Chris Savile, Luan Nguyen, Oscar Alvizo, Svetlana Balatskaya, Michael Benoit, James Broering, Gerald Choi, Joshua

Geilhufe, Svetlana Gitin, John Grate, Jack Liang, Lisa Newman, Scott Novick, Jaime Parsons, James Riggins, Earl Solis, Sabrina Zimmerman, and Jim Lalonde
Codexis, Inc., 200 Penobscot Dr., Redwood City, CA 94063, USA Nexant Inc., San Francisco, CA jim.lalonde@codexis.com

Low-Cost CO2 Capture Using Enzyme Catalysis

Limitations of Current Solvent Capture Technology

High parasitic energy loss with MEA High heat of regeneration Predicted to nearly double cost of electricity from coal Solvents with lower energy loss tend to have very slow kinetics
MEA Accelerated-MDEA
Low due to bicarbonate Regeneration High due to formation formation; 30%+ lower Energy of carbamates total energy possible High; Low to Moderate Corrosiveness milli-inch / year = 136 milli-inch / year = 68 High; Low; Volatility Pamine= 6.7kPa @373K Pamine= 0.3 kPa @373 K Capture Rate Commercially Attractive ? Excellent No Excellent Yes

Approach: Acceleration with Carbonic Anhydrase

Directed Evolution to Create Stable CAs

5

CA Results to Date
4

Directed Evolution Strategy

Relative Activity (OD550)

Directed Evolution: tool to create an enzyme that is adapted to perform in harsh environments
a)
Obtain sequenceactivity data

4.2M MDEA, 24h 2.5 g/L CA

GS-02 Parent GSH1-137 Rd 1 hit GSH2-071 Rd 2 hit GSH3-095 Rd 3 hit GSH4-027 Rd 4 hit
Rd 5 hit GSH5-378 Rd 6 hit GSH6-046

3 2 1 0

35
-1

45

55

65 Challenge T (C)

75

85

95

b)

Statistical Analysis (ProSAR)

Compounded Fold Improvement

100000

PDB: 1SMJ - BM3 with palmitoyloleate5

CO2 + 2H2O

kCA

HCO3 + H3

+ O

10000

4.2 M MDEA, 87C; 1.5X 4.2 M MDEA, 85C; 2X 4.2 M MDEA, 70C; 3X 4.2 M MDEA, 65C; 2.5X 4 M MDEA, 50C; 9X

Carbonic anhydrase (CA) accelerates an otherwise negligible reaction. CA turnover rate up to 1 million CO2 [1] . molecules/s/s

c)

Beneficial Deleterious Neutral New backbone Existing diversity New diversity

1000

100

3 M MDEA, 42C (24h challenge); 6X

4.2 M MDEA >120C

d) e) f)

10

1 CA parent 1 2 3 Round of Evolution 4 5 6

New libraries

Enzyme Accelerated Process

PDB: 1BVY heme and reductase6

CA is a platform for any basic aqueous solution to accelerate the capture of CO2 as - . HCO 3 Process shifts toward bicarbonate formation, reducing overall heat of regeneration in the scrubber. Natural CAs tested not stable to T125C in solvents (10-50 wt% base). Evolved enzyme on trajectory towards surviving desorber process conditions.

CA + CO2 + H2O HCO3 + H

High Throughput Screen Challenge at high T and/or solvent H3O+ consumption via colorimetric dye Spectrophometric assay

Absorber column height (m)

CA screening: tiered approach

Challenge temperatures increased up to 87 C after 6 rounds of evolution.

250 200 150 100 50 0 1E+0 1E+1 1E+2 1E+3 1E+4 1E+5 Turnover Factor (TOF)

2 Medium Throughput Screen

Less solvent

Refine for process relevant criteria

Enzyme Enzyme

Increasingly harsh conditions (e.g., solvent, T, time)

Process emulation Low throughput

Less heat

Biocatalysis

Key Milestones and Deliverables

Year 1 Projected COE exceeded Year-1 milestone Developed biocatalyst stability exceeded (4X) Year-1 performance target Year 2 Develop biocatalyst with > 3 months stability Produce 100 kg enzyme for pilot plant runs Projected COE < 35% based on catalyst performance

Acknowledgement

Pilot Plant

This program is funded in part by the Advanced Research Projects Agency Energy (ARPA-E), U.S. Department of Energy, under Award Number DE-AR0000071
1. Khalifah, R.; Silverman, D. N., Carbonic Anhydrase Kinetics and Molecular Function, The Carbonic Anhydrase In Plenum Press: New York, 1991; pp 49-64. Acronyms COE: Cost of electricity, MDEA: N-methyldiethanolamine, MEA: monoethanolamine, ProSAR: Protein structure activity relationship.

The information, data, or work presented herein was funded in part by an agency of the United States Government. Neither the United States Government nor any agency thereof, nor any of their employees, makes any warranty, express or implied, or assumes any legal liability or responsibility for the accuracy, completeness, or usefulness of any information, apparatus, product, or process disclosed, or represents that its use would not infringe privately owned rights. Reference herein to any specific commercial product, process, or service by trade name, trademark, manufacturer, or otherwise does not necessarily constitute or imply its endorsement, recommendation, or favoring by the United States Government or any agency thereof. The views and opinions of authors expressed herein do not necessarily state or reflect those of the United States Government or any agency thereof.