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 Biofuels (Ethanol
Residues (biomass) & 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 Glycerol

Biodiesel Glycerin
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

6
6
Biodiesel Production Plants in 2006
U.S. Represents Two Billion Gallons of
Biodiesel Capacity

ISU

8
Biodiesel Production Today

Feed Oil Neutralization

Esterification Transesterification
Drying Water Wash

Water washing Biodiesel

Uses Non-renewable Catalyst and Neutralization
is necessary
(Sodium Methoxide or
Sodium Hydroxide)
Recovery Low quality High quality
Glycerin Glycerin

Catalyst contaminates
glycerin co-product

Methanol Toxic Waste

Residual water must be
distilled from methanol

Not all methanol recovered

U.S. Produces Biodiesel from Vegetable Oils

10
 Problems of
Free Fatty Acid (FFA)-Containing Feedstock

Animal Fat, Algae

Biodiesel
or Waste Oil
Base catalyst destroyed
by FFA’s (soap formation)

FFA Free Oil

Esterification Transesterification
by acid catalyst 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
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
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.
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.
19
19
Nanotechnology for
Extraction, Separation, and Fuel Production
Select
Strain

Nanofarming
20
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.

21
21
 Our Approach
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 H2O

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

R1 = 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.
 Mesoporous Mixed
Morphology Control
J. Am. Chem. Soc. 2003, 125, 4451 Metal Oxide
Chem. Mater. 2003, 15, 4247 Catalysts
New J. Chem. 2008, 32, 1311

Gate-keeping: Calcium Silicate

Selectivity Control Catalyst
Claisen
J. Am. Chem. Soc. 2004, 126, 1010
Rearrangement
Biodiesel Synthesis
Dalton Trans. 2009, 3237

Nitroaldol
Diels-Alder
Aldol
Michael Addition
J. Am. Chem. Soc. 2005, 127, 13305
Mesoporous Aluminum Silicate
Cooperative Catalysis with Single-Type Active Sites
Angew. Chem. Int. Ed. 2005, 44, 1826 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.

100.0
DTT
0.5 mm 90.0
No DTT

80.0

70.0

Fluorescent foci per cotyledon

UV light/GFP filter 60.0

50.0

40.0

30.0

20.0

10.0

0.0
MSN type Type IV Type IV Type III Type III

0.5 mm 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
C16:0
255.50 NL: 2.56E3
100
MCN_absorbed_TI_0802
90 C16:1 13151625#3-198 RT:
0.11-3.19 AV: 190 T:
80 ITMS - p MALDI Full ms
70 C16:3 [200.00-1000.00]

60
249.33 C18:1 *
291.33
50
C14:0 281.50
227.50 241.50
40 C18:3 C18:0 *
30 *
333.33
351.08

301.17
20
204.42
277.33
*
315.25
10 325.42 339.33 375.33
359.33 391.33
0
277.25 NL: 2.88E2
100
MCN_hex_neg_J19#2-
90 C18:3 120 RT: 0.02-2.01 AV:
119 T: ITMS - p MALDI
80 Full ms [150.00-1000.00]
Relative Abundance

Filter Paper 70
*
351.00
60
C16:3 *
291.25
50
(+MCN) 40
249.17
C16:0
30 C18:0
Hexane 20
301.08
*
333.25

Aqueous solution 10 204.25 233.25 263.25 *

317.17 339.25 367.17 377.25 391.33
0
277.25 NL: 7.04E2
100
MCN_H2O_neg_J22#2-
90 C18:3 82 RT: 0.02-1.32 AV: 81
(+MCN) 80
T: ITMS - p MALDI Full ms
[150.00-1000.00]
70 C16:3 *
291.25
*
351.08
01.08 60 249.25 C16:0
333.25 50 255.50
* Matrix
C22:0 40 C18:0
C20:0 325.42 339.3 30 301.08 *
333.25
* Polypropylene
311.33 20 227.42 241.42

10
213.25
263.25
311.33* 325.42 339.33

355.17 375.25 391.33

0
200 220 240 260 280 300 320 340 360 380 400
m/z
300 320 340
32
Selective Absorption of Fat Acid by MCN

Name Structure Absorptivity Dimension*

Myristic Acid L:18.624Å

Strong
(C14:0) W:3.101Å

Palmitoleic Acid L:18.899Å

Strong
(C16:1) W:8.0Å

Oleic Acid L:21.919Å

Strong
(C18:1) 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 L:21.172Å

Weak
(C16:0) W:3.101Å

Hexadecatrienoic L:19.407Å
Acid (C16:3)
Weak
W:5.079Å

Stearic Acid L:23.731Å

Weak
(C18:0) W:3.101Å

Gamma-linolenic
L:12.618Å
Acid Weak
(C18:3) 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 L:26.983Å

None
(C20:0) W:3.101Å

Behenic Acid L:29.026Å

(C22:0)
None
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)

100%
90%
sequestered by MSN from

80%
Percentage of FFAs

70%
hexanes

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
Biodiesel
Restaurant Oil
Base catalyst destroyed
by FFA’s (soap formation)

FFA Free Oil

Esterification Transesterification
by acid catalyst by base catalyst

Free Fatty Acid

We need a solid material that can catalyze both reactions!

 Our Bifunctional Solid Nanoporous
Catalytic System
Biodiesel
Free Fatty Acid

Biodiesel
Triglyceride

= 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
H2O

Hydrophobic Core
of Surfactant Micelle
 Synthesis of Mesoporous
Calcium Silicate Catalyst

Feedstocks
CTAB = N Br
n (Soybean Oil or
Animal Fat)

y = 6.50 for MCS-1 catalyst

= 3.25 for MCS-2 catalyst

Biodiesel
= 1.62 for MCS-3 catalyst
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.
 Recyclability of MCS-1
Soybean Oil: 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, SiO2 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 C3S:

2(CaO)3(SiO2) + 7H2O →
(CaO)3(SiO2)2•4(H2O) + 3Ca(OH)2
C-S-H CH
Hydration of C2S:

2(CaO)2(SiO2) + 5H2O →
(CaO)3(SiO2)2•4(H2O) + Ca(OH)2

60% of Portland cement is tricalcium silicate, Ca3SiO5, (C3S)

15% of Portland cement is β-dicalcium silicate, Ca2SiO4, (C2S)
Hydration of C3S and C2S 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, Ca5Si6O16(OH)2•7H2O

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)
0.279
(c) 0.182
(d) 0.166

(e)

Sample (a) is a hydrated C3S. Peaks associated with CH (Ca(OH)2) are marked with *.
Samples (b-e) obtained from decomposition of C3S in NH4NO3(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
Q4

MCM-41 Silica

Calcined MCS-3 Q3

Calcined MCS-2

Calcined MCS-1 Q2

As-synth MCS-1

West Central Sample Q1

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 29Si 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

Methoxide

Methanol Strip
Feed Tank Methanol HCl
Catalyst

Water Strip
Water Wash

Water Strip
Methanol Strip
Methyl Ester
Transesterification
Glycerol MeOH +
Gly + salts

Tech Glycerin Glycerin + Water + Finished

Glycerin Purification salts Gly + salts Biodiesel
Catilin Biodiesel Process

T300

Methanol Strip
Feed Tank Methanol
Catalyst

Methanol Strip
Methyl Ester
Transesterification Dry
Filter
Glycerol Wash

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