Nanofarming (with algae): Multifunctional Mesoporous ...

Nanofarming (with algae):
Multifunctional Mesoporous Nanoparticles for
Biofuel Production
Victor S.-Y. Lin
Department of Chemistry
Chemical & Biological Sciences Program
U. S. Department of Energy Ames Laboratory
Iowa State University
Ames, Iowa 50011-3111
U. S. A.

Global Warming?
Upsala Glacier in Argentina

The Debate is Over …
from Bob Willard
www.sustainabilityadvantage.com

From Biological Feedstocks to Energy
Different Resources
New Technologies
Different Applications
Food
Municipal Waste
Animal Feed
Short Rotation Trees
Electricity
Agricultural Crops,
Grasses, Algae, and
Residues (biomass)
Biofuels (Ethanol
& Biodiesel)
Hydrogen
Need new Refinery Methods for converting non-food resources into feedstocks.
Need new Catalysts that works at new interfaces.

Conversion of Oil to Biodiesel
Oil (Triglyceride)
Methyl Esters
Biodiesel
Glycerol
Glycerin

6
Advantages of Biodiesel
• Meets health effect testing (CAA – Clean Air Act)
• Lower emissions (VERY low/no N or S)
• High flash point (>300F)
• Biodegradable, essentially non-toxic
• Can be used in existing diesel engines
• Excellent lubrication properties
• Can use existing fuel distribution networks

Biodiesel Production Plants in 2006

U.S. Represents Two Billion Gallons of
Biodiesel Capacity
8
ISU

Biodiesel Production Today
Feed Oil
Drying
Esterification
Transesterification
Neutralization
Water Wash
Uses Non-renewable Catalyst
(Sodium Methoxide or
Sodium Hydroxide)
Water washing
and Neutralization
is necessary
Biodiesel
Recovery
Low quality
Glycerin
High quality
Glycerin
Catalyst contaminates
glycerin co-product
Methanol
Residual water must be
distilled from methanol
Toxic Waste
Not all methanol recovered

U.S. Produces Biodiesel from Vegetable Oils
10

Problems of
Free Fatty Acid (FFA)-Containing Feedstock
Animal Fat, Algae
or Waste Oil
Base catalyst destroyed
by FFA’s (soap formation)
Biodiesel
Esterification
by acid catalyst
FFA Free Oil
Transesterification
by base catalyst
Free Fatty Acid
We need a solid material that can catalyze both reactions!

Oil Feedstocks for Making Biodiesel
Table from: Y. Chisti, Biotechnol. Adv. 2007, 25, 294-
306.
• U.S. currently uses ~970 million acres for crops & grazing
• ~16.4 trillion acres of soybean would be needed for supplying ALL U.S.
transportation fuel.
• Only ~20 million acres of algae could supply the same amount of biodiesel, i.e., only
2.2 % of the existing U.S. cropping area would be needed

Oil Content of Microalgae
Microalgae
Table from: Y. Chisti, Biotechnol. Adv. 2007, 25, 294-306.

Green Microalgae: Botryococcus braunii (Bb)
14

Algae Continues to Garner R&D Interest and
Funding
15

Open Pond vs. Photobioreactor
Paddle wheels for mixing
General Atomics
Cyanotech Corp. in Hawaii
Solix Biofuels
16

From Algae to Biodiesel
Select
Strain
Picture modified from “The Fuel Cell”, Popular Science, 271(1), July 2007, Page 76-101.

Hydrocarbons from Green Microalgae
Ether Lipid & Triglyceride
Fatty Acids
Alkenes
Sterols
Botryococcus braunii (Bb)
(Race-A from U.K.)
Ref: (a) P. Metzger et al. Appl. Microbiol Biotechnol. 2005, 66, 486-496.
(b) N. O. Zhila et al. Russ. J. Plant Physiol. 2005, 52, 357-365.

19
Current Challenges
• Only short chain (< C20), non-branched hydrocarbons
are economical feedstocks for fuel production.
• Current oil extraction methods cause fatal damages to the
cell growth of most, if not all, algae.
• Lack of efficient and economical refinery methods for
the isolation of the suitable fuel feedstocks from the
“alphabet soup” of hydrocarbons from algae.
• Current commercial catalysts (e.g. NaOMe) for biodiesel
production cannot handle the impurities, such as free
fatty acids and lipid phosphoric acids, of algae oil.

Nanotechnology for
Extraction, Separation, and Fuel Production
20
Select
Strain
Nanofarming

21
Our Approach to Overcome these Challenges
• Develop non-invasive oil extraction methods with
high recyclability and regrowthability of algae.
• Design new sequestration method for selective
isolation of the suitable fuel feedstocks from the
hydrocarbons of algae.
• Construct solid catalysts for efficient production of
biodiesel from algae oils with fatty acids.

Our Approach:
Introduce functional groups that are electrostatically or hydrophobically attractive to
the ammonium surfactant head groups and able to compete with silicate anions.
Aqueous Phase
H 2 O
H 2
Hydrophobic Core
of Surfactant Micelle
Lin, V. S.-Y.; Lai, C.-Y.; Huang, J.; Song, S.-A.; Xu, S. J. Am. Chem. Soc. 2001, 123, 11510-11511.
Huh, S.; Wiench, J. W.; Yoo, J.-C.; Pruski, M., Lin, V. S.-Y. * Chem. Mater. 2003, 15, 4247-4256.
Huh, S.; Wiench, J. W.; Trewyn, B. G.; Pruski, M.; Lin, V. S.-Y. * Chem. Comm. 2003, (18), 2364 - 2365

Interfacial Hydrophobic and Electrostatic Interaction
between Organosilicates and Micellar Surfactants (CTAB)
R = Hydrophobic Functional Groups
R 1 = Hydrophilic Functional Groups
Lin, V. S.-Y.; Lai, C.-Y.; Huang, J.; Song, S.-A.; Xu, S. J. Am. Chem. Soc. 2001, 123, 11510-11511.
Huh, S.; Wiench, J. W.; Yoo, J.-C.; Pruski, M., Lin, V. S.-Y. * Chem. Mater. 2003, 15, 4247-4256.
Huh, S.; Wiench, J. W.; Trewyn, B. G.; Pruski, M.; Lin, V. S.-Y. * Chem. Comm. 2003, (18), 2364 - 2365

FE-SEM of Organically Functionalized Mesoporous
Silica Materials with Different Particle Morphologies
Field Emission Scanning Electron Micrographs:
APTMS AAPTMS AEPTMS UDPTMS
CPTES ICPTES ATMS Pure MCM-41
Huh, S.; Wiench, J. W.; Trewyn, B. G.; Pruski, M.; Lin, V. S.-Y. Chem. Comm. 2003, 2364-2365.
Huh, S.; Wiench, J. W.; Yoo, J.-C.; Pruski, M., Lin, V. S.-Y. Chem. Mater. 2003, 15, 4247-4256.

Organically Functionalized Mesoporous Silica Nanosphere
• Transmission Electron Micrograph:
Lin, V. S.-Y.; Lai, C.-Y.; Huang, J.; Song, S.-A.; Xu, S. J. Am. Chem. Soc. 2001, 123, 11510-11511.
Lai, C.-Y.; Trewyn, B.G.; Jeftinija, D.M.; Jeftinija, K.; Xu, S.; Jeftinija, S.; Lin, V.S.-Y. J. Am. Chem. Soc., 2003, 125, 4451-4459.

Morphology Control
J. Am. Chem. Soc. 2003, 125, 4451
Chem. Mater. 2003, 15, 4247
Mesoporous Mixed
Metal Oxide
Catalysts
New J. Chem. 2008, 32, 1311
Gate-keeping:
Selectivity Control
J. Am. Chem. Soc. 2004, 126, 1010
Nitroaldol
Diels-Alder
Aldol
Michael Addition
J. Am. Chem. Soc. 2005, 127, 13305
Cooperative Catalysis
Angew. Chem. Int. Ed. 2005, 44, 1826
Calcium Silicate
Catalyst
Claisen
Rearrangement
Biodiesel Synthesis
Dalton Trans. 2009, 3237
Mesoporous Aluminum Silicate
with Single-Type Active Sites
J. Phys. Chem. C 2007, 111, 1480

Capped Mesoporous Silica Nanoparticles (MSN) as a
Multifunctional Cell Membrane Permeable Delivery Carrier
Intracellular Controlled Release
Cell Membrane
Enzyme or
Chemicals
Gene Expression
Plasmid DNA
Our recent review articles:
Giri, S.; Trewyn, B. G.; Lin, V. S.-Y.* Nanomedicine. 2007, 2(1), 99-111.
Trewyn, B. G.; Slowing, I. I.; Giri, S.; Lin, V. S.-Y.* Chem. Commun. (Feature Article), 2007, 3236-3245.
Slowing, I. I.; Trewyn, B. G.; Giri, S.; Lin, V. S.-Y.* Adv. Funct. Mater. (Feature Article), 2007, 17, 1225-1236.

Animal versus Plant Cells

Gold Nanoparticle Capped MSN as a Multifunctional
Cell Wall Permeable Delivery Carrier for Plants
MSN
β-Estradiol
DTT
Gene Expression
Plasmid DNA
Torney, F.; Trewyn, B. G.; Lin, V. S.-Y.,* Wang, K.* Nature Nanotech. 2007, 2, 295-300.

GFP Expression in Tobacco by Gold Nanoparticle
Capped MSN-mediated Delivery of β-Estradiol
Bright field
Non-transgenic Tobacco in DTT medium
bombarded by gfp DNA-coated Type III
(fluorescein loaded) or Type IV (β-estradiol
loaded) MSNs.
0.5 mm
100.0
90.0
80.0
DTT
No DTT
UV light/GFP filter
Fluorescent foci per cotyledon
70.0
60.0
50.0
40.0
30.0
20.0
10.0
0.5 mm
0.0
MSN type Type IV Type IV
Type III
Type III
MSN content β-Estradiol β-Estradiol FITC FITC
Event
B G B G
Torney, F.; Trewyn, B. G.; Lin, V. S.-Y.,* Wang, K.* Nature Nanotech. 2007, 2, 295-300.

Mesoporous Carbon Nanosphere with Algae
• MCN has very high affinity toward Algae:
TEM of MCN:
Green Microalgae
(Botryococcus braunii; Bb)
Bb with MCN
Kim, T.-W.; Chung, P.-W.; Slowing, I. I.; Tsunoda, M.; Yeung, E. S.; Lin, V. S.-Y.* Nano Lett, 2008, 8, 3724-3727.

MALDI Mass Spectroscopy Profile
01.08
C20:0
311.33
Filter Paper
(+MCN)
Hexane
Aqueous solution
(+MCN)
333.25
C22:0
325.42 339.3
300 320 340
Relative Abundance
100
90
80
70
60
50
40
30
20
10
0
100
90
80
70
60
50
40
30
20
10
0
100
90
80
70
60
50
40
204.42
204.25
C14:0
C16:1
C16:3
249.33
227.50 241.50
C16:3
249.17
255.50
233.25 263.25
C16:3
249.25
C16:0
255.50
277.33
291.33
281.50
C18:3 C18:0
C18:3
C16:0
C18:3
C16:0
C18:1
277.25
277.25
291.25
C18:0
*
*
*
291.25
C18:0
301.17
301.08
333.33
351.08
315.25
325.42 339.33 359.33 375.33
391.33
333.25
351.00
317.17 339.25 367.17 377.25
351.08
391.33
30
301.08
333.25
20
227.42 241.42 263.25 325.42 339.33
213.25 311.33*
10
355.17 375.25 391.33
0
200 220 240 260 280 300 320 340 360 380 400
m/z
*
*
*
*
*
*
*
*
NL: 2.56E3
MCN_absorbed_TI_0802
13151625#3-198 RT:
0.11-3.19 AV: 190 T:
ITMS - p MALDI Full ms
[200.00-1000.00]
NL: 2.88E2
MCN_hex_neg_J19#2-
120 RT: 0.02-2.01 AV:
119 T: ITMS - p MALDI
Full ms [150.00-1000.00]
NL: 7.04E2
MCN_H2O_neg_J22#2-
82 RT: 0.02-1.32 AV: 81
T: ITMS - p MALDI Full ms
[150.00-1000.00]
* Matrix
* Polypropylene
32

Selective Absorption of Fat Acid by MCN
Name Structure Absorptivity Dimension*
Myristic Acid
(C14:0)
Strong
L:18.624Å
W:3.101Å
Palmitoleic Acid
(C16:1)
Strong
L:18.899Å
W:8.0Å
Oleic Acid
(C18:1)
Strong
L:21.919Å
W:10.0Å
* L and W represent length and width of molecule, respectively.

Selective Absorption of Fat Acid by MCN
Name Structure Absorptivity Dimension*
Palmitic Acid
(C16:0)
Weak
L:21.172Å
W:3.101Å
Hexadecatrienoic
Acid (C16:3)
Weak
L:19.407Å
W:5.079Å
Stearic Acid
(C18:0)
Weak
L:23.731Å
W:3.101Å
Gamma-linolenic
Acid
(C18:3)
Weak
L:12.618Å
W:12.521Å
* L and W represent length and width of molecule, respectively.

Selective Absorption of Fat Acid by MCN
Name Structure Absorptivity Dimension*
Arachidic Acid
(C20:0)
None
L:26.983Å
W:3.101Å
Behenic Acid
(C22:0)
None
L:29.026Å
W:3.101Å
* L and W represent length and width of molecule, respectively.

MSN will Sequester Highly Valued Products
(carotenoids, vitamins, polysaccharides, and essential fatty acids)
Percentage of FFAs
sequestered by MSN from
hexanes
100%
90%
80%
70%
60%
50%
40%
30%
20%
10%
0%
C16:1 C16 C18:2 C18:1 C18 C20 C22
Procedure: A hexane solution of 7 common algal
FFAs was passed through the column shown on
right by gravimetric force. Graph above illustrates
the percentage of FFAs sequestered by MSN from
the hexane solution.
MSN
Sea Sand

Problems of
Free Fatty Acid (FFA)-Containing Feedstock
Animal Fat or
Restaurant Oil
Base catalyst destroyed
by FFA’s (soap formation)
Biodiesel
Esterification
by acid catalyst
FFA Free Oil
Transesterification
by base catalyst
Free Fatty Acid
We need a solid material that can catalyze both reactions!

Free Fatty Acid
Our Bifunctional Solid Nanoporous
Catalytic System
Biodiesel
Triglyceride
Biodiesel
= FFA Catalyst
= Oil Catalyst
Glycerol
Victor Lin Group, Iowa State University, U. S. A.

Our Approach: Modified Co-condensation
Can we introduce “non-siliceous” species, such as metal oxides and metal
complexes in the surfactant-templated co-condensation reaction?
Aqueous Phase
H 2 O
Hydrophobic Core
of Surfactant Micelle

Synthesis of Mesoporous
Calcium Silicate Catalyst
CTAB = N Br
n
Feedstocks
(Soybean Oil or
Animal Fat)
y = 6.50 for MCS-1 catalyst
= 3.25 for MCS-2 catalyst
= 1.62 for MCS-3 catalyst
Biodiesel
MCS
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Catalyst Performance for Soybean Oil
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Soybean Oil:
Recyclability of MCS-1
Poultry Fat:
Recyclability for Soybean Oil: 20 recycles without
decrease in reactivity was observed.
Recyclability for Poultry Fat: 8 recycles with small
decrease in reactivity was observed.
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Are MCS Catalysts Basic or Acidic?
Feedstocks
(Soybean Oil or
Animal Fat)
Biodiesel
CaO is a basic catalyst for Transesterification of Oil
However, SiO 2 is not acidic enough to catalyze Esterification of FFA!
How did the Esterification of Free Fatty Acids happen?
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Powder X-Ray Diffraction
Commercial Calcium Oxide:
As-synthesized MCS-1
Catalyst:
0.304
0.279
0.182
0.166
There is no crystalline CaO domain in MCS-1 material!
Are MCS materials some kind of calcium silicate mixed oxides?
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Calcium Silicates (Cement)
Portland Cement:
Hydration of C 3 S:
2(CaO) 3 (SiO 2 ) + 7H 2 O →
(CaO) 3 (SiO 2 ) 2 •4(H 2 O) + 3Ca(OH) 2
C-S-H
Hydration of C 2 S:
CH
2(CaO) 2 (SiO 2 ) + 5H 2 O →
(CaO) 3 (SiO 2 ) 2 •4(H 2 O) + Ca(OH) 2
60% of Portland cement is tricalcium silicate, Ca 3 SiO 5 , (C 3 S)
15% of Portland cement is β-dicalcium silicate, Ca 2 SiO 4 , (C 2 S)
Hydration of C 3 S and C 2 S yield calcium silicate hydrate (C-S-H)

Can it be a C-S-H?
C-S-H(I): Structure similar to the 1.4 nm Tobermorite, Ca 5 Si 6 O 16 (OH) 2 •7H 2 O
Along [001] Along [210] Along [010]
Calcium atom
Silicate tetrahedra

Powder X-Ray Diffraction
C-S-H(I) gels: a
As-synthesized MCS-1
Catalyst:
0.304
(a)
(b)
(c)
(d)
(e)
0.279
0.182
0.166
Sample (a) is a hydrated C 3 S. Peaks associated with CH (Ca(OH) 2 ) are marked with *.
Samples (b-e) obtained from decomposition of C 3 S in NH 4 NO 3 (aq).
a
Jennings, M. M. et al.; Cement and Concrete Research, 2004, 34, 1499-1519.

Powder X-Ray Diffraction
Calcined MCS Catalysts:
As-synthesized MCS-1 Catalyst:
0.304
MCS-3
0.279
0.182
0.304
MCS-2
0.166
0.182
MCS-1
Peaks with d-spacing 0.304 and 0.182 are preserved after calcination.
This indicated that the two-dimensional layers of calcium oxide and silicate
chains did not get destroyed during the calcination.
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Solid State NMR
Q 4
MCM-41 Silica
Calcined MCS-3
Q 3
Calcined MCS-2
Calcined MCS-1
Q 2
As-synth MCS-1
West Central Sample
Q 1
Portland Cement
X = Ca or H
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Possible Structure Transformation
From Q2 to Q3 silicates:
How does the structural change impact the catalytic reactivity?
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Structure-based Reactivity
Ca/Si Ratios of MCS Catalysts from 29 Si Spin Counting :
MCS-1 MCS-2 MCS-3
Ca/Si 1.7 1.0 0.2
• Observations:
– The reactivity against high FFA-containing oils is
significantly enhanced as the ratio of Ca/Si increases.
– Surface chemisorption analysis confirmed the presence of
Lewis acidic sites.
– The surface concentration of Lewis acidic sites increases as
the Ca/Si ratio becomes higher.
V. S.-Y. Lin, J. A. Nieweg, J. G. Verkade, C. R. V. Reddy, C. Kern, U.S. Patent Application (US 2008/0021232 A1), Jan. 24, 2008.

Catilin Inc.
A Joint Venture Company of Biodiesel Catalyst Technology
between Victor Lin, Iowa State University, and Mohr Davidow
Ventures (Menlo Park, CA)
http://www.catilin.com

Traditional Biodiesel Process
Esterification
Water
Feed Tank
Transesterification
Methanol
Methoxide
Catalyst
Methyl Ester
Glycerol
HCl
MeOH +
Gly + salts
Methanol Strip
Methanol Strip
Water Wash
Water Strip
Water Strip
Tech
Glycerin
Glycerin
Purification
Glycerin +
salts
Water +
Gly + salts
Finished
Biodiesel

Catilin Biodiesel Process
Feed Tank
Transesterification
Methanol
T300
Catalyst
Methyl Ester
Glycerol
Methanol Strip
Methanol Strip
Filter
Dry
Wash
Tech
Glycerin
Finished
Biodiesel

Exceptional Value Proposition
• Saves >10¢ per gallon
• Lowers capital costs by 50%
• Recyclable and non-toxic
• Produces cleaner Biodiesel &
Glycerol
• Can be used in existing
facilities with minimal
modifications
• Can be used with multiple
feedstocks
55

CATILIN: Commercializing ISU Technologies
Team Catilin in front of Biodiesel Pilot Plant
Catilin’s T-300 Biodiesel Catalyst
Catilin’s Research Labs on ISU Campus
Catilin’s Pilot Plant at BECON, Navada, IA

Catilin’s Continuous Flow Biodiesel Pilot Plant

• ISU Department of Chemistry:
Lin Research Group:
– Former Members:
Dr. Cheng-Yu Lai
Dr. Supratim Giri
Dr. Carla Wilkinson
Acknowledgement
Dr. Dana Radu
Dr. Jennifer Nieweg
Dr. Yang Cai
– Current Postdoctoral Researchers:
Dr. Igor Slowing Dr. Brian G. Trewyn
Dr. Hung-Ting Chen
Dr. Tae-Wan Kim
Dr. Tse-Min Hsin
– Current Ph.D. Graduate Students:
Mr. Juan Vivero-Escoto Mr. Cedric Chung
Mr. Robert Roggers
Ms. Wei Huang
Mr. Chih-Hsiang Tsai
Ms. Yannan Zhao
Mr. Justin Valenstein
Mr. Kapil Kandel
Mr. Tianfu Wang
• U.S. DOE, Ames Laboratory
Mr. Yulin Huang
Ms. Chorthip Peeraphatdit
Ms. I-Ju Fang
Ms. Enro Guo
Mr. Nikola Knezevic
Mr. Xiaoxing Sun
Dr. Marek Pruski’s Research Group
• $$$ Funding $$$:
– U.S. DOE, Office of Basic Energy
Sciences (AL-03-380-011)
– U.S. DOE, EERE (DE-FG26-
0NT08854 )
– NSF: CAREER Award (CHE-
0239570) and (CHE-0809521)
– USDA NRI-Biorenewable
– Pioneer Hi-Bred International, Inc