Electrocatalyst Development for Hydrogen Production and Utilization

Daniel SobczynskiSURE ProgramMaroney Group

Chrisjoe A. Joseph, Project LeaderJulius Campecino, Graduate Student Mentor

OverviewWhy Hydrogen?Hydrogenase SpecificsIsolation of the Ni-Fe HydrogenaseHydrogen Production Hydrogen UtilizationBenefits of the ProjectFuture Studies

Why Hydrogen? Can be used for

electrochemical cellsCan be formed by

biological catalystsClean emissions

Image credit: www.autopten.com

Hydrogenase Specifics

H2 2H+ 2e-+

H2 D2O+ HD + HDO

o-H2 p-H2

Image credit: Prof. Michael Maroney

Growing and Harvesting the cells Thiocapsa

roseopersicinaPhototropicMarine bacteriaWithstands oxygen

and non-oxygen environments

Centrifuge

Cell paste, stored at -20˚C

Image credit: Julius Campecino

Hydrogenase Purification

supernatant

DEAE: Anion-exchange resin

filtrate DEAE with bound proteins

450mM NaCl + 20mM Tris, pH 7.5

DEAE

clean DEAE

filtrate

filtrate butyl sepharose column*

butyl sepharose column1mM TRIS buffer, pH 7.5

Q sepharose column20mM TRIS buffer, pH 7.5

Q sepharose column50mM MES buffer, pH 5.5

Q sepharose columnTRIS buffer, pH 7.5

pure hydrogenase

Dissolve 15g acetone powder in water

100 grams of ammonium sulfate,butyl sepharose resin

300g cell paste

acetone powder

pellet

1M NaCl + 20mM Tris, pH 7.5

native gel electrophoresis(9% gel)

Electrocatalyst Design

Images credit: Prof. Dhandapani Venkataraman

Production of Hydrogen

O2Ti O2TiO2Ti

Image credit: Hydrogenase picture: Prof. Dhandapani Venkataraman

Hydrogen Utilization

Image credit: Hydrogenase picture: Prof. Dhandapani Venkataraman

Additional Modification Site

Proximal Medial Distal

Images credit: Prof. Dhandapani Venkataraman

Benefits of the ProjectVersatility of the

projectAlternative to

hydrogen storage issue

Water as output, and possibly input too

Image credit:Water drop: http://www.biofeedbackcalifornia.org

Future StudiesOptimizing hydrogen

production and utilization

Finding a way to better produce hydrogenase

Integrating the project goals into applied fields of science

Butyl Seph, Tris ChromatogramMax UV: 2200 mAU

%B concentration: 0 – 100 %

Conductivity range: 100 – 0 mS/cm

Fractions collected: All

H2ase Butyl 2 062409:10_UV H2ase Butyl 2 062409:10_Cond H2ase Butyl 2 062409:10_Conc H2ase Butyl 2 062409:10_Fractions

-500

0

500

1000

1500

2000

mAU

50 100 150 200 250 300 350 400 mlX1 X2 X3 X4 X5 X6 X7 X8

Q-Seph, Tris Chromatogram H2ase Long QSeph A load002:10_UV H2ase Long QSeph A load002:10_Cond H2ase Long QSeph A load002:10_Conc H2ase Long QSeph A load002:10_Fractions

500

1000

1500

2000

2500

mAU

220.0 230.0 240.0 250.0 minX1 X2 X3 X4

Max UV: 2600 mAU

%B concentration: 60 %

Conductivity range: 25 – 50 mS/cm

Fractions collected: X2 , X3

Q-Seph, MES Chromatogram H2ase MES QSephB load B 071209C001:10_UV H2ase MES QSephB load B 071209C001:10_Cond H2ase MES QSephB load B 071209C001:10_Conc H2ase MES QSephB load B 071209C001:10_Fractions

0

50

100

150

200

250

mAU

160 180 200 220 240 mlWaste X3 X4 Waste

Max UV: 230 mAU

%B concentration: 17.5 – 60 %

Conductivity range: 17.5 – 50 mS/cm

Fractions collected: X3 , X4

Pure Hydrogenase, EPR Results

Buffer Calculations450 mM NaCl, 20mM Tris buffer, pH = 7.5

For 1L of buffer:

(Mol. Wt.) x (# of Mol) = 58.443g/mol x .45 mol = 26.29935 g NaCl

M1 * V1 = M2 * V2 Solve for V2

20mM * 1L = 1M * V2

V 2=20 mL of 1M Tris