Energy Convers. Mgmt Vol. 36, No. 6-9, pp. 707-712. 199.

5
Elsevier Science Ltd. Printed in Great Britain
0196-8904(95)00103-4

THE USE OF MICROALGAE FOR ASSIMILATION AND

UTILIZATION OF CARBON DIOXIDE FROM FOSSIL FUEL-FIRED

POWER PLANT FLUE GAS

KATHRYN G. ZEILER, DANA A. I-IEACOX, SUSAN T. TOON

KIRAN L. KADAM AND LEWIS M. BROWN

National Renewable Energy Laboratory

1617 Cole Blvd., Golden, Colorado, USA 80401

Absract - The accumulation of carbon dioxide in the atmosphere, primarily as a result of the
combustion of fossil fuels, has been linked to potential global climate change. Capture and
utilization of the carbon dioxide by microalgae has emerged as a promising technology to
help reduce emissions from fossil fuel-fired power plants. Microalgae are of particular
interest because of their rapid growth rates and tolerance to varying environmental conditions.
We are currently conducting experiments on the growth of microalgae exposed to simulated
flue gas, This technology is envisioned for open raceway cultivation ponds as a low cost
implementation strategy. Coupling the production of fuel or commodity chemicals with the
use of flue gas carbon dioxide as a microalgal nutrient is envisioned to be a cost-effective
method of reducing the amount of carbon dioxide contributed to the atmosphere by fossil
fuel-tired power plants.

Kev Words: carbon dioxide, microalgae, fossil fuel combustion, SO,, NO,

1. INTRODUCTION

Carbon dioxide is viewed as a potential agent of global climate change. Anthropogenic emissions of
carbon dioxide are estimated to be 2 x 10” tons/y, primarily from combustion of fossil fuels. In addition, the
increasing global population pushes the demand for economic energy sources higher every year. Contrasted
with its potential for harm is the value of carbon dioxide - it is the source of carbon for photosynthesis, without
which there would not be life on earth as we know it today. Carbon dioxide is a valuable resource for many
of man’s activities including enhanced oil and gas recovery, urea production, and food and beverage
carbonation.’ These activities and others of similar nature, however, can use only a very small percentage of
the total carbon dioxide emissions from fossil fuel combustion.

Using carbon dioxide from fossil fuel combustion as a feedstock for photosynthetic microorganisms
can provide a large sink for carbon assimilation. Microalgae are the most productive carbon dioxide users
with yields of biomass per acre threefold to fivefold greater than that from typical crop plant acreage’. Earlier
studies3-5 have discussed the potential applications of power plant flue gas to microalgal farming.
Experimental data are needed in order to determine the feasibility of this technology. The work presented here
discusses the current efforts at the National Renewable Energy Laboratory (NREL) toward deployment of this
technology and more specifically, laboratory bench-scale investigations into the ability of photosynthetic
microorganisms to assimilate carbon dioxide from flue gas.

707
708 ZEILER et al.: ASSIMILATION AND UTILIZATION OF CO? FROM FLUE GAS

2. CULTURE STUDIES

Initial studies were carried out using a green alga, Monoruphidium minutum (NREL Strain Monor02).
Particular emphasis was placed on the gas delivery apparatus, as efficient dosing of the flue gas is crucial to
successful growth Mass culture facilities typically provide carbon dioxide “on demand” as determined by
pH of the culture, i.e., they are operated as pH stats. This allows the acidic gas to neutralize the alkalinizing
effect of cell growth. Appropriate dosing allows efficient utilization of the introduced carbon; which is

107

106

4E 72 96 P0 144 48 72 96 120 144

timCh1 timeCh1

Fig. 1. Comparison of cultures exposed to either 1990 CAA simulated flue gas or control gas at 60 mL/min
(at SIP) three times per 24 h (left panel) or at 15 mL/min (at STP) 55 times per 24 h (right panel). These data
are representative of many experiments. The slight difference at 96 h is seen in some experiments and not in
others.

important since the efficiency of carbon dioxide utilization will have a significant impact on process
economics. Previous studies showed that exposing cultures to continuous dosing with simulated flue gas
resulted in no evidence of growth or viability of tbe culture6.

The experiments described here examine the effect of simulated flue gas containing 1990 US Clean Air Act
(CAA) levels of sulfur and nitrogen oxides on culture growth and biomass accumulation as a measure of
carbon assimilation. Cultures were inoculated at 2 105cells/ml into a fmal culture volume of 300 ml (10%
artificial sea water’). Light was provided continuously by Cool White@fluorescent bulbs at an intensity of
-200 uE m%‘. The cultures were continuously stirred with a magnetic stirrer at 150 rpm and the temperature
 ZEILER et al.: ASSIMILATION AND UTILIZATION OF CO? FROM FLUE GAS 709

was maintained at 25’C. The simulated flue gas contained 0.015% NO, 0.02% SO,, 5% 0,. 13.6% CO,, with
the balance N,. The control gas contained 13.6% CO,, 5% 0,. with the balance N,. The sparging regime was
varied depending upon the experiment.

In the initial experiments, the cultures were exposed to either simulated flue gas or control gas
containing an amount of carbon sufficient to acheive an estimated yield of -1 g dry wt biomass L‘l de’and
delivered in 3 equivalent doses during an 8 h time period each day. The flow rate in these studies was 60
mLJmin at STF’. Rate of growth, biomass accumulation, chlorophyll concentration, and pH were among the
parameters measured. In subsequent work the dosing rate was changed to determine if the carbon delivery
scheme was limiting culture growth. The same total amount of carbon was delivered; however, the flow rate
was decreased to 15 mL/min (at STP) in 2 min intervals 55 times during the 24 h period. Fig. 1 (above)
illustrates that there was no significant change in the rate of growth or total number of cells compared to the
initial delivery schedule, suggesting that the instantaneous carbon delivery rate is not limiting under these
conditions.

It is important to note that there is also no significant difference in the rate of growth between cultures
exposed to simulated flue gas and those exposed to control gas. These data indicate that 150 ppm NO and 200
ppm SO, are not inhibitory to culture growth under these conditions. Many of these experiments were carried
out for as long as 2 wks without evidence of inhibition of the culture. This is particularly encouraging because

8.0 ,

7.8 - 8 control gas

---e-- flwpaa

7.6 -

7.4 -

7.2 -
s
a 7.0 -

6.8 -

6.6 -

6.4 -

-I
48 72 96 120 144
time (h)

.x0, 7c.H

Fig. 2. The effect on culture medium pH upon the introduction of either simulated flue
gas or control gas. The pH was measured on samples taken during sparging.

an accumulation of acidic sulfur species in the media could cause inhibition of growth. Measurements of pH
during the course of the experiment indicate that pH rises as biomass increases, as shown in Fig. 2. Early in
the experiments, when biomass is low, the pH lowering effect of the exposure to the gases is more significant
even though the medium is buffered. The pH profile of media (that does not contain biomass) sparged with
simulated flue gas or control gas does not exhibit this increase in pH at later time points (data not shown).
710 ZEILER et al.: ASSIMILATION AND UTILIZATION OF CO, FROM FLUE GAS

Since the instantaneous carbon delivery rate did not appear to be limiting, we decided to determine
whether other key nutrients might be limiting culture growth. Nitrogen and phosphorus appeared to be likely
candidates as they are often limiting nutrients in natural populations.

In the next series of experiments, the gas delivery and all other experimental conditions were as
described above except that the nitrogen concentration was increased three-fold and the phosphorus
concentration six-fold (over the levels in
10% ASW). Fig. 3 shows the growth of
cultures exposed to high or low nutrient
levels. There was a dramatic increase in
cell number, biomass and chlorophyll
concentration observed when the level of
these nutrients was increased. The cell
number was nearly three-fold higher than
that observed in previous experiments
and the final biomass concentration was
significantly higher than that of cultures
given lower amounts of nitrogen and
phosphorus (see Table 1, below).
Chlorophyll levels in cultures exposed to
high nutrients were four-fold higher than
in cultures receiving the lower nutrient
levels. The impact of these findings is
that changes in the culture medium result
in significant increases in the amount of
carbon assimilated (as reflected by
fixation into biomass).
Studies in outdoor mass culture
ponds in New Mexico showed that
virtually 100% of the carbon dioxide fed
to the ponds was assimilated into
+ control gas. low N & P
-o- flue gas, low N & P biomas?. In these mass culture studies,
106
--D- control gas, high N &P carbon dioxide was delivered on demand
B --a-- flue gos, high N & P as determined by pH. Although urea and
5
phosphate were provided to these ponds,
3
the laboratory studies may identify
2 nutrient levels that will create an even
larger demand for feedstock carbon. In
105
the laboratory experiments at high
nutrient concentrations, all the nitrogen
0 24 48 72 96 120 144 168 192 21E was depleted by the end of the
time (h) experiment (data not shown). This
suggests that even greater carbon fixation
may be realized with further increases in
the levels of these nutrients.

Fig. 3. Comparison of the growth of cultures exposed to

either simulated flue gas or control gas and low or high
levels of nutrients (N and P).
 ZEILER et al.: ASSIMILATION AND UTILIZATION OF CO, FROM FLUE GAS 711

Table 1. Comparison of the effect of increased nitrogen and phosphorus on growth, biomass and chlorophyll
concentrations for cultures exposed to either simulated flue gas or control gas (sparging was at 15
mL/min(STP) for 2 min., 55 times per 24 h).

Low Nutrients High Nutrients

1.76 mM nitrate 5.28 mM nitrate
Table 1. 36 uM
. vhosvhate
_ 2 17 uM
. ohosohate
_ _

I
Measured Control Flue Gas Control Flue Gas
Parameter Gas Gas

Ash-free 1.32 1.25 1.88 1.70

Dry Weight +0.0006 kO.03 1 ‘kO.03 1 kO.0474
(g/L)

chlorophyll 7.40 9.08 39.17 41.39

(@ml) kO.087 fl.308 fl.662 kO.345

3. Outdoor Mass Culture Test Facility

NREL previously operated an Outdoor Test Facility (OTF) in Roswell, New Mexico to carry out an
engineering design assessment of microalgal mass culture in the US desert Southwest. Two 0.1 hectare ponds
were operated successfully, providing a great deal of information regarding microalgal growth rates in mass
culture, carbon dioxide utilization, and pond design parameters*.

As discussed above, during operation of the OTF, virtually 100% of the delivered carbon dioxide was
absorbed, demonstrating the practicality of this design. In addition, we have recently begun an in-depth
characterization of resource availability with respect to land, water, incident radiation, and similar factors to
assess implementation of this technology on-site at a coal powered electrical utility plant in Water-flow, New
Mexico. Preliminary consideration of suitable land and water resources in this area suggest they are available
at a reasonable cost.

4. Potential Products and Genetic Engineering

One use of the biomass produced by microalgal mass culture using the carbon dioxide from flue gas
is co-combustion for energy generation. Although CO, would still be released to the atmosphere, there would
be an overall reduction in the amount per unit of energy produced as the microalgae would essentially recycle
the carbon from the power plant stack. Alternatively, the biomass can be used to produce transportation fuels
(such as biodiesel) or commodity chemicals, again acting to decrease the carbon dioxide generated by the use
of petroleum products. All of the suggested scenarios would serve to lower the nation’s dependence on
imported oil.

Recent research efforts have concentrated on applying genetic methods to microalgae’*” in order to
develop organisms optimized for high productivity and energy value. DNA from microalgae has been
analyzed chemically”, and transient expression of a foreign gene (firefly luciferase) was demonstrated12.
Progress has also been made with respect to the development of new protocols for introducing foreign genes
into microalgae.’ In the area of microalgal lipid biochemistry and molecular biology (important for
production of biodiesel), an important enzyme in lipid metabolism, acetyl-CoA carboxylase was purified.‘4
This purified enzyme aided efforts to clone and sequence this key gene in lipid metabolism.‘5 Genetic
engineering studies should result in organisms with desired traits, thereby improving process feasibility.
 ZEILER et al.: ASSIMILATION AND UTILIZATION OF CO, FROM FLUE GAS
712

5. Conclusions

Combining microalgal farming with fossil fuel energy production has great potential to diminish
overloading of the atmosphere with carbon dioxide, as well as for production of useful products (i.e., energy
from combustion of biomass, transportation fuels such as biodiesel or commodity chemicals). The laboratory
studies described here demonstrate that microalgae can efficiently utilize simulated flue gas containing high
levels of carbon dioxide, as well as sulfur and nitrogen oxides, as a feedstock to produce substantial biomass.
Several parameters important for productivity have been identified including nutrient levels and sparging
regime. Operation of the mass culture facility in New Mexico demonstrated that there are no significant
engineering barriers to large scale pond culture. Other workers 16*17 have demonstrated direct utilization of
actual flue gas in demonstration-scale ponds containing sea water by a naturally occurring algal species.
These studies illustrate that implementation of this technology is eminently feasible and that while further
investigation into culture physiology and other areas will refine process design, no major technological
breakthroughs are needed.

6. References

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4. P. L. Chelf, L. M. Brown and C. E. Wyman, Biomass and Bioenergy Vol. 4, No. 3, 175-183 (1993).
5. E. A. Laws and J. L. Beming, Biotech. and Bioengineering, Vol. 37,936-947 (1991).
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and K. G. Zeiler, ACS Symposium Series 566:255-270 (1994).
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