Algae Technology Report

Index: The conversion of CO2 P. 2 Preferred Algae Strain P .2 Carbon fixation in Algae P. 2 Photosynthesis P. 4 Synthesis of carbohydrates P. 7 Photosynthetic yield and energy conversion efficiency P. 8 Algae as animal feed P. 9 Algae as a substitute feedstock for cattle P. 11 Bioreactors P. 12 Photo-bioreactors P. 13 Algaenius Photo-bioreactor design P. 13 High efficiency light source P. 14 Agitation and gas mixing P. 15 Control systems P. 16 Proposed PBR layout P. 17

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Conversion of CO2 to a useful carbohydrate While CO2 is a fairly stable molecule, it is the basis for the formation of complex sugars (food) by green plants through photosynthesis. The relatively high content of CO2 in flue gas (approximately 140,000 ppm for a coal-fired plant compared to the 350 ppm in ambient air) has been shown to significantly increase growth rates of certain species of microalgae by a factor of 300. Through natural photosynthesis, the algae will grow in volume while absorbing most of flue gas pollutants (i.e. carbon dioxide, nitrogen oxides, etc) and then converting them into biomass and thus essentially recycling pollution. The chemical reaction traps the carbon in the algae and gives off oxygen and nitrogen, which by itself is a normal component of air. Given that the average composition of Green Algae (normalized with respect to carbon) is CH1.8N0.17O0.56, then one mole of CO2 is required for the growth of one mole of 3 microalgae. Based on the relative molar weights, the carbon from 1 kg of CO2 could produce increased microalgal mass of 25/44 kg, with 32/44 kg of O2 released in the process, assuming O2 is released in a one-to-one molar ratio with CO2. Therefore, a photosynthetic system provides critical oxygen renewal along with the recycling of carbon into potentially beneficial biomass. The preferred algae strain While some algae strains are utilized in the bio-oil industry specifically due to their lipid production rate, others are chosen for their starch and / or biomass content. Algaenius has selected a strain that not only has all of these features but it also contains an enzyme that allows Carbonic Anhydrase (CA). The CA enzyme assists in the rapid inter-conversion of carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. Given that additional protons are liberated during this reaction, even more CO2 can be metabolized. This rate of metabolization normally increases the efficiency of a photosynthetic system by up to 300 times more efficient in terms of productivity: Reaction catalyzed by carbonic anhydrase: CO2 + H2O Carbonic Anhydrase HCO-3 + H+ The reaction rate of carbonic anhydrase is one of the fastest of all enzymes, and its rate is typically limited by the diffusion rate of its substrates. The important fact about the CA enzyme, is that while it can drastically increase the efficiency of the reaction, the CA assisted photosynthetic process requires a very specific light spectrum range and growth environment. Carbon fixation in Algae Carbon fixation is a process found in autotrophs (organisms that produce their own food) whereby carbon dioxide is changed into organic materials and are utilized in the metabolic pathway of photosynthesis. This process converts carbon dioxide and ribulose bisphosphate (RuBP, a 5-carbon sugar) into 3-phosphoglycerate. It is
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important to note that two 3-phosphoglycerate are produced for each molecule of CO2. Ribulose-1,5-bisphosphate carboxylase / oxygenase, (RuBisCO) is an enzyme that is used in the Calvin cycle to catalyze the first major step of carbon fixation, a process by which the atoms of carbon dioxide are made available to algae in the form of energy-rich molecules such as sucrose. RuBisCO catalyzes the carboxylation of the 5-carbon sugar ribulose-1,5-bisphosphate (RuBP) with carbon dioxide. RuBisCO is very important in terms of biological impact because it catalyzes the most commonly used chemical reaction by which inorganic carbon enters the biosphere. RuBisCO is apparently the most abundant protein on Earth and given its important role in the biosphere, there are currently efforts to genetically engineer crop plants so as to contain more efficient RuBisCO. Plants and algae that survive solely on carbon fixation tend to thrive in areas where sunlight intensity is low, temperature is moderate, water is plentiful and CO2 concentration is high. Plants and algae that utilize carbon fixation most likely originated during the Mesozoic and Paleozoic periods (100 to 600 millions of years ago) where the CO2 levels were extremely high (see below in parts per million) and allowed the plants and algae to flourish.

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Unicelluar algae are usually found in areas with high concentrations of CO2, otherwise the algaes RuBisCO will incorporate an oxygen molecule into the RuBP, instead of a carbon dioxide molecule. This breaks the RuBP into a three-carbon sugar that can remain in the Calvin cycle, wasting the cell's energy. High concentrations of carbon dioxide lower the chance that RuBisCO incorporates an oxygen molecule. Most other plants have adaptations that allow them to survive in areas where the plant cannot take in a lot of carbon dioxide. RuBP is an important substrate involved in carbon fixation. The enzyme RuBisCO catalyzes RuBP with CO2 in order to synthesize a highly unstable 6- carbon intermediate known as 3-keto-2-carboxyarabinitol 1,5-bisphosphate, which decays virtually instantaneously into two molecules of glycerate 3- phosphate (3PG) (see equations below). RuBP is both used and created during the Calvin Cycle. It is a product of the phosphorylation of ribulose-5-phosphate by ATP. This extremely unstable molecule created by the initial carboxylation was unknown until 1988 when it was isolated. The 3-phosphoglycerate can be used to produce larger molecules such as glucose. Carbon fixation equation: a) 5-carbon RuBP + 1-carbon CO2 b) Which forms 6-carbon intermediate c) Then forms two 3PG d) Each 3PG (3-carbon) is then reduced to G3P in carbon reduction e) Five G3P = 3 RuBP f) Two G3P = 1 C6H12O6 (glucose) Some enzymes typically can carry out thousands of chemical reactions each second. However, RuBisCO is slow, being able to "fix" only 3 carbon dioxide molecules each second. Nevertheless, because of its extremely large concentration and even when light is otherwise limiting photosynthesis (RuBisCO is usually only active during the day because RuBisCO is not being produced in the dark), the reaction of RuBisCO responds positively to increasing carbon dioxide concentration, the concentration of carbon dioxide is the only limiting factor to the efficiency of the Calvin cycle. Carbon fixation by RuBisCO can be enhanced by increasing the carbon dioxide level in the compartment containing RuBisCO. Several times during the evolution of plants, mechanisms have evolved for increasing the level of carbon dioxide. Photosynthesis The primary source of energy for nearly all life is the Sun. The energy in sunlight is introduced into the biosphere by a process known as photosynthesis, which occurs in plants, some types of bacteria and algae.

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Algae are microscopic organisms that can grow via photosynthesis, grow quite rapidly, and are considerably more productive than land plants and macroalgae. Microalgae reproduction occurs primarily by vegetative (asexual) cell division, although sexual reproduction can occur in many species under appropriate growth conditions. There are several main groups of microalgae, which differ primarily in pigment composition, biochemical constituents, ultra structure, and life cycle. The specific Algaenius strain of algae is a Green Algae. Green algae, often referred to as chlorophytes, are also abundant; approximately 8,000 species are estimated to be in existence. This group has chlorophyll a and chlorophyll b. Green algae are the evolutionary progenitors of higher plants. The growth requirements of algae are very simple; minimal light, carbon dioxide and water, although the growth rates can be accelerated by sufficient aeration and the addition of nutrients. Photosynthesis can be defined as the physico-chemical process by which photosynthetic organisms use light energy to drive the synthesis of organic compounds. The photosynthetic process depends on a set of complex protein molecules that are located in and around a highly organized membrane. Through a series of energy transducing reactions, the photosynthetic machinery transforms light energy into a stable form that can last for hundreds of millions of years. In plants, algae and certain types of bacteria, the photosynthetic process results in the release of molecular oxygen and the removal of carbon dioxide from the atmosphere that is used to synthesize carbohydrates (oxygenic photosynthesis). Photosynthesis provides the energy and reduced carbon required for the survival of virtually all life on our planet, as well as the molecular oxygen necessary for the survival of oxygen consuming organisms. Although photosynthesis occurs in cells or organelles that are typically only a few microns across, the process has a profound impact on the earth's atmosphere and climate. Each year more than 10% of the total atmospheric carbon dioxide is reduced to carbohydrate by photosynthetic organisms. Most, if not all, of the reduced carbon is returned to the atmosphere as carbon dioxide by microbial, plant and animal metabolism, and by biomass combustion. The overall equation for photosynthesis is deceptively simple; a set of physical and chemical reactions (see below) occur in a coordinated manner for the synthesis of carbohydrates.

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The key feature of non-CA plant photosynthesis is that plants can use photons to make carbohydrates from CO2 and water. The empirical equation representing the net reaction of non-CA photosynthesis for oxygen evolving organisms is: CO2 + 2H2O + Light Energy [CH2O] + O2 + H2O, [CH2O] represents a carbohydrate i.e glucose, a six-carbon sugar The synthesis of carbohydrate from carbon and water requires an input of light energy. The standard non-CA free energy for the reduction of one mole of CO2 to the level of glucose is + 478 kJ/mol. Because glucose, a six-carbon sugar, is often an intermediate product of photosynthesis, the net equation of photosynthesis: 6CO2 + 12H2O + Light Energy C6H12O6 + 6O2 + 6H2O. The standard free energy for the synthesis of glucose is + 2,870 kJ/mol. The biochemical conversion of CO2 to carbohydrate is a reduction reaction that involves the rearrangement of covalent bonds between carbon, hydrogen and oxygen. The energy for the reduction of carbon is provided by energy rich molecules that are produced by the light driven electron transfer reactions. The photosynthetic process in all plants and algae (with and without CA) as well as in certain types of photosynthetic bacteria involves the reduction of CO2 to carbohydrate and removal of electrons from H20, which results in the release of O2. In this process, known as oxygenic photosynthesis, water is oxidized by the photo-system II (PS-II) reaction center, a multi-subunit protein located in the photosynthetic membrane. The photosynthetic process in plants and algae occurs in small organelles known as chloroplasts that are located inside cells. The more primitive photosynthetic
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organisms, for example oxygenic cyanobacteria, prochlorophytes and an oxygenic photosynthetic bacteria, lack organelles. The photosynthetic reactions are traditionally divided into two stages the "light reactions," which consist of electron and proton transfer reactions and the "dark reactions," which consist of the biosynthesis of carbohydrates from CO2. The light reactions occur in a complex membrane system (the photosynthetic membrane) that is made up of protein complexes, electron carriers, and lipid molecules. The photosynthetic membrane is surrounded by water and can be thought of as a two-dimensional surface that defines a closed space, with an inner and outer water phase. A molecule or ion must pass through the photosynthetic membrane to go from the inner space to the outer space. The protein complexes embedded in the photosynthetic membrane have a unique orientation with respect to the inner and outer phase. The asymmetrical arrangement of the protein complexes allows some of the energy released during electron transport to create an electrochemical gradient of protons across the photosynthetic membrane. Photosynthetic electron transport consists of a series of individual electron transfer steps from one electron carrier to another. The electron carriers are metal ion complexes and aromatic groups. The metal ion complexes and most of the aromatic groups are bound within proteins. Most of the proteins involved in photosynthetic electron transport are composed of numerous polypeptide chains that lace through the membrane, providing scaffolding for metal ions and aromatic groups. An electron enters a protein complex at a specific site, is transferred within the protein from one carrier to another, and exits the protein at a different site. The protein controls the pathway of electrons between the carriers by determining the location and environment of the metal ion complexes and aromatic groups. By setting the distance between electron carriers and controlling the electronic environment surrounding a metal ion complex or aromatic group, the protein controls pair wise electron transfer reactions. Between proteins, electron transfer is controlled by distance and free energy, as for intra-protein transfer, and by the probability that the two proteins are in close contact. Protein association is controlled by a number of factors, including the structure of the two proteins, their surface electrical and chemical properties and the probability that they collide with one another. Not all electron carriers are bound to proteins. The reduced forms of plastoquinone or ubiquinone and nicotinamide adenine dinucleotide phosphate (NADPH) or NADH act as mobile electron carriers operating between protein complexes. For electron transfer to occur, these small molecules must bind to special pockets in the proteins known as binding sites. The binding sites are highly specific and are a critical factor in controlling the rate and pathway of electron transfer.
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In oxygenic photosynthetic organisms, two different reaction centers, known as photo-system II (PS-II) and photo-system I (PS-I), work concurrently but in series. In the light photo-system II feeds electrons to photo-system I. The electrons are transferred from photo-system II (PS-II) to the photo-system I (PS-I) by intermediate carriers. The net reaction is the transfer of electrons from a water molecule to NADP+, producing the reduced form NADPH. In the photosynthetic process, much of the energy initially provided by light energy is stored as redox free energy (a form of chemical free energy) in NADPH, to be used later in the reduction of carbon if CA is not available. In addition, the electron transfer reactions concentrate protons inside the membrane vesicle and create an electric field across the photosynthetic membrane. In this process the electron transfer reactions convert redox free energy (either from the photons or the CA conversion) into an electrochemical potential of protons. The energy stored in the proton electrochemical potential is used by a membrane bound protein complex (ATP-Synthase) to covalently attach a phosphate group to adenosine diphosphate (ADP), forming adenosine triphosphate (ATP). Protons pass through the ATP-Synthase protein complex that transforms electrochemical free energy into a type of chemical free energy known as phosphate group-transfer potential (or a high-energy phosphate bond). The energy stored in ATP can be transferred to another molecule by transferring the phosphate group. The aromatics include quinones, pheophytin, NADPH, tyrosine and a flavoprotein. The NADPH and ATP formed by the reactions provide the energy for the dark reactions of photosynthesis, known as the Calvin cycle or the photosynthetic carbon reduction cycle. The reduction of atmospheric CO2 to carbohydrate occurs in the aqueous phase of the chloroplast and involves a series of enzymatic reactions. The first step is catalyzed by the protein Rubisco (D-ribulose 1, 5bisphosphate carboxylase/oxygenase), which attaches CO2 to a five-carbon compound. The reaction produces two molecules of a three-carbon compound. Subsequent biochemical reactions involve several enzymes that reduce carbon by hydrogen transfer and rearrange the carbon compounds to synthesize carbohydrates. The carbon reduction cycle involves the transfer and rearrangement of chemical bond energy. Synthesis of Carbohydrates All plants and algae remove CO2 from the environment and reduce it to carbohydrate by the Calvin cycle. The process is a sequence of biochemical reactions that reduce carbon and rearrange bonds to produce carbohydrate from CO2 molecules. The first step is the addition of CO2 to a five-carbon compound (ribulose 1,5-bisphosphate). The six-carbon compound is split, giving two molecules of a three-carbon compound (3-phosphoglycerate).

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The main energy input in the Calvin cycle is the phosphorylation by ATP and subsequent reduction by NADPH of the initial three-carbon compound forming a three-carbon sugar, triosephosphate. Some of the triosephosphate is exported from the chloroplast and provides the building block for synthesizing more complex molecules. In a process known as regeneration, the Calvin cycle uses some of the triosephosphate molecules to synthesize the energy rich ribulose 1,5bisphosphate needed for the initial carboxylation reaction. This reaction requires the input of energy in the form of one ATP. Overall, thirteen enzymes are required to catalyze the reactions in the Calvin cycle. The energy conversion efficiency of the Calvin cycle is approximately 90%. The reactions do not involve energy transduction, but rather the rearrangement of chemical energy. Each molecule of CO2 reduced to a sugar [CH2O] requires 2 molecules of NADPH and 3 molecules of ATP. Rubisco is a bi-functional enzyme that, in addition to binding CO2 to ribulose bisphosphate, can also bind O2. This oxygenation reaction produces the 3phosphoglycerate that is used in the Calvin cycle and a two-carbon compound (2phosphoglycolate) that is not useful for the plant. In response, a complicated set of reactions (known as photorespiration) are initiated that serve to recover reduced carbon and to remove phosphoglycolate. Photosynthetic Yield and Energy Conversion Efficiency The theoretical minimum quantum requirement for basic photosynthesis not including any efficiencies gained through Carbonic Anhydrase, is 8 quanta for each molecule of oxygen evolved (four quanta required by photo-system II and four by photo-system I). Measurements in algal cells under optimal conditions (e.g., low light) give quantum requirements of 8-10 photons per oxygen molecule released. These quantum yield measurements show that the quantum yields of photosystem II and photo-system I reaction centers under optimal conditions are near 100%. These values can be used to calculate the theoretical energy conversion efficiency of photosynthesis (free energy stored as carbohydrate/light energy absorbed). If 8 red quanta are absorbed (8 mol of red photons are equivalent to 1,400 kJ) for each CO2 molecule reduced (480 kJ/mol), the theoretical maximum energy efficiency for carbon reduction is 34%. Under optimal conditions, plants can achieve energy conversion efficiencies within 90% of the theoretical maximum. However, under normal growing conditions the actual performance of the plant is far below these theoretical values. The factors that conspire to lower the quantum yield of photosynthesis include limitations imposed by biochemical reactions in the plant and environmental conditions that limit photosynthetic performance (energy efficiency) to an average of 27% for generic Green Algae without the Carbonic Anhydrase Enzyme. Conclusion Since the CA enzyme assists in the rapid inter-conversion of carbon dioxide and water into carbonic acid, protons, and bicarbonate ions. The additional protons are
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liberated during this reaction, even more CO2 can be metabolized. This rate of metabolization normally increases the efficiency of a photosynthetic system by up to 300 times more efficient in terms of productivity.

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Algae as animal feed A large number of nutritional and toxicological evaluations demonstrated the suitability of algae biomass as a valuable feed supplement or substitute for conventional protein sources (soybean meal, fish meal, rice bran, etc.).About five decades ago, the mass production of certain protein-rich micro-algae was considered as a possibility to close the predicted so called protein gap. Comprehensive analyses and nutritional studies (see below) have demonstrated that these algal proteins are of high quality and comparable to conventional vegetable proteins. However, due to high production costs as well as technical difficulties to incorporate the algal material into palatable food preparations, the propagation of algal protein is still in its infancy. To date, the majority of micro-algal preparations are marketed as health food, as cosmetics or as animal feed.

However, due to high production costs as well as technical difficulties to incorporate the algal material into palatable food preparations, the propagation of algal protein is still in its infancy. To date, the majority of micro-algal preparations are marketed as health food, as cosmetics or as animal feed. Since the early fifties intense efforts have been made to explore new alternate protein sources as food supplements, primarily in anticipation of a repeatedly predicted insufficient future protein supply. For these, i.e. yeasts, fungi, bacteria and micro-algae, the name Single Cell Protein (SCP) was coined to describe the protein production from biomass, originating from different microbial sources.
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Before a new food item is declared safe for human consumption, it has to undergo a series of detailed toxicological tests to prove the harmlessness of the product. This applies especially to unconventional protein sources, which is where microalgae are grouped. By considering the available information on possible toxic properties or any other adverse effects of the different algae tested so far, it can be stated that none of these algae showed any negative effect. No serious anomalies were found either in short-term or long-term feeding experiments, or in studies on acute or chronic toxicity. All tests, including human studies, failed to reveal any evidence that would restrict the utilization of properly processed algal material. All investigations conducted so far confirm that algal biomass shows promising qualities as a novel source of protein, the average quality of most of the algae examined is equal, sometimes even superior compared to conventional plant proteins. Selected data on the amino acid profile of various algae are compiled below and compared with some basic conventional food items and a reference pattern of a well-balanced protein, recommended by WHO/FAO. It can be seen that the amino acid pattern of almost all algae compares favorably with that of the reference and the other food proteins.

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The cellulosic cell wall, which represents about 10% of the algal dry matter, poses a serious problem in digesting/utilizing the algal biomass, since it is not digestible for humans and other non-ruminants. Hence, effective treatments are necessary to disrupt the cell wall to make the protein and other constituents accessible for digestive enzymes. Several authors have studied the effect of different postharvesting treatments on the digestibility of various algal species by evaluating the PER of the treated biomass, demonstrating the important role of proper processing the algal biomass.

Reference: Micro-algae as a source of protein, E.W. Becker Medical Clinic, Department II, University of Tubingen, Immunopathological Laboratory, OtfriedMller-Str-10, 72076 Tubingen, Germany. Algae as a substitute feedstock for cattle One of the major constraints to livestock production worldwide is a shortage of protein since most of the available nutrients come from the by-products of cropproduction. Straw contributes approximately 70% of the available dry matter and oil cakes, brans and green grasses are the most common supplements used. Currently, these supplements are relatively costly and may not be available under many conditions, which necessitates the search for other inexpensive sources of supplements which can correct the nutritional imbalances imposed by straw. The possibility of using unicellular algae as a cattle feed has already been thoroughly studied. In past studies, the algal culture was grown in shallow ponds with a minimal production cost of $1.25 per ton of algal suspension production. Note: dried algal cells contained 613 g crude protein and 155 g fiber per kg (DM). Except for sulphur-containing amino acids (methionine and cystine), the essential amino acid content is favorable for the nutrition of farm animals. Algae are also a rich source of carotene, vitamin C and K, and B-vitamins. However, due to their
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cellulosic cell membrane, most algae are not efficiently utilized by monogastric (simple single chambered stomach) animals. But, pregastric fermentation by microbes enables ruminants (a mammal that digests its food in two steps) to utilize cellulose very efficiently. Furthermore, as an average, it required only 12 liters of algal suspension daily to feed a single animal (consumption of algal suspension was approximately 10% of live weight). In current studies, algal feedstock appears to improve the balance of nutrients in a straw-based diet and thus increased the efficiency of conversion of feed to growth. Nitrogen deficient straw being the main source of nutrients for ruminants in most areas of the world, the introduction of algal suspension in the feeding system would certainly help economic livestock production. Many livestock farmers raise their animals absolutely on straw and concentrate with either very little or no green grasses. This system of feeding is often associated with infertility, night blindness or even total blindness or other symptoms of vitamin A- deficiency. Algae are a very rich source of carotene and algal suspension could be a potential source of vitamin A to combat such deficiencies. Bioreactors A standard bioreactor is a device or system that supports a biologically active environment that is meant to grow cells or tissues in the context of cell culture. These bioreactors may be classified as batch, fed batch or continuous. The design of a bioreactor is quite complex since it must be continually monitored to provide optimum conditions to allow the microorganisms or cells to perform their desired function with great efficiency. Through sensors, controllers and control systems, most bioreactors monitor and control the following parameters: Gas (i.e., air, oxygen, nitrogen, carbon dioxide) Flow rates Temperature pH and dissolved oxygen levels Agitation speed/circulation rate

Fouling can harm the overall sterility and efficiency of the bioreactor. Fouling is especially problematic to the systems heat exchangers. A heat exchanger is needed to maintain the bioprocess at a constant temperature. Biological fermentation is a major source of heat; therefore in most cases bioreactors need water refrigeration. They can be refrigerated with an external jacket or, for very large vessels, with internal coils. In an aerobic process, optimal oxygen transfer is perhaps the most difficult task to accomplish. Oxygen is poorly soluble in water and even less in fermentation broths and is relatively scarce in air (20%). Oxygen transfer is usually helped by agitation that is also needed to mix nutrients and to keep the fermentation homogeneous. There are however limits to the speed of agitation, due to both high power consumption and the damage to organisms due to excessive tip speed.

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In bioreactors where the goal is grow cells or tissues for experimental or therapeutic purposes, the design is significantly different from that of industrial bioreactors. Many cells and tissues, especially mammalian, must have a surface or other structural support in order to grow, and agitated environments are often destructive to these cell types and tissues. Higher organisms also need more complex growth medium. However, when cultivating algae, several factors must be considered, and different algae have different requirements. The water must be in a temperature range that will support the specific algal species being grown. Nutrients must be controlled so algae will not be "starved" and so that nutrients will not be wasted. Additionally, the light source must not be too strong or too weak. Algae are sometimes cultured in open raceway-type ponds. But since these systems are open to the elements, they are much more vulnerable to contamination by other microorganisms, such as invasive algal species or bacteria. Because of these factors and also that there are no controls over water temperature and lighting conditions, the number of species successfully cultivated in an "open-pond" system for a specific purpose such as the production of oil, is relatively limited. A variation on the basic "open-pond" system is to close it off or to cover the pond with a greenhouse. While this usually results in a smaller system, it does take care of many of the problems associated with an open system. However, it does allow for more species to be grown, it allows the species that are being grown to stay dominant, and it extends the growing season, only slightly if unheated, and if heated it can produce year round. The potential of microalgae to be cultivated for the production pharmaceuticals, chemical intermediates, animal feedstock and clean energy is tremendous. But, because no efficient large-scale photo-bioreactor systems are yet available, open cultivation ponds are used for almost all-commercial algae production. However, it is difficult to obtain high productivity in open ponds because the temperature and light intensity vary throughout the day and year. In addition, open ponds require a large surface area, and problems with contamination could easily arise. Many closed photo-bioreactors have been used or proposed for the cultivation of microalgae; the most common are vertical or horizontal tubular, helical, and inclined or horizontal thin-panel photo-bioreactors. The critical design requirement in these photo-bioreactors is to supply light efficiently by maximizing the illumination surface-to-volume ratio of the reaction. As a result, tubes are often very narrow or the panels very thin. Some of the photo-bioreactors that work well in the laboratory may not work as well when scaled up because the surface-to-volume ratio decreases, causing poor light distribution inside the reactor; or even worse, the length of the tubes maybe so long that the alga experience oxygen poisoning.
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Photo-bioreactors Algae can also be grown in a photo-bioreactor. A photo-bioreactor (PBR) is basically a bioreactor which incorporates some type of light source. While almost anything in which it would be possible to grow algae could technically be called a photo-bioreactor, the term is more commonly used to define a closed system, as opposed to an open tank, or pond. Enclosed PBRs have the following advantages over open pond production: 1. Better control of algal culture 2. Large surface-to-volume ratio 3. Better control of gas transfer 4. Reduction in evaporation of growth medium 5. More uniform temperature 6. Better protection from outside contamination 7. Higher algal cell densities are possible. Because these systems are closed, when used to cultivate algae, everything that algae need to grow, (carbon dioxide, nutrient-rich water and light), must be introduced into the system. A photo-bioreactor can be operated in batch mode but it is also possible to provide it with a stream of sterilized water with the necessary nutrients and air dosed with carbon dioxide. Excess culture overflows is then harvested. If sufficient care is not taken, continuous bioreactors often collapse very quickly but once successfully started, they can continue operating for long periods. Algaeniuss photo-bioreactor design In most algal-cultivation systems, light only penetrates the top 2 to 3 inches of the water or growing medium. This is because as the algae grow and multiply they become so dense that they block light from reaching deeper into the pond, pipes or tank. In order to have systems that are deeper or wider than 3 inches, manufactures include various methods to agitate the water in their ponds, exposing the algae below to light and keeping algae on the surface from being over-exposed. Some such schemes are: 1) Paddle wheels that can be used to circulate the water in a pond and 2) Compressed air that can be introduced into the bottom of a pond. Algaeniuss photo-bioreactor was designed and engineered to eliminate the need for energy wasting paddle wheels and the addition of costly compressed air systems though the utilization of engineered vertical photo-bioreactor tubes. In addition to permitting maximum light penetration from Algaeniuss low wattage light sticks, these engineered vertical photo-bioreactor tubes allow for maximum CO2 utilization and minimum off-gassing due to incomplete absorption from the algae. Maximum CO2 utilization is aided by the uniquely configured air-bleed tube in the photo-bioreactor to allow the algae in the vessel to combine with the gas with positive head-pressure.

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High Efficiency Light Source As with most open systems, agitation with paddle wheels and / or compressed air is one of the few means of supplying light to algae. For smaller systems, sheets of plastic or glass can be submerged into a tank / pond, providing light directly to the algae at the right concentration i.e. Glow Plates. Considering that Algae only need about 1/10th the amount of light they receive from direct sunlight and that direct sunlight is often too strong for algae, a simple low wattage light source (see above) is more than sufficient to grow algae in a large commercial system. A few of the many critical selection criteria of a light system are the focus beam / spectrum and the light wave. If the focus beam is too narrow, the algae will not benefit from the vast majority of the spectrum. Our PBR design utilizes a wraparound light source to properly illuminate the complete hive area. Another selection issue for a light system for PBR is the ideal light wavelength. Given that the algae only require a white light and most other wavelengths simply produce heat and consume energy, a simple white light in the 1-watt per gallon range will produce a sufficient amount of energy to allow proper growth in every PBR.

Agitation and gas mixing Agitation and gas mixing is very important in the production of algae in photobioreactors. The agitation and gas mixing helps to keep the cells in suspension,
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distribute the nutrients, dispense the generated heat, improve CO2 transfer, degas the photo synthetically produced O2, prevent O2 build- up, improve mass transfer between the cells and the liquid broth and facilitate the movement of cells in and out of the illuminated part of the photo-bioreactor and prevent cell sedimentation. While the growth rates of algae have a high dependency on agitation and gas mixing, the algae can also be damaged by shear stress caused by aggressive mixing or high gas velocity. Conversely, low gas velocities tend to yield larger bubbles that form relatively slowly. These large & slow forming bubbles generally bridge the entire opening of the porous membrane, which then completely disrupts the gas mixing and gas uptake from the algae. To allow proper agitation and gas mixing without forming large slow bubbles or create shear stress to the algae due to overly aggressive mixing, a serpentine, porous rope sparger (see below) is utilized to overcome the above-mentioned issues of either to low of flow or high gas velocity. The serpentine sparger design facilitates the proper formation of mini-bubbles, while overcoming the large headpressure created by the shape of the PBR (i.e. long tube)

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Control Systems The complete control system of a single tier of PBR Tubes (5 tubes per beam* 8 beams = 40 PBR) can be achieved through utilization of gravity feed system and the simple integration of two valves: 1) Nutrients in 2) Harvested algae out This control system can also manage the temperature, aeration, pH, and other factors as required. Below is an example of the control system for a single beam (5 PBR)

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Proposed PBR layout Depending on the final footprint of the building structure that is available, the proposed layout of the photo-bioreactors should be configured as follows:

The schematic on right shows a 10-structure layout (1,600 PBR total) that could be erected in an 18750 sq ft facility.

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