Green Pacific Biologicals
              Junio 2012
    Leandro Vetcher – CEO

   lvetcher@greenpacificbio.com
            @lvetcher


          Green Pacific Biologicals - Not for distribution
GPB: Plataforma de ingenieria genetica




                                        GPB
 Stable, stronger                      More efficient                                Access to the
 gene expression                        engineering                                   entire cell




                                        other
                                                  0
                          50

                                  transformants
                                         50
               % expressing       % expressing
               transformants                      other          GPB
                                            0



            Only company with rapid and stable
        eukaryotic algae nuclear genetic engineering
         Green Pacific Biologicals - PROPRIETARY and CONFIDENTIAL - Not for distribution         2
GPB: Soluciones para biocombustibles

        Increase carbon
            fixa on



                                                                             Implement
                                                                               efficient
                    Maximize                                                 oil secre on
                   carbon flux
                     into oil




       Green Pacific Biologicals - PROPRIETARY and CONFIDENTIAL - Not for distribution      3
Expresión génica nuclear mas fuerte

 GFP expression by Western blotting                 Accumulation of GFP by confocal microscopy



 GFP protein         Cell extract



                                      Wild-type
  stds (ng)           WT GPB


                                      GPB Strains




                                                                                          4
Avg. fluorescence
           per cell (au)




    Avg. fluorescence
      per cell (au)
                             Expresión génica nuclear estable




5
Mas transformantes expresando transgenes

                      More rapid and efficient
                           engineering




       Green Pacific Biologicals - PROPRIETARY and CONFIDENTIAL - Not for distribution   6
RNAi funcional- ingeniería metabólica
     Genetic RNAi using GPB platform better than
                 traditional mutant

  Starch lower      Oil higher      Growth normal




                                                   7
Localización subcelular de proteína
                         Membrane             Lipid droplet
                         localization          localization




 Untransformed cell   Membrane part+GFP Lipid droplet part+GFP



                                                              8
Secreción de aceites no-toxica
 Non-toxic mechanism
 Secreted LDs spontaneously separates, sidestepping
  harvesting/extraction
 Works by interactions between proteins on LDs and cell membrane
  or other cellular components
 Platform makes GPB uniquely positioned to implement oil secretion
  in algae




                                                                9
Beneficios de secreción de aceites no-toxica
BASE CASE:                          GPB CASE:
• Milk harvested by killing cow     • Milk harvested through secretion and cow returned to pasture

• Majority of energy  Making cow   • Majority of energy (grass) goes into making milk

                                    • If cows are milked daily, there is no benefit to having cows that can
                                    store more than one day’s worth of milk


MILK      +                                             MILK




                                                                                     ?
Modelado Tecno-económico
      Oil productivity model + operational & capital costs
• Physics: energy balanced,
reasonable energy inputs
•Biology: realistic growth and
enzymatic rates, cell densities, PSE
• Interaction between variables
(e.g., cell density affects total
energy captured)
• Pilot and commercial capex and
operational cost for PBRs and open
ponds (with Tryg Lundquist, PhD at
Cal Poly)
Modelado Tecno-económico: valor de GPB
Secretion
• Enables continuous process through recycling of cells after secretion
• Most benefit through higher TAG Productivity
• Elimination of several costly process steps in TAG recovery
Oil metabolic engineering
• Improved redirection of light energy into TAGs
• Very high oil content levels not necessary with oil secretion
Other
• Identify additional bioengineering solutions
• Prioritize R&D projects based on synergies and on relative impact in cost
Validation
We can reach $50/bbl with realistic performance metrics for each of our
genetic engineering solutions
Process Flow Diagram Algae Biofuel Facility
                                Base Case                                                                  GPB Process
Make-up Water
                  Inoculation
                    Ponds                   Recirculation



                                                                 Algae Biomass
                                                                                                Blowdown
Make-up Water
                High Rate Algae                                                    Gravity
                                                Settling Basin
                    Ponds                                                         Thickener


                                                                                         Algae Biomass
                         CO2               Digestate

  Natural Gas
                                                 Anaerobic                       Solar Drying
                  Generator
                                    CH4           Digester                          Beds
                                     +
                                    CO2
                                                            Residuals                    Algae Biomass
                           Unrefined Oil
                                               Solvent Based
                                                                                 Flash Dryer
                                               Oil-Extraction




                Algae acts a catalyst that converts CO2 and sunlight into TAGs
                       efficiently, without being consumed in the process
  Work with Tryg Lundquist, Assistant Professor at Cal Poly
Modelado Tecno-económico
                                          Base Case    0%     50%     90%
                                    500
          Cost per barrel ($/bbl)
                                    400
                                    300
                                    200
                                    100
                                      0
                                           0 5 10 15 20 25 30 35 40 45 50 55 60
                                                      % Oil (w/w)
Early adoption of tecnoeconomic analysis provides:
     Quantification of value of biofuel solutions
     Guidance of R&D projects
     Validation

Presentación leandro vetcher

  • 1.
    Green Pacific Biologicals Junio 2012 Leandro Vetcher – CEO lvetcher@greenpacificbio.com @lvetcher Green Pacific Biologicals - Not for distribution
  • 2.
    GPB: Plataforma deingenieria genetica GPB Stable, stronger More efficient Access to the gene expression engineering entire cell other 0 50 transformants 50 % expressing % expressing transformants other GPB 0 Only company with rapid and stable eukaryotic algae nuclear genetic engineering Green Pacific Biologicals - PROPRIETARY and CONFIDENTIAL - Not for distribution 2
  • 3.
    GPB: Soluciones parabiocombustibles Increase carbon fixa on Implement efficient Maximize oil secre on carbon flux into oil Green Pacific Biologicals - PROPRIETARY and CONFIDENTIAL - Not for distribution 3
  • 4.
    Expresión génica nuclearmas fuerte GFP expression by Western blotting Accumulation of GFP by confocal microscopy GFP protein Cell extract Wild-type stds (ng) WT GPB GPB Strains 4
  • 5.
    Avg. fluorescence per cell (au) Avg. fluorescence per cell (au) Expresión génica nuclear estable 5
  • 6.
    Mas transformantes expresandotransgenes More rapid and efficient engineering Green Pacific Biologicals - PROPRIETARY and CONFIDENTIAL - Not for distribution 6
  • 7.
    RNAi funcional- ingenieríametabólica Genetic RNAi using GPB platform better than traditional mutant Starch lower Oil higher Growth normal 7
  • 8.
    Localización subcelular deproteína Membrane Lipid droplet localization localization Untransformed cell Membrane part+GFP Lipid droplet part+GFP 8
  • 9.
    Secreción de aceitesno-toxica  Non-toxic mechanism  Secreted LDs spontaneously separates, sidestepping harvesting/extraction  Works by interactions between proteins on LDs and cell membrane or other cellular components  Platform makes GPB uniquely positioned to implement oil secretion in algae 9
  • 10.
    Beneficios de secreciónde aceites no-toxica BASE CASE: GPB CASE: • Milk harvested by killing cow • Milk harvested through secretion and cow returned to pasture • Majority of energy  Making cow • Majority of energy (grass) goes into making milk • If cows are milked daily, there is no benefit to having cows that can store more than one day’s worth of milk MILK + MILK ?
  • 11.
    Modelado Tecno-económico Oil productivity model + operational & capital costs • Physics: energy balanced, reasonable energy inputs •Biology: realistic growth and enzymatic rates, cell densities, PSE • Interaction between variables (e.g., cell density affects total energy captured) • Pilot and commercial capex and operational cost for PBRs and open ponds (with Tryg Lundquist, PhD at Cal Poly)
  • 12.
    Modelado Tecno-económico: valorde GPB Secretion • Enables continuous process through recycling of cells after secretion • Most benefit through higher TAG Productivity • Elimination of several costly process steps in TAG recovery Oil metabolic engineering • Improved redirection of light energy into TAGs • Very high oil content levels not necessary with oil secretion Other • Identify additional bioengineering solutions • Prioritize R&D projects based on synergies and on relative impact in cost Validation We can reach $50/bbl with realistic performance metrics for each of our genetic engineering solutions
  • 13.
    Process Flow DiagramAlgae Biofuel Facility Base Case GPB Process Make-up Water Inoculation Ponds Recirculation Algae Biomass Blowdown Make-up Water High Rate Algae Gravity Settling Basin Ponds Thickener Algae Biomass CO2 Digestate Natural Gas Anaerobic Solar Drying Generator CH4 Digester Beds + CO2 Residuals Algae Biomass Unrefined Oil Solvent Based Flash Dryer Oil-Extraction Algae acts a catalyst that converts CO2 and sunlight into TAGs efficiently, without being consumed in the process Work with Tryg Lundquist, Assistant Professor at Cal Poly
  • 14.
    Modelado Tecno-económico Base Case 0% 50% 90% 500 Cost per barrel ($/bbl) 400 300 200 100 0 0 5 10 15 20 25 30 35 40 45 50 55 60 % Oil (w/w) Early adoption of tecnoeconomic analysis provides:  Quantification of value of biofuel solutions  Guidance of R&D projects  Validation

Editor's Notes

  • #3 Stable transgene expression Consistent transgene expression Much higher level of transgene expression More expressing transformants Access to all cellular compartmentsRNAi functional Strain health and stability #2 Algae has the ideal characteristics for an ideal renewable energy platform.satisfy increasing energy demands + mitigate effects of climate change+ protect increasingly scarce natural resource Minimize/avoid competition with arable land for agriculture High oil productivity per acre (10x-100X over plants) Use waste water and saline water, reducing competition for limited freshwater supplies Recycle carbon from CO2-rich flue emissions from power plants and other industrial emitters
  • #8 GPB rnai- gives benefits of starch knockout/mutant- more oil- without drawbacks on growth rate.
  • #11 A NOTE ON MILK STORAGE CAPACITY: There is a limit to how much milk a cow can produce in one day, since they can only process a fixed amount of grass (energetically limited) If cows are milked daily, there is no benefit to having cows that can store more than one day’s worth of milkBASE CASE: Once grown, cow produces milk which is harvested by killing cow Majority of available energy (grass) goes into making cow GPB CASE: Milk is harvested by secretion from cow (milking) and cows returned to pasture Roughly 100% available energy (grass) goes into making milk (once cow population is established)