactor set up