Available online at www.sciencedirect.
com
Aquatic phototrophs: efficient alternatives to
land-based crops for biofuels
G Charles Dismukes1, Damian Carrieri1, Nicholas Bennette1,
Gennady M Ananyev1 and Matthew C Posewitz2
To mitigate some of the potentially deleterious environmental
and agricultural consequences associated with current land-
based-biofuel feedstocks, we propose the use of biofuels
derived from aquatic microbial oxygenic photoautotrophs
(AMOPs), more commonly known as cyanobacteria, algae, and
diatoms. Herein we review their demonstrated productivity in
mass culturing and aspects of their physiology that are
particularly attractive for integration into renewable biofuel
applications. Compared with terrestrial crops, AMOPs are
inherently more efficient solar collectors, use less or no land,
can be converted to liquid fuels using simpler technologies than
cellulose, and offer secondary uses that fossil fuels do not
provide. AMOPs pose a new set of technological challenges if
they are to contribute as biofuel feedstocks.
Address
1 cannot be met solely by terrestrial crops. Multiple com-
Department of Chemistry and Princeton Environmental Institute,
Princeton University, Princeton, NJ 08544, USA
2
Department of Chemistry and Geochemistry, Colorado School of
Mines, Golden, CO 80401, USA
Corresponding author: Dismukes, G Charles (dismukes@Princeton.EDU)
Current Opinion in Biotechnology 2008, 19:235240
This review comes from a themed issue on
Energy Biotechnology
Edited by Lee R. Lynd
Available online 6th June 2008
0958-1669/$ see front matter
# 2008 Elsevier Ltd. All rights reserved.
DOI 10.1016/j.copbio.2008.05.007
Introduction
The scientific community has recently focused consider-
able attention on developing viable renewable biofuels as
leading alternatives to fossil energy in order to address the
issue of global warming. However, recent headlines have
generally condemned biofuels (e.g. [1]) owing to their
potential to drive up food prices and exacerbate CO2
release through the forced clearing of natural ecosystems,
which are as effective or more efficient in capturing CO2
[2,3]. The topic is emotionally charged, with divisive
positions held by laypeople and scientists alike, making
policy choices controversial and their economic and
environmental outcomes potentially devastating. In order
to minimize the potentially deleterious environmental
and agricultural consequences associated with current
www.sciencedirect.com
land-based-biofuel feedstocks, it is possible to include
biofuels derived from aquatic microbial oxygenic photo-
autotrophs (AMOPs), more commonly known as cyano-
bacteria, algae, and diatoms, into the bioenergy portfolio.
The potential of AMOPs as high-yield sources for lipids
(2050% dry wt) and fermentable biomass (starch and
glycogen, 2050% dry wt) was documented in research
conducted by the National Renewable Energy Laboratory
(NREL) and its contractors within the Aquatic Species
Program (NRELASP) during the 1980s1990s [4].
Recent workshops including the NRELAFOSR work-
shop on Algal Oil for Jet Fuel Production held in February
2008 concluded that ethanol cannot substitute for energy-
dense diesel for aviation fuels, and the latter demand
mercial ventures involving collaborations between large
oil companies and research institutions have recently
emerged to produce biodiesel from AMOPs, including
Chevron and NREL, Shell Oil and researchers at the
University of Hawaii and other institutions, and British
Petroleum and Arizona State University.
Herein we summarize the demonstrated productivity of
mass-cultured AMOPs and examine both their potential
and their limitations for the production of biomass and
biofuel precursors. We consider options for their large-
scale cultivation in marine or non-arable lands that would
augment or even enable a transition away from conven-
tional biomass grown on farmlands.
Current commercial and laboratory
productivity of various AMOPs
There is extensive worldwide experience in commercial
scale growth of food-grade AMOPS [5,6]. This experience
is limited to the 5 ha scale using open ponds. Growth data
from NREL and other reliable sources are summarized in
Table 1 and S1 listing the solar biomass productivity (dry
metric tons/ha  yr) for selected AMOPS grown year-rou-
nd at solar fluxes common to the American southwest, in
stirred open pond reactors at (suboptimal) ambient tem-
peratures, and with or without CO2 supplementation.
Compared with genetic hybrid strains of corn grain, the
annual biomass yield from native strains of AMOPs is 5.4
10 fold greater, while the comparable gain for switchgrass is
2.5 (fertilized arable land) to 10 fold (fertilized mixed
prairie grasses, footnote a, Table 1). We have converted
these biomass yields to raw energy content (GJ/ha  yr;
footnote f, Table 1). The range for AMOPs is 7001550,
Current Opinion in Biotechnology 2008, 19:235240
236 Energy Biotechnology

Table 1

Biomass and energy productivities of land-based plants and mass-cultured aquatic microbial oxygenic phototrophs (algae and
cyanobacteria)

Outdoor, [1_TD$IF]solar demonstrated values Corn [2_TD$IF]grain Sugarcane Switchgrass Rape Tetraselmis Arthrospira
(except in parentheses) and mixed [3_TD$IF]seeds suecica (Spirulina)
prairie grasses species
Productivity (dry metric tons/ha [4_TD$IF] yr) 7 [25] 7387 [25] 3.615[5_TD$IF]a [26,27] 2.7 [28] 3869[6_TD$IF]b [4] 27c, 6070 d
Productivity raw energy (GJ/ha  yr) e 120 [25] 12301460 [25] 61255 [27] 73 [28] 7001550[7_TD$IF]f 550, 12301435
Components [25,29] [25] [25,30] [31] [4] [6,8,32]
[8_TD$IF]Nonrecalcitrant [9_TD$IF]carbohydrates (%) 70 30 4.511.5 (11)g(47) h 15g(50) h
Lipids (%) 4.56 13 11.6 42 (23)[10_TD$IF]g(15) h 5g(13) h
Protein (%) 612 (68)g(28) h 72g(27) h
Water usage (L/dry kg) 565 [25] 89118 [33] 50 [34] 3390 [35] 310570 [4]
Water [1_TD$IF]usage per energy (L/MJ) e 33 57 3 200 1834
a
Mixed prairie grass data reported here involve field burning of the annual crop rather than harvesting, thereby enhancing productivity by self-
fertilization at the cost of eliminating biomass utilization (zero yield).
[12_TD$IF]b
Lower number demonstrated full year, upper number demonstrated in summer months in New Mexico. Monoraphidium minutum (MONOR2) was
also used for growth experiments.
[13_TD$IF]c
Food grade in Mexico [6] or grown on seawater and urea rather than standard media in Italy [36].
[14_TD$IF]d
With heated ponds, control of photoinhibition, and proper harvest timings [37].
[15_TD$IF]e
Assuming heat of combustion, theoretical maximum energy content.
[16_TD$IF]f
Assuming heat of combustion energy similar to A. maxima 2 kJ/g (A. maxima combustion energy = 20.5 kJ/g, our measurement, unpublished).
[17_TD$IF]g
For nutrient sufficient conditions.
[18_TD$IF]h
For nutrient deplete conditions (low [19_TD$IF]nitrogen or phosphorous; silicon can also be depleted in diatoms (not listed here). Under these conditions,
suboptimal growth conditions (not reported in this table) are expected [4].

which can be compared with conventional biofuels: 120 for achieved because they are for multiple reasons
corn grain and 61255 for switchgrass or mixed prairie inherently more efficient solar energy converters.
grasses. This 612 fold increased energy yield for AMOPs Firstly, the intrinsic solar energy conversion efficiency
is one of several advantages they offer as sources of biomass (ECE)1 is greater for AMOPs at 39% (observed in the
and biofuel precursors relative to terrestrial plants. field) [12,13] versus a theoretical maximum of 2.4% for
C3 crops (switchgrass) and 3.7% for C4 crops (maize)
Examples: The hypercarbonate-requiring cyanobacterium across a full growing season based on calculated solar
Arthrospira (Spirulina) maxima thrives in volcanic soda lakes radiation intercepted by the leaf canopy [14]. The
at pH 9.511 and up to 1.2 M sodium carbonate, conditions disparity in favor of AMOPs is even larger with
that prevent most competing microbes. It grows to high cell increasing solar intensity, where multiple photopro-
densities in open pond reactors with low microbial con- tection mechanisms intervene that differentially
tamination [7,8]. Related strains are grown for the health protect terrestrial plants by diverting absorbed light
food industry, where productivity rates reach 27 dry metric to heat. Water and thermal stresses play major roles in
tons/ha  yr (annual in Mexico). Heated outdoor open exacerbating photoinhibition in terrestrial plants but
ponds have demonstrated annual yields of 6070 dry metric are much less important in AMOPs [15,16]. Secondly,
tons/ha  yr (in Israel) (Table 1). A salt-tolerant green algal AMOPs thrive across a greater range of light
strain of the genus Tetraselmis originating from the Great environments, with respect to mean photon flux, than
Salt Lake thrives at 6% salt, and a marine strain of this genus do higher plants. Thirdly, AMOPs are full canopy
has been cultivated in open ponds in New Mexico with absorbers, having superior light capture efficiencies
productivity rates of 38 (annual) and 69 (summer) dry when integrated over their growth cycle (13 dou-
metric tons/ha  yr (Table 1). This value increases to blings day 1) compared with slower growing, sparsely
146 if grown in shallow outdoor flume bioreactors in sum- seeded plants that attain full canopy for only a fraction
mer months in Hawaii [9]. Several reports have corrobo- of the year. Depending on the cultivation strategy,
rated considerably higher yields in closed, thermostated strain selection and location, AMOPs can be used to
photobioreactors with optimal light conditions. harvest solar energy using a dedicated culturing
footprint year-round, in contrast to the majority of
Several aspects of AMOP physiology are relevant for current agricultural crops. Improvement in ECE is one
evaluating their possible integration into renewable bio- area of research needed for all photosynthetic crops.
fuel applications [10,11]: Preliminary research in this regard suggests that ECE

1. Superior solar energy yields. The 612 fold energy yield 1

Measured as energy released by complete combustion of biomass
advantage of AMOPs versus terrestrial plants can be divided by the solar energy absorbed.

Current Opinion in Biotechnology 2008, 19:235240 www.sciencedirect.com


can be enhanced by reducing chlorophyll antenna size
for more efficient light utilization [17,18].
2. Lack of recalcitrant biopolymers. AMOPS are buoyant
aquatic microcells that do not require structural
biopolymers essential for higher plant growth in
terrestrial environments. The resulting absence of
cellulose, hemi-cellulose, and lignin eliminates the
need for pretreatments to breakdown cellulytic
products (hydrolysis by acid and enzymatic cleavage
by cellulase) and lowers the reactor temperature of
their subsequent fermentation. This simplification of
process and improved conversion efficiency bypass the
most costly and inefficient steps for conversion of
cellulose to ethanol [19]. Lignin cannot now be
readily transformed to transportation fuels. This
advantage of simpler biomass composition needs to
be weighed against the added cost for removal of
excess water in cases where, for example, pre-
processing for lipid extraction requires dry cell mass.
3. Metabolic and ecological diversity. AMOPs exhibit
enormous ecological, genotypic, and metabolic diver-
sities with over 4000 distinct species of cyanobacteria
(prokaryotes) and a comparable number of unicellular
algae (eukaryotes) classified thus far. This diversity
allows selection of genera/species that are adapted for
growth in locally available aquifers (marine, hypersa-
line, thermophilic and freshwater), or have morpho-
logical features that allow cost-effective harvesting
(filamentous, buoyant, or aggregate), or possess
anaerobic metabolisms that enable production of
hydrogen, ethanol, and/or organic acids by auto-
fermentation before final biomass conversion/utiliz-
ation. Moreover, several AMOPs are well characterized
and advanced genetic techniques exist for engineering
of strains with already demonstrated progress toward
optimizing bioenergy production and identifying
promising targets by genomic techniques [20].
4. Biosynthetic control of chemical composition by nutrient and
environmental stresses. The ability of AMOPs to direct
the majority of photosynthetic reductant into meta-
bolic pathways that synthesize the most amenable
bioenergy precursors starch, glycogen, and lipids rather
than cellulose and lignin offers a potential processing
simplification and gain in the overall energy yield.
Most AMOPs are rich sources of proteins when grown
under nutrient replete conditions (Table 1). Unlike
higher plants, they can be easily induced to alter their
composition using nutritional or environmental stres-
ses. Using these techniques, protein content can be
converted to energy storage compounds such as
carbohydrates (starch, glycogen, polysaccharides) or
higher energy lipids (fatty acids, monoacylglycerols,
diacylglycerols and triacylglycerols) in a species-
dependent manner. Many microalgae have the ability
to produce substantial amounts (e.g. 2050% dry cell
weight) of triacylglycerols (TAG) as a storage lipid
under photo-oxidative stress or other adverse environ-
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Aquatic phototrophs Dismukes et al. 237
mental conditions [10]. Lipids contain twice the
energy stored per C atom than do carbohydrates, which
translates directly into a twofold increase in fuel
energy content. Depletion of nitrate (N), silicate (Si)
or phosphate (P) from the growth medium has been
shown to produce twofold to threefold increase in the
relative amounts of starch (glycogen in cyanobacteria)
or lipids in many AMOPs [4]. This shift in
composition enables a growth strategy for enriching
cells in specific energy precursors before cell harvest-
ing. By judicious selection of the initial nutrient
concentrations for growth, batch culturing can take
advantage of this transition from rapidly-growing,
protein-rich young cells to mature cells that are richer
in biofuel precursors.
5. Contamination. The NRELASP project found that
contamination of non-native AMOPs by native AMOP
strains is a serious problem facing large-scale cultiva-
tion in open reactors [4]. Their high nutritional value
also makes them easy targets for predation. Suggested
improvements require the selection of robust strains of
AMOPs that thrive in high salt concentrations or high
pH where microbial contamination is low. These
conditions are unsuitable for many bacterial and fungal
contaminants [4,5]. Common natural aquifers that
harbor robust AMOP communities include the Great
Salt Lake and volcanic soda lakes. One such example,
A. maxima, possesses the highest solar energy conver-
sion of any measured oxygenic phototroph (AMOPs
and terrestrial plants) [21]. Much of this energy is not
used for biomass accumulation, but rather for excess
ATP production essential for maintaining osmotic
balance at high ionic strength. Consequently, such
environments represent a trade-off between pro-
ductivity and species competition.
6. Water usage and harvesting. Table 1 summarizes the
water usage per unit weight biomass and per energy
unit produced after annual recycling for AMOP
cultivation based upon commercial scale open pond
reactors. The volume is comparable or lower than that
required to grow maize at an equivalent weight of corn
grain, eightfold less than rape cultivation on fertilized
land, but 610 fold more than needed by unfertilized
switchgrass on non-arable land. This highlights a main
limitation of AMOPS in comparison with switchgrass.
The natural water source for many AMOPs is seawater,
which is plentiful and can be substituted by deep
saline aquifers in the interior [22] or nutrient-laden
agricultural wastewater [23], while terrestrial biofuel
crops require freshwater. Mass cultivation of AMOPs is
currently done at coastal locations in open ponds and
interior locations in desert climate using nutrient-
supplemented seawater where evaporative losses
dominate. Scale-up is under consideration using off-
shore sites bounded by wind-farm pylons. AMOPs
grown in hypersaline aquifers impose additional
constraints on water. However, the additional high
Current Opinion in Biotechnology 2008, 19:235240
238 Energy Biotechnology
Figure 1
Areas needed for cultivation of three biomass sources. Each box represents the area needed to produce a sufficient amount of biomass to convert to
liquid fuel to displace all gasoline used in the USA (2006 figures) on an energy basis. Data taken from ref 24.
salt and bicarbonate concentrations needed for their
culturing could be obtained from the concentrated
brine discarded during desalination of seawater. Water
desalination facilities are in high demand globally and
projections forecast escalating expansion [22]. The
synergism of these resources should be an important
consideration for future co-development of water
desalination and biofuels from AMOPs.
Harvesting of mass-cultured AMOPs constitutes a
major technical challenge for unicellular types that do
not aggregate. These cell morphologies require some
form of chemically induced coagulation or forced
flotation using compressed air bubbles that are costly
or have environmental trade-offs [23]. However, many
strains of AMOPs are filamentous that enable
separation by simple straining, or contain gas vacuoles
to aid in buoyancy, which allows isolation by
skimming. For this reason this class of AMOPs is
often chosen for commercial farming. Genetic tools are
limited for filamentous AMOPs and need to be
developed to allow this class of easy-to-harvest cell
types to be more fully exploited for bioenergy farming.
7. Valuable by-products. A significant fraction of the
residual biomass following lipid and carbohydrate
extraction is protein. This is expected to pass through
largely unaltered by the mild conditions used for
fermentation to ethanol and possibly those for lipid
Current Opinion in Biotechnology 2008, 19:235240
extraction for biodiesel conversion. Taking a holistic
view of AMOP energy processing, these by-products
can be directed toward secondary markets, such as use
of residual protein as animal feed (and mineral ashes in
the case of gasification).
Land versus aquatic biofuels
Figure 1 compares the areas needed for three biomass
sources. Data for corn grain and switchgrass/mixed prairie
grasses [24] are compared with AMOPs. Each box
represents the area needed to produce a sufficient amount
of biomass to produce liquid fuel to displace all gasoline
used in the USA (2006 figures) on an energy basis. For corn
and switchgrass, carbohydrate/cellulose content would be
fermented to ethanol. This estimate does not include the
requisite energy input for growth or ethanol production
(about 80% of the energy in corn ethanol, assuming an
optimistic figure). The switchgrass area assumes yet unspe-
cified optimal technology for both the production of cellu-
lytic feedstock and its subsequent fermentation to ethanol.
For AMOPs, either fermentation to ethanol or conversion
to biodiesel is an option depending upon species. We show
two boxes at 30 and 70% to illustrate the upper and lower
ranges expected for the conversion efficiency and relative
energy content of diesel versus ethanol. The overall solar
energy conversion to biofuel works out to about 0.05% for
www.sciencedirect.com

solar to ethanol from corn grain and roughly 0.5% for
switchgrass to ethanol [24]. Comparatively, this value is
about 0.51% for AMOPs to ethanol or biodiesel (ignoring
the fossil fuel input in all cases).
Conclusions
To realize the potential of AMOPs, significant research
efforts are underway to overcome several bottlenecks
hindering their widespread use, which include (a) cell
harvesting, (b) culturing strategies that maintain relative
monocultures and promote high photosynthetic conver-
sion efficiencies, (c) metabolic control (either physiologi-
cally or genetically) of the accumulated biopolymers, (d)
access to suitable aquifers, and (e) advanced biorefining
techniques to isolate biofuel precursors in a cost-effective
manner. Although obstacles to deployment exist, AMOPs
offer a wide range of alternatives for designer biomass
and biofuel precursors, and research into the use of
AMOPs as viable bioenergy feedstocks is in its infancy
relative to more thoroughly investigated terrestrial feed-
stocks. They are inherently more efficient solar collectors,
use less or no land, can be converted to liquid fuels using
simpler technologies than cellulose, and have secondary
uses that fossil fuels do not provide. Current research
efforts into the use of AMOPs as bioenergy feedstocks
will continue to leverage several of the assets inherent in
these organisms. The coming years will probably provide
new insights into novel ways to use AMOPs in economi-
cally viable biofuel processes.
Acknowledgements
The authors thank Mike Seibert for help with a figure, Bob Williams and
Lee Lynd for stimulating discussions, and Patricia Brletic and Lindsay
Leone for collaboration with combustion calorimetry experiments. GCD
and MCP research is supported by an AFOSR-MURI grant.
Appendix A. Supplementary data
Supplementary data associated with this article can be
found, in the online version, at doi:10.1016/j.cop-
bio.2008.05.007.
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