Nano-Enabled Materials for Sustainable Living, 2009 ... · PDMS, PMMA,) can be varied • Protein viability established in block copolymer systems. • Increased stability/lifetime

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]