Wiratan 1

Karen Wiratan
16 January 2017
Period 3
Synthesis Paper

No matter how green we claim to be, the average American alone releases about eighteen

thousand kilograms of carbon dioxide every year. The percentage of carbon dioxide in our

atmosphere has shown a dramatic increase from around 367.5 ppm in April 2002 to 405.6 ppm in

November 2016. Forty seven percent of all carbon dioxide emissions come from both the

industrial and transportation industries in 2014 ("Total U.S.," n.d.). This neither bodes well for

our atmosphere nor our fast-speed society. In light of these problems, a new urgency arises for

the creation of alternative fuels in order to combat the continuous, rapid depletion of fossil fuel

reserves. Many energy sources, such as hydroelectricity and green fuels have been created and

scientific research has led up to the creation of fifth generation biofuels. Even with these

advancements, however, no alternative fuel source has become efficient and reasonable enough

for widespread commercialization. Microalgae has been recently recognized as holding great

potential for mass production, but many problems, such as the high costs associated with the

process of fuel conversion and the inefficiency of algae grown in crowded conditions currently

does not allow microalgal biofuels to reach their full capacity in production. In order to combat

the continuous, rapid depletion of fossil fuel reserves and inefficiency of microalgal biofuels,

metabolic engineering, a technique rising in potential to increase production, should be utilized

in the production process of these fuels in order to better the rate of lipid accumulation in

microalgae.

Microalgae have a unique metabolic structure that sets it aside from conventional

biocrops and other forms of biofuels. In addition to being a third generation biofuel, in its native

environment, microalgae are already highly productive (Li et al., 2008). The total amount of

solar energy captured by microalgae is approximately ten to twenty percent higher than that of
 Wiratan 2
Karen Wiratan
16 January 2017
Period 3
other terrestrial plants (2008). The simple structure of microalgae has led to a faster growth rate,

even allowing it to have the potential to be fifty times more productive than the biofuel crop

switchgrass, which is the fastest growing terrestrial plant (2008). Furthermore, triacylglycerides,

TAGs, are among the most abundant non-polar lipids in microalgae. Even when production

becomes slow, microalgae may potentially enable rapid adaptive membrane reorganization

(Sharma et al., 2012) to remedy the decrease in production. The production of microalgal

biofuels will not require much space or materials while also helping to improve the environment.

Unlike conventional biocrops, microalgae are able to endure unfavorable environmental

conditions due to TAGs, which acts as storage function (2012). Microalgae also do not require

nearly as much water as do terrestrial crops (Li et al., 2008). These factors allow the agriculture

industry face a decreased amount of competition for the same materials needed to cultivate food

and fuel crops. As microalgae can survive in more adverse environments, it also has a high

tolerance of carbon dioxide, thus allowing it to mitigate carbon dioxide flue gas. Along with this,

the metabolic pathways of microalgae may be manipulated to further increase the yield of useful

products due to the flexibility of the organism. For example, electromagnetic fields can stimulate

both growth and metabolic cascades, also being able to control biochemical pathways. The

conditions in which the microalgae grow in may help change the structure for the betterment of

production. With the plasticity of metabolic structure, scientists are working to find more

effective ways in order to increase yield.

Nutrient deprivation is a prominent technique in the metabolic engineering of microalgae

to increase the yield of products consumed for the production of microalgal biofuels. While this

technique places stress on the cell from the lack of necessary nutrients, it has been proven that

this does not hinder growth rate or biomass production. In a study of antisense-containing
 Wiratan 3
Karen Wiratan
16 January 2017
Period 3
knockdown strains, breeds of bacteria with certain inoperative genes, the growth rate is shown to

have decreased in comparison to natural microalgal strains (Trentacoste et al., 2013), but an

increase in lipid production. In another study involving microalgal species T. weissflogii and C.

cryptica, the amount of TAGs increased under nutrient deprivation (dIppolito et al., 2015).

Nutrient deprivation causes the up-expression of genes. Genes responsible for catalyzing carbon

dioxide and ammonia were up-expressed for a while under nutrient deprivation. Genes relating to

the production of fuels were also shown to have been up-expressed. In another microalgal

species, P. tricornutum, genes that participate in the utilization of nitrate and phosphate had been

up-expressed.

This technique allows for not only the up-expression of genes, but the utilization of less

commonly used metabolic pathways. Under unfavorable conditions, pathways can be

reprogrammed under stress to produce more lipids, which consist mainly of neutral lipids that

have a great majority of TAGs. Biomass and TAGs compete for resources and a greater

percentage of production (Sharma et al., 2012). Pathways may be engineered to make the process

even more efficient by decreasing the number of steps needed. For example, nutrient deprivation

can facilitate the production of hydrogen (Wilhelm & Jakob, 2011). This can then make the

process more cost-effective, which is currently a main obstacle separating microalgal biofuels

from commercial production. As nutrient deprivation has shown great potential for allowing the

commercialization of microalgal biofuels, it is also important to find the most effective technique

within nutrient deprivation in order to optimize the productivity.

Nitrogen deprivation is one of the most widely used of the stressors in nutrient

deprivation. This technique allows for a more efficient process of nutrient deprivation in

comparison to other widely used methods. Out of other methods, it is the most effective
 Wiratan 4
Karen Wiratan
16 January 2017
Period 3
induction technique. The low amount of lipids triggered an increase in the production of TAGs, a

type of nonpolar lipid, while the production of polar lipids decreased (dIppolito et al., 2015). An

increased amount of light is absorbed by microalgae, thus further increasing productivity. In a

study done in 2008, it was stated that[P]arietochloris incise grown in nitrogen-replete medium a

considerable increase in the ratio of carotenoid and chlorophyll contents was recorded (Li, et

al., 2008). That is, Parietochloris, a genus of microalgae was recorded to have a significant

growth in the amount of carotenoids and chlorophyll when nitrogen was depleted. This allows

for an increase in the absorbance of solar energy, as carotenoids and chlorophyll are both

pigments whose function is to effectively capture light, which would lead to a higher production

of lipids.

Nitrogen is the nutrient with the greatest impact on lipid accumulation in microalgae. The

effect nitrogen has on metabolic structure includes leading to a different process to synthesize

TAGs. In P. Tricornutum, the production of TAGs was increased under the stress of nitrogen

deprivation (dIppolito et al., 2015). Nitrogen deprivation also causes a remodeling of the lipids

produced. Above 80% of glycerolipid production consisted of TAGs in T. weissflogii and C.

cryptica (dIppolito et al., 2015, para. 11). When used in conjunction with other techniques,

especially, the efficiency of nitrogen limitation may be increased. An important technique to be

noted when used with nitrogen deprivation is the use of hydrogen peroxide treatment. The ratio

of neutral lipids to biomass was measured to have increased by greater than fifty percent (Burch

& Franz, 2015). Along with lipid accumulation, waste conversion may be optimized to not only

help in reducing carbon dioxide emissions along with other harmful substances, but furthermore

the efficiency of biofuels (Hnain et al., 2011). Along with hydrogen peroxide, interactions
 Wiratan 5
Karen Wiratan
16 January 2017
Period 3
between microalgae and biofilms have shown a higher tolerance to ethanol, higher yields, and a

more efficient continuous system that permits separation and extraction (2011).

As the quality of our air, resources, and environment continues to diminish, the need for

green technologies quickly becomes greater. Microalgae have been recently recognized in the

scientific community as a good potential candidate for an alternative to fossil fuels due to its high

efficiency rate in nature and in laboratories. In order to fully take advantage of this discovery,

several techniques have been explored in order to find the most efficient in increasing yield. Due

to the unique traits and flexibility of microalgaes metabolic structure, metabolic engineering has

become one of the more popular choices in laboratories dealing with the algae. Unlike

conventional biofuels, microalgae are able to thrive in more adverse environments, thus reducing

interference with other industries for resources and the cost of supplies. In addition, it has been

observed by many experiments that the nutrient deprivation of microalgae triggers the increased

production of TAGs, a fatty acid necessary for the production of microalgal biofuels. More

specifically, nitrogen deprivation has been more commonly used and seems to be a very effective

stressor of nutrient deprivation. The commercial production of microalgae relies heavily on the

optimization of yield in order to obtain a higher efficiency; otherwise, it will not be able to

compete with other fuels on the market. Many hurdles still lie in the path to the widespread

commercialization of alternative fuels, but, with metabolic engineering and the potential of

microalgae, microalgal biofuels may come to be the first biofuel to become a main source of

energy.
 Wiratan 6
Karen Wiratan
16 January 2017
Period 3

References
Burch, A. R., & Franz, A. K. (2915, August). Combined nitrogen limitation and hydrogen
peroxide treatment. Davis, CA: Biosource Technology.
Carbon Dioxide. (2016, November). Retrieved January 4, 2017, from NASA Global Climate
Change website: http://climate.nasa.gov/vital-signs/carbon-dioxide/
dIppolito, G., Gallo, C., Botte, P., Fontana, A., Sardo, A., Vella, F. M., . . . Paris, D. (2015,
February). Potential of lipid metabolism in marine diatoms for biofuel production.
Retrieved from https://biotechnologyforbiofuels.biomedcentral.com/articles/10.1186/
s13068-015-0212-4
Hnain, A. K., Cockburn, L. M., & Lefebvre, D. D. (2011, March). Microbiological processes for
waste conversion to bioenergy products: Approaches and directions. Kingston, Canada:
NRC Research Press.
Li, Y., Horsman, M., Wu, N., Lan, C. Q., & Calero, N. D. (2008, July). Biofuels from
Microalgae. Retrieved from Wiley Online Library website:
http://onlinelibrary.wiley.com/doi/10.1021/bp070371k/full
Peacock, S. (2014, October 2). How much CO2 are you emitting? [Blog post]. Retrieved from
Earth on the Edge website: http://www.earthontheedge.com/
how-much-co2-are-you-emitting/
Sharma, K. K., Schuhmann, H., & Schenk, P. M. (2012, May). High Lipid Induction in
Microalgae for Biodiesel Production. Retrieved from
http://www.mdpi.com/1996-1073/5/5/1532/htm
Trentacoste, E. M., Shreshta, R. P., Smith, S. R., Gl, C., Hartmann, A. C., Hildebrand, M., &
Gerwick, W. H. (2013, December). Metabolic engineering of lipid catabolism increases
microalgal lipid accumulation without compromising growth. San Diego, CA:
Proceedings of the National Academy of Sciences of the United States of America
PNAS, Proceedings of the National Academy of Sciences.
Total U.S. Greenhouse Gas Emissions by Economic Sector in 2014 [Chart]. (n.d.). Retrieved
from https://www.epa.gov/ghgemissions/ sources-greenhouse-gas-emissions