nano-enabled materials for sustainable living, 2009 ... · pdms, pmma,) can be varied • protein...
TRANSCRIPT
Mankind Engineer’s Complexity from Simplicity!
Resistor
InductorCapacitor
Transistor
Diode
5 nm
Extracellular surface
Intracellular surface
H+
NH2
COOH
Membrane embedded region
Cyt c binding site
Sample Building Blocks of FunctionalitySample Building Blocks of Functionality
Valves & Active Filters
Motors Pumps
Transducers
Building with Molecular Machines
Cys-HisCys-HisCys-HisCys-HisCys-His
Cys-HisCys-HisCys-HisCys-His
BiotinCys-HisCys-His
Biotin
Streptavidin
Ni rodCGGSGGS-6xHisBiotinStreptavidinBiotinCys on γsubunitβ subunit-10xHisNi cap
Covalent
CovalentCovalent
Metal/Cluster
Metal/Cluster
BiospecificBiospecific
Intra-protein bonds
Working with single-molecule powered devices
500nm
Prerequisites for the Engineering of Functional Cytometobolic Systems
Coupled Protein Functionality
Environmental Stability
Path to Commercialization
Cell as a Model for the Synthetic Engineering Biological Systems
Membranes are the Key!• Precision placement of Molecules
• Compartmentalization
Elimination of Diffusion Effects & Enables Elimination of Diffusion Effects & Enables Locally High Concentrations forLocally High Concentrations for Improved Improved Reaction KineticsReaction Kinetics
MEMBRANES CONTROL MEMBRANES CONTROL ENERGY, MATTER, and INFORMATION FLOW!ENERGY, MATTER, and INFORMATION FLOW!
Engineering with Biomimetic Membranes
• Biomimetic membranes- At 10nm in thickness, these systems represent increased robustness over lipid systems (5nm).
• Cross-linkable methacrylate endgroups, UV irradiated free-radical polymerization.
• Block length, or block composition (i.e. PDMS, PMMA,) can be varied
• Protein viability established in block copolymer systems.
• Increased stability/lifetime (Lipid stability=days)
5nm
Lipid Bilayer
Hydrophilic layer, 2.5nm
Hydrophobic layer, 5nm
• Protein/Polymer Membrane Hybrid Nanodevice Schematics
• UV polymerizable endgroups enable increased stability
ABA Triblock Copolymer
Adapted from Tieleman et. al
PEtOz-PDMS-PEtOz
Mn = 7800, polydispersity index = 1.48
HO-(R)-PDMS-(R)-OH LiO-PDMS-OLi
(CH2)2 Cl(H2C)2Cl Si
(CH2)2 Cl(H2C)2Cl PDMS
N O
PDMS NOH
m
NOH
m
O O
+ n-BuLi
PDMS Nm-1
Nm-1
O O
N
O
I
O
NINaI, 100oC
KOH/MeOH
Poly(2-ethyloxazoline)-block-poly(dimethylsiloxane)-block-poly(2-ethyloxazoline)
TEM image
Artificial “Organelle” Provides Limitless Possible Technology Applications
• It is an orthodox membrane channel that only allows H2O to pass through it center.
• It excludes the passage of common contaminants including salts, urea, detergents, even protons.
• It is a highly stable protein that resists denaturing from acids, detergents, heat and voltage.
• It is easily harvested in milligram quantities from an engineered E. Coli strain.
Images sourced from http://ntmf.mf.wau.nl/aquaporin/background.htm
Conserved residues atprotein’s core create highlyselective water channel
Freeze Fracture TEM of Aquaporin Incorporated into Vesciles
MF cellulose membrane(Hydrophilic and High Biomaterial affinity)
Protein included polar lipids
AqpZ-polymersomes
Support Membrane
PrototypePrototypeAquaZ MembraneAquaZ Membrane
Small Plant
AquaZ Membrane Conventional RO
Memb. capacity; 600 psi/5.5 Mpa/40 bar 94.09 m3/d/m2 24859 gpd/m2 1.89 m3/d/m2 500 gpd/m2
Required membrane area 11 m2 114 ft2 526 m2 5,686 ft2
Unit electric power consumption 2.7 kWh/m3 2.7 kWh/m3 10 kWh/m3 10 kWh/m3
Large Plant
AquaZ Membrane Conventional RO
Memb. capacity; 600 psi/5.5 Mpa/40 bar 94.09 m3/d/m2 24859 gpd/m2 1.89 m3/d/m2 500 gpd/m2
Required membrane area 402 m2 4,328 ft2 20000 m2 215,199 ft2
Unit electric power consumption 1.36 kWh/m3 1.36 kWh/m3 5 kWh/m3 5 kWh/m3
AquaZ|Conventional RO Membrane AquaZ|Conventional RO Membrane ComparisonComparison
Universal Molecular Scale Energy Transduction
Vision
BR-ATPase Proteopolymersome
0 10 20 30 40 50 60-0.08
-0.06
-0.04
-0.02
0.00
0.02
Illuminated Dark
Time(minutes)
Del
ta p
H
0 10 20 30 40 50 60
0
1x104
2x104
3x104
4x104
5x104
AT
P pr
oduc
tion
(nm
ol A
TP/
mg
AT
Pase
)
Time(minutes)
ADP:50 μl (0.2 M) Pi:50 μl (1 M) Polymersome solution:355 μl
Nanoletters, 2005
Silica colloid
BR-ATP synthase integrated system
20 mM ADP0.4 M Pi
bR-ATPase Biogel Synthesis
SilicabR-ATP synthase liposome
Nature Materials, February 2005
Continuous production of ATP using microfluidic device
Solid-State ATP Generation System
Demonstration of Stable Bio-molecular System/Encapsulation in Solgel
Charging and Discharging Processes for the bR Proteoliposome Thin Gel
-0.05
-0.04
-0.03
-0.02
-0.01
0
0 20 40 60 80 100 120 140
chargingdischarging
chan
ge o
f pH
time (min)
(a)
7.3
7.35
7.4
7.45
7.5
7.55
7.6
7.65
7.7
pHtime (min)
0 25 0 25 0 30 0 25
Day 20 Day 21 Day 23 Day 27
(b)
Proteosomes retain recharge abilities for minimum of 27 days in proteogel conformation
Vision
Electrical Energy Electrical Energy from Lightfrom Light
Chemical Energy(ATP)
OpticalEnergy
ElectricalEnergy
Bacter
iorho
dops
in,
ATP S
ynth
ase
Bacteriorhodopsin,Cytochrome Oxidase
ATP Synthase,
Cytochrome Oxidase
Overview of Device Structure and Function
Depiction of Envisioned Device Packaging and Functionality
Description of COX Reversal/Electron Release Mechanism in Proteopolymersome
Sol-Gel Solid State System
hν
SolGel
Platinum
Nafion
SolGel
Gold
Overview of Device Structure and FunctionDescription of Active Element Reactions
A: Incident light causes conformational change in BR for proton pump across the membrane. This reaction convert the energy of photons into potential energy of protons.
C: COX is oxidased by Cytochrome C. Each Cytochrome C gain one electron so its charge goes from 3+ to 2+.
B: COX oxidases water using the energy of protons that come back into the vesicle throughout the protein.
D: Cytochrome C is oxidased in the anode passing out electrons.
COXred
COXox 4 Cytochrome Cox (3+)
4 Cytochrome Cred (2+)2 H2O
4H+ + O2
B C
4e
D
4e
E
E: Bilirubin Oxidase catalize the reduction of oxygen in the cathode.
2 H2O
4H+ + O2
A
BR
Light
BR convert light into a electrochemical potential
1 mol of photons190 kJ/mol100%
2 protons are pumped per photon
pH= 7.3ϕ = 0 mV
pH= 9.6ϕ = -137 mV
Free energy of each pumped proton = 26.75 kJ/mol
Energy stored = 53.5 kJ/mol28.2 %
A
Note: Considering that all the wave lengths are converted to wave lengths that can be absorbed by BR and that all the photons impact a protein.
BR
BR
+ + + + + + + + + +
- - - - - - - - - - - - - -
COX estimated efficiency = 65%Estimated Efficiency combining BR and COX = 18.3%
2 H2O 4H+ + O2 + 4e-
1 2 3 4 1 2 3 4
H+ e-
H+ e-
COXred
COXox 4 Cytochrome Cox (3+)
4 Cytochrome Cred (2+)2 H2O
4H+ + O2
B C
COX convert electrochemical potential into chemical energy
26.75 kJ/ mol of proton250.6 kJ/mol
pH= 7.3ϕ = 0 mV
pH= 9.6ϕ = -137 mV
+ + + + + + +
- - - - - - -
+ + + + + + + + + +
- - - - - - -- - - - - - -
BR COX
H+
35.9 kJ/ mol of electron
4 protons cross 4 electrons cross
Bio-solar Fuel Cell Function D: Cytochrome C is oxidized at the anode and releases its electrons.
4 Cytochrome Cox (3+)
4 Cytochrome Cred (2+)
4e
D
Electrode
Direct Current Measurement Using Fully-Assembled Vesicles Shows Light-Dependent Current Production
• Samples produced current based upon LIGHT/DARK environments
• Maximum duration switching ability= 2 hours
• Pt-Pt yeilds better response than Pt-Ag/AgCl- Increases electron harvesting/current production
•Maximum current production= 13.8 μA, or 0.273mA/cm2 with no applied voltage
Demonstrated Performance
Measured PerformanceDevice Dimensions ( cm 2) Min. 1x1
Currentmax (μA) 13.8
Voltagemax (V) 0.25
Power (μW) 3.45
Power Density (W/kg) 250Power/Area (W/m2)-Active 0.68
Conversion Efficiency (%)-Active, 350-750nmConversion Efficiency (%)- Active, 360-550nmConversion Efficiency (%)- Active, 500-550nmCurrent Density (mA/cm2)-ActiveActive= Area covered only by vesicles=10-17% conc.350-750nm=Entire Spectrum of Light used for calculation500-550nm= Spectrum covered only by green light360-550nm=Spectrum covered by "broadening" technologyLUX values used to calculate room light intensities fromCCD Arrays cameras and displays 2nd ed. by Gerald C. Holst. SPIE press ISBN 0-81294-2853-1 1998
1.2
3.0
6.30.273
Measurements using Fostec Light Source
Bioprocess Within a Bubble Architecture
Schematic of Bubble Structure
Cross-sectional view
Air
Soap Bubble
Soap Molecules
Polymer Vesicle
Water
Air Water
Soap Bubble and Polymer Vesicle Schematics
+ =
Schematic of Our Hybrid Bubble Device
Soap bubble
Dishwashing detergent
Functionalized Polymer Vesicles
Active membrane proteins incorporated into a polymer vesicle
Schematic of a Foam Structure
• When spherical bubbles come together, dry aqueous foam formation takes place forming polyhedra. • Plateau borders contain most of the aqueous solution.
Biochemical Synthesis within a Bubble Architecture
Hilgenfeldt, S., NAW 2002, 5/3, 224-230.
Synthesis within the Bubble Wall
ATP Synthesis within Bubble Architecture
0 10 20 30 40 50 60
0
400
800
1200
1600
AT
P pr
oduc
tion
(nm
ol/m
g A
TPa
se)
Time(minutes)
Buffer Bubble
• BR/BR-ATPase proteopolymersomes (without using ethanol)• Bubbles made using Tween
0 10 20 30 40 50 60-0.12
-0.08
-0.04
0.00
0.04
0.08
0.12
Buffer (pyranine inside vesicle) Bubble (pyranine outside vesicle) Bubble control (pyranine outside vesicle)
Del
ta p
H
Time(minutes)
ADP:30 μl (0.2 M) Pi:15 μl (1 M) Polymersome solution:300 μl
Carbon Fixation is the most sensitive and
slowest reaction in the Process.
Carbon Fixation is the most sensitive and
slowest reaction in the Process.
Polymersome ATP
Polymersome ATP
G3PG3P DHAPDHAP
Fructose-6-PhosphateFructose-6-Phosphate
Glucose-6-Phosphate Glucose
Glucose Production
Process Array Amount of Glucose (nmol)
Net Photoconversion Efficiency (%)
Chemical Conversion Efficiency from ATP (%)
All Separate Bulk 19 7.8 47.6
Foam 34 14.0 85.4
ATP Produced Separate
Bulk 18 7.4 45.1
Foam 35 14.5 88.4
Full System Bulk 21 8.7 53.0
Foam 38 15.7 95.7
Ethanol• Energy Density of 26.8 MJ/kg
• Water soluble
• Boiling point at 346 K
• Biomass produced at rate of 43 t/ha/a on a dry mass basis
• This yields available energy at maximal rate of 139 GJ/ha/a
• Water consumed from local sources at 800‐4200 L/L of ethanol produced
DMF from Simple Carbohydrate
• Energy Density of 37.5 MJ/kg
• Insoluble in Water
• Boiling point at 366 K
• Pure DMF produced at a rate of 69 t/ha/a
• This yields available energy at maximal rate of 2814 GJ/ha/a
• Under proper protocol nearly all water is reclaimed
Engineered “Metabolism” into MaterialsIncorporating Integrated Power, Amplified
Sensing, Biochemical Synthesis, Information Processing
Materials that See, Hear, Smell
The Montemagno Team
Join the Montemagno Team!
I’m looking for talented Graduate Students and Post Docs…contact me at:[email protected]