INTRODUCTION

Petroleum, coal, and natural gas, these are three primary sources of global energy that are currently available. These non-renewable sources provide 90% of the total worlds energy needs (Dale 2008). Upto 40% of the energy needs are provided by petroleum oil (Sivakumar, Vail et al. 2010). Fossil fuels are formed from buried organic materials, include dead plants and animals, which have the geologic transformation over millions of years and these are non-renewable. The energy demand over the world has increased due to the rapid increase in the human population, rapid growth of the industry and the increase in the use of transportation. According to a recent report the consumption of fossil fuel is 105 times faster than nature can generate it

(Satyanarayana, Mariano et al. 2011). Fossil fuels are about to reach their peak production and can be predicted to be exhausted in the future due to their limited and non-renewable nature. Therefore, the current use of fossil fuels is widely recognized to be unsustainable. Another major concern with using fossil fuels is carbon dioxide (CO2) emission. Since the beginning of the Industrial Revolution, the burning of fossil fuels has been contributing to the increasing atmospheric CO2 concentration. Many scientists believe that fossil fuel derived CO2 is one of the major factors for greenhouse effect, which increases temperatures of global temperature. According to the data from U.S. Energy Information Administration, over 80% of the greenhouse gas emissions come from energy-related CO2 emissions in which petroleum is the largest fossil fuel source for 2

energy-related CO2 emissions, contributing 42% of the total (U.S. Energy Information Administration. 2011). Additionally, the high petroleum oil price is a big challenge for the 21st century, especially after the oil crisis in 1973 and then the Gulf War in 1991. Even though the price has fallen recently, petroleum fuel price will undoubtedly rise again because of the increasing energy consumption and limited availability. Worldwide concerns resulted in the search for new energy resources which would be renewable and sustainable. Bio-mass source fuel, or bio-fuels, may offer a promising alternative which have attracted increasing attention as evidenced by the growing

research and development efforts found in the literature (Solomon 2010; Durrett, Benning et al. 2008). Fuels are the basic need of current era to make everything working. A worldwide dependence on the products derived from fossil fuels has significantly increased the usage of these resources. Fossil fuels are considered to be non-renewable resources, requiring millions of years to form. This long period required to replenish these important natural resources is just one of the drivers in the hunt for alternative sources of energy. It is a U.S. national goal to develop sustainable practices for fuel production from renewable resources to reduce dependence on imported oil [1]. The United States is the top oil consumer in the world [2]. In 2009, the amount of oil produced in the U.S. was 9.14 millions of barrels per day and the amount consumed was 18.81 millions of barrels per day [2]. With the continual depletion of fossil fuels, the U.S.s consumption rate will no longer be possible unless a viable alternative energy source is found. The increasing cost is causing difficulty for not only consumers but also businesses. Services which require the usage of fuel become more expensive to operate resulting in an increased cost to the consumer. Furthermore, fossil fuels have a high carbon footprint, which determines total greenhouse gas (GHG) emissions. Therefore, it is important that alternative energy resources have a small carbon footprint and a beneficial cost impact to the end consumer. 2 Alternative energy sources include wind, solar, geothermal, hydroelectric, nuclear, and biofuels. Of these, biofuels alone can provide an alternative source of liquid transportation fuels. Biofuel is obtained from carbon-rich plant and animal biomass. Plants are renewable and utilize natural sources for growth; the sun, water and carbon dioxide (a GHG). The short growth period of plants relative to fossil fuels, make them a renewable energy source. Biomass can be used as feedstock to produce various types of biofuels depending on the carbon source used such as bioalcohols, biodiesel, green diesel, bio-oil and syngas. Biofuels are often referred to as first, second or third generation fuels. The first generation uses sugars or lipids derived from grain food crops: corn and soy being two large sources. Using these crops to produce biofuel can conflict with the food supply and pricing. The second generation of biofuels is derived from lignocellulosic biomass. These plants are non-food crops or inedible plant waste composed of the organic polymers; lignin, cellulose, and hemicelluloses. A large obstacle to overcome with woody biomass is the difficulty in accessing fermentable sugars.

Lignin can impede this access. Sugars are fermented to produce ethanol using technologies similar to those in corn to ethanol production. The third generation of biofuels is produced from algae. Algae are aquatic, autotrophic organisms that chiefly contain three important biopolymers; protein, carbohydrates and lipids. Almost all algae are free of lignin. Algal species do not compete with food sources and can use waste water and carbon dioxide for growth which could be recycled from algal processing waste. In a study from 1978 to 1996, the Aquatic Species Program, investigated the use of high lipid algae for biodiesel 3 that could utilize waste carbon dioxide from coal fired power plants [3]. In recent years, this research and development has been revived. The growth of algae can be more rapid than other biomass, allowing a fast turnaround for biofuel production. For most crops, the low number of harvests per year limits the yield of biofuel that can be produced. Mono-culture plots can also threaten biodiversity and can have a negative impact on greenhouse gas emissions when extensive soil fertilization is required [4]. Greenwell et al. also found that the yield of fuel per area for algae is theoretically greater than that of corn and other feedstocks, although a detailed life cycle assessment (LCA) of sustainability must be developed. A 2011 review on LCA methods concludes that there are many variables involved in the conversion of biomass to biofuels, making it difficult to make complete assessment of the viability and advantages of biofuel over fossil fuel [5]. This leads to difficulty in determining the most promising type of biomass for the replacement of transportation fuel and other energy sources. It is known that to be a practical alternative, a biofuel should provide a net energy gain, have environmental benefits, be economically competitive, and be producible in large quantities without reducing food supplies [6]. These factors are of utmost importance when evaluating any biomass as a potential feedstock for biofuel production. Algae are thought to be potentially viable feedstock for fuels, motivating research and development efforts for its use in alternative energy. Biodiesel and jet fuel can be produced from lipid-rich algae. Lipid processing techniques include transesterification to convert lipids to alkyl esters (biodiesel), refining free fatty acids (FAs) or esters by catalytic cracking and hydrotreating for jet fuel production, or the UOP/Eni Ecofining 4

process to obtain clean green diesel [7]. As with corn and other cellulosic biomass, carbohydrates from algae can also be used to ferment sugars for bioethanol, a bioalcohol, production. Algal proteins can be useful as livestock feed or a food supplement. Processing these important constituents utilizes the entire algal product. Green processing of algal lipids by hydrolyzing triglycerides to FAs using high temperature (subcritical) water is the motivating goal for this study. The oil phase consisting of FAs, free of glycerol and water, can be directly converted to high quality biofuels. The use of water as a solvent eliminates the need for organic solvents and an external catalyst. The aqueous hydrolysis product containing glycerol could be a useful growth media for the algal culture, adding sustainability to processing. Protein extraction prior to lipid hydrolysis avoids charring and thermal degradation of useful proteins. Consequently, increased efficiency of hydrolysis and the ability to produce protein bioproducts can be achieved. Biodiesel is a fast expanding industry within the United States that is facing a growing dilemma: a feedstock source capable of keeping up with demand (Eriksen 2008). As energy demands skyrocket and oil reserves begin to falter in their stability, the need for a reliable renewable fuel source grows. There are many options in this area, but unlike solar, nuclear, and fossil fuels, biofuels such as bioethanol, biodiesel, and green diesel have the capability of providing a fuel source ideally suited to existing infrastructure within the transportation industry. Biodiesel, when blended with petroleum diesel, can be used in unmodified diesel engines. It has the added benefit of higher lubricity than petroleum diesel, so it helps provide for greater longevity within diesel engines. To use B100, or pure biodiesel, an engine usually needs a slightly modified fuel rail so prevent fouling, but such modifications are inexpensive when compared to other changes necessary to adopt competing forms of transportation fuel or energy (Chisti 2007; Chisti 2008). Fertilizer run-off on farms across the United States is causing large-scale problems among local streams and tributaries, spreading to river networks, and also affecting the coastal areas and bays. Algae blooms caused by nutrient leakage can often be traced to large scale

operations that produce crop or animal products (Craggs, Adey et al. 1995; Kebede-Westhead, Pizarro et al. 2006). Excess fertilizer provides food for algae in these situations and the algae take over a local part of the environment, causing declines in the population of native species. Algae blooms are simply one result of nutrient runoff. Often, the high nitrogen and phosphorous contents within the runoff is enough to negatively affect local fauna populations within streams, and it can impact drinking water as well. Macroalgae has been used in the past as a way to recycle some of the nutrients within these wastewater sources, and also as a step in industrial wastewater treatment (Pizarro, Kebede-Westhead et al. 2002; El Sikaily, Khaled et al. 2006). Utilizing this otherwise harmful product as a food source for microalgae capable of producing biodiesel is a potential solution. Added benefits include nutrient recycling, and the reduced cost of providing a growth medium for the algae. 2 Currently, algal biofuel production has not been commercialized due to high costs associated with production, extractiontransesterification harvesting, and preparation for traditional

into fatty acid methyl esters (FAME). The reaction to make biodiesel, once the lipids from the algae are extracted, is the same as when using virgin vegetable oils, which has been in use for a number of years (Apt and Behrens 1999). This reaction is known as transesterification. However, most methods used to extract the oil from algal biomass rely on a dry biomass product. Previous research has usually used lypholization to ensure low homogenous moisture content in the cells (Li, Horsman et al. 2007; Li, Du et al. 2008; Refaat, Attia et al. 2008). This is not very practical in industry due to its high cost and slow dehydration times. Oven drying in industry is also not ideal, as it can lead to damage of the lipid feedstock. Using temperatures low enough to protect the lipids causes an increase in drying time, and other methods, such as forced air drying, cause an increase in the price. These excess costs associated

with biomass pretreatment are what cause the feedstock costs to be the highest expense associated with biodiesel production, and microalgae is no exception. Indeed, microalgal oil costs considerably more than soybean oil currently, due mainly to the existing infrastructure for terrestrial crops and the lack of investment in the culture of large scale microalgal aquatic organisms (Eriksen 2008).