1. Engineering Scalable Biocatalytic Processes
John M Woodley
Department of Chemical and Biochemical Engineering
Technical University of Denmark (DTU)
DK-2800 Lyngby
Denmark

2. Background

3. The Next Fifty Years 6-7 x 5-6 x 3.5 x 7 x Increase Increase
Global GDP growth (in constant dollars)
5-6 x
Production capacity for most commodities (steel, chemicals, lumber, etc.)
3.5 x
Energy demand
7 x
Electricity demand
Increase
Water demand
Increase
GHG emissions
Siirola (2012) Proc 11th Symp PSE (Ed Karimi and Srinivasan) 1

4. Features of BioProcesses
Effective conversion of sustainable (cheap, available, renewable) feedstocks
Operate under mild conditions
Renewable catalyst
Woodley et al (2013) Chem Eng Res Des 91, 2029

5. Types of Bioreaction Fermentation A+   A  B   B Microbial
biocatalysis
A+   A  B   B
A   A  B   B
A  B
Enzymatic biocatalysis

6. Types of Product from BioProcesses
Cost
High cost small molecules
e.g. pharmaceuticals (e.g. sitagliptin)
High cost large molecules
e.g. biopharmaceuticals (e.g. human growth hormone)
Molecular weight
Low cost small molecules
e.g. bulk chemicals (e.g. lactic acid)
Low cost large molecules
e.g. industrial enzymes (e.g. amylase)

7. Fermentation Process Structure
Separation 2
Fermenter
Separation 1 F P2 C

8. Challenges with Fermentation for Chemical Production
Diversion of carbon to make cells and by-products, limits yield
Conversion rate limited by growth, limited productivity
Product toxicity, limits concentration

9. Biocatalysis

10. Optimization of Biocatalyst Production and Conversion
Fermentation
Biocatalyst production
Biocatalysis
Conversion
Separate growth and conversion
Increased productivity by adding more biocatalyst

11. Increase Biocatalyst Yield
Fermentation
Biocatalysis
Recycle (whole-cell) biocatalyst

12. Features of Microbial Biocatalysis
Independent growth and conversion
Yield of reaction is set by the reaction (not growth)
Increased space-time yield (productivity) by addition of more biocatalyst
Potential to recycle biocatalyst
Marshall and Woodley (1995) Nature Biotech 13, 1072
Woodley (2006) Adv Appl Microbiol 60, 1

13. Reduce Cellular Constraints
Fermentation
Enzyme
isolation
Biocatalysis
Use (immobilized) enzyme to achieve high biocatalyst concentration

14. Features of Enzyme Catalysis
Exquisite selectivity
Operate under mild conditions
Enzyme from renewable resource
Ability to alter enzyme properties via protein engineering
Pollard and Woodley (2007) Trends Biotechnol 25, 66
Woodley (2008) Trends Biotechnol 26, 321
Sheldon and Woodley (2017) Chem Rev. DOI: /acs.chemrev.7b00203

15. Biocatalytic Process Structure
Separation 2 R
Reactor
Separation 1 P B
Lima-Ramos et al (2014) Green Proc Synth 3, 195

16. 1. Multi-step Catalysis

17. Biocatalysts for Multiple Steps
France et al (2017) ACS Catalysis 7, 710

18. 2. Chemo-enzymatic Catalysis

19. Combining Biocatalysis and Heterogeneous Inorganic Catalysis
Vennestrøm et al (2010) ChemCatChem 2, 249

20. Bio-petrochemicals

21. DTU - Novozymes Collaboration
Step 1
Step 2
Step 3
Bottles made of
Polyethylene terephthalate (PET)
Step 1: enzyme catalysed
Step 2 & 3 : cataylzed by inorganic catalysts
Boisen et al (2009) Chem Engng Res Dev 87, 1318
NOVOZYMES PRESENTATION
NOVOZYMES A/S 21

22. HMF as a Platform Chemical

23. DTU - Novozymes HMF Process
Solvent
Solvent separation
HMF recovery
G  F
F  HMF
Isomerization
Dehydration
HMF (crude)
Water, G,
“Other sugars”
Water, G,
“Other sugars”
Mixing
Glucose syrup
Abbreviations
F: Fructose
G: Glucose
HMF: Hydroxymethylfurfural

24. 3. Continuous Catalytic Processes

25. Michaelis-Menten Kinetics

26. Reactor Modelling
Kinetics for reactors obeying Michaelis-Menten kinetics:
Batch XS0 + Km(ln(1/(1 - X))) = kEt/V
Continuous plug-flow XS0 + Km(ln(1/(1 - X))) = kE/Q
Continuous flow stirred tank XS0 + Km(X/(1 - X)) = kE/Q

27. Biocatalytic Flow Reactors
Andrade et al (2014) Org Lett 16, 6092

28. Multiple Stirred Tanks

29. Agitated Cell Reactor
Toftgaard Pedersen et al Biotech Bioeng 114, 1222

30. Segmented Flow Reactor
Multi-phase options with organic solvent and aqueous phase

31. Tube-in-Tube Reactor
Teflon AF-2400 Tubing Inner Radius: 0.3 mm Tubing Thickness: 0.1 mm Reactor Length: 0.5 m Residence Times: 1-20 s Flow Rate: mL.min-1
Yang and Jensen (2013) Org Process Res Dev 17, 927

32. Mechanism and Rate Law

33. Tube-in-Tube Reactor (TiTR) for Enzyme Kinetic Analysis
Ringborg et al (2017) ChemCatChem 9, 3285

34. Comparison of TiTR with BSTR (1 atm)
Ringborg et al (2017) ChemCatChem 9, 3285

35. TiTR at Varying Pressure
Ringborg et al (2017) ChemCatChem 9, 3285

36. Role of Kinetics: Guiding Protein Engineering
Improved
process
Improved biocatalyst
Kinetic analysis
Woodley (2017) Phil Trans R Soc A. DOI: /rsta

37. Final Comments
Biocatalysis looks attractive for large scale processes due to:
excellent economic parameters (scalable)
ability to create new catalytic pathways (multi-step, chemo-enzymatic)
ability to operate processes continuously
At DTU we are always open to collaboration.

38. Special Thanks to Collaborators
Industries Universities
BASF (D) University of Manchester (UK)
Novozymes (DK) University of Stuttgart (UK)
DSM (NL) University of Groningen (NL)
c-LEcta (D) TU Graz (A)
Sigma Aldrich Chemie (CH) UAB (ES)
Novo Nordisk (DK)
InnoSyn (NL)
Lundbeck (DK)
Enzymicals (D)
Biosyntia (DK)

39. Contact Professor John M Woodley DTU Chemical Engineering jw@kt.dtu.dk