Biotechnology Advances 25 (2007) 294 – 306 www.elsevier.


Research review paper

Biodiesel from microalgae
Yusuf Chisti ⁎
Institute of Technology and Engineering, Massey University, Private Bag 11 222, Palmerston North, New Zealand Available online 13 February 2007

Abstract Continued use of petroleum sourced fuels is now widely recognized as unsustainable because of depleting supplies and the contribution of these fuels to the accumulation of carbon dioxide in the environment. Renewable, carbon neutral, transport fuels are necessary for environmental and economic sustainability. Biodiesel derived from oil crops is a potential renewable and carbon neutral alternative to petroleum fuels. Unfortunately, biodiesel from oil crops, waste cooking oil and animal fat cannot realistically satisfy even a small fraction of the existing demand for transport fuels. As demonstrated here, microalgae appear to be the only source of renewable biodiesel that is capable of meeting the global demand for transport fuels. Like plants, microalgae use sunlight to produce oils but they do so more efficiently than crop plants. Oil productivity of many microalgae greatly exceeds the oil productivity of the best producing oil crops. Approaches for making microalgal biodiesel economically competitive with petrodiesel are discussed. © 2007 Elsevier Inc. All rights reserved.
Keywords: Biofuels; Biodiesel; Microalgae; Photobioreactors; Raceway ponds

Contents Introduction . . . . . . . . . . . . . . . . . . . . . . Potential of microalgal biodiesel . . . . . . . . . . . Microalgal biomass production . . . . . . . . . . . . 3.1. Raceway ponds . . . . . . . . . . . . . . . . 3.2. Photobioreactors . . . . . . . . . . . . . . . 4. Comparison of raceways and tubular photobioreactors 5. Acceptability of microalgal biodiesel . . . . . . . . . 6. Economics of biodiesel production . . . . . . . . . . 7. Improving economics of microalgal biodiesel. . . . . 7.1. Biorefinery based production strategy . . . . . 7.2. Enhancing algal biology . . . . . . . . . . . . 7.3. Photobioreactor engineering . . . . . . . . . . 8. Conclusion . . . . . . . . . . . . . . . . . . . . . . References . . . . . . . . . . . . . . . . . . . . . . . . . 1. 2. 3. . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . . 295 296 297 297 298 300 300 301 302 302 302 303 304 304

⁎ Tel.: +64 6 350 5934; fax: +64 6 350 5604. E-mail address: 0734-9750/$ - see front matter © 2007 Elsevier Inc. All rights reserved. doi:10.1016/j.biotechadv.2007.02.001

Gavrilescu and Chisti. Shimizu. as methanol boils off at 65 °C at atmospheric pressure.. 2001). Van Gerpen.. A higher temperature can be used in combination with higher pressure. hence the reaction mixture contains two liquid phases. i. Melis. 1995. In addition.. Biodiesel is a proven fuel. 2005). Under these conditions. 2003. 1997. 1991. but not from microalgae.. biodiesel is produced mainly from soybeans. Technology for producing and using biodiesel has been known for more than 50 years (Knothe et al.. reaction takes about 90 min to complete. but is not currently feasible because of the relatively high cost of the catalyst (Fukuda et al. Box 1 Biodiesel production Parent oil used in making biodiesel consists of triglycerides (Fig. 2002. Felizardo et al. Meher et al. the emerging concern about global warming that is associated with burning fossil fuels (Gavrilescu and Chisti. Alkoxides such as sodium methoxide are even better catalysts than sodium hydroxide and are being increasingly used. 2006.. Transesterification requires 3 mol of alcohol for each mole of triglyceride to produce 1 mol of glycerol and 3 mol of methyl esters (Fig. 1980–81.e. more significantly. The reaction is an equilibrium.. Sawayama et al. 2004. 2006). B1). These include methane produced by anaerobic digestion of the algal biomass (Spolaore et al. 2001.. Borowitzka. The typically used process for commercial production of biodiesel is explained in Box 1... Kapdan and Kargi. Alkalicatalyzed transesterification is carried out at approximately 60 °C under atmospheric pressure.. In the United States. The idea of using microalgae as a source of fuel is not new (Chisti. Ghirardi et al. This article focuses on microalgae as a potential source of biodiesel. Transesterification of oil to biodiesel. Kulkarni and Dalai... Spolaore et al. Singh et al.. Banerjee et al. Barnwal and Sharma. Transestrification produces methyl esters of fatty acids. Banerjee et al. 2002. Production of methyl esters. 2005. 2001). but this is expensive. B1) in which three fatty acid molecules are esterified with a molecule of glycerol. feeds and high-value bioactives (Metting and Pyne.. 2005). Lorenz and Cysewski. Kulkarni and Dalai. waste cooking oil (Felizardo et al... 2002... 1999. Dunahay et al. 2003. from microalgal oil has been demonstrated (Belarbi et al. 2005). In making biodiesel. 2001). 2002. 1995). Yield of methyl esters exceeds 98% on a weight basis (Fukuda et al.. foods.. Other alcohols can be used. alkalis (Fukuda et al. towards biodiesel. Kalin et al. Meher et al. 2006). 2006) and lipase enzymes (Sharma et al. Suresh and Ravishankar.. Melis. 2005. 1986. Other sources of commercial biodiesel include canola oil. 2001). Introduction Microalgae are sunlight-driven cell factories that convert carbon dioxide to potential biofuels. The reaction occurs stepwise: triglycerides are first converted to diglycerides. 2000. corn oil. Akkerman et al. 2002. 1998. 2002. 2005. This is likely to change as several companies are attempting to commercialize microalgal biodiesel. Akkerman et al. Fig. Alkali-catalyzed transesterification is about 4000 times faster than the acid catalyzed reaction (Fukuda et al. B1... 2006. Any future production of biodiesel from microalgae is expected to use the same process. Metzger and Largeau. Biodiesel is produced currently from plant and animal oils. but it is now being taken seriously because of the escalating price of petroleum and. these photosynthetic microorganisms are useful in bioremediation applications (Mallick. Fedorov et al. 2006).. animal fat. or biodiesel. Transesterification is catalyzed by acids. Kay. To prevent yield loss (continued on next page) . Walter et al. that are biodiesel. and glycerol (Fig. Fukuda et al. alkalis such as sodium and potassium hydroxide are commonly used as commercial catalysts at a concentration of about 1% by weight of oil. Methanol and oil do not mix. and jatropha oil (Barnwal and Sharma.. Chisti / Biotechnology Advances 25 (2007) 294–306 295 1. but methanol is the least expensive.. 2000. 2006).. 1990. Demirbas. 2006) and as nitrogen fixing biofertilizers Vaishampayan et al. 2005. biodiesel derived from microalgal oil (Roessler et al. 2002. palm oil. 2006. 1996. B1). Sheehan et al.. Sawayama et al. Consequently. Use of lipases offers important advantages. Industrial processes use 6 mol of methanol for each mole of triglyceride (Fukuda et al. 1990. 2005. Microalgae can provide several different types of renewable biofuels. 2005). R1–3 are hydrocarbon groups. and photobiologically produced biohydrogen (Ghirardi et al. 1994. 2001). 2001). then to monoglycerides and finally to glycerol. Munoz and Guieysse. 1996. Schwartz. This large excess of methanol ensures that the reaction is driven in the direction of methyl esters. 2006. 2005..Y. triglycerides are reacted with methanol in a reaction known as transesterification or alcoholysis.. 2001. 2005. Nagle and Lemke.

Ratledge and Wynn. 2000) although the product was intended for pharmaceutical use. Actual biodiesel yield per hectare is about 80% of the yield of the parent crop oil given in Table 1.700 Land area needed (M ha) a 1540 594 223 140 99 45 2 4. Spolaore et al.53 billion m3 of biodiesel annually at the current rate of consumption. the oil and alcohol must be dry and the oil should have a minimum of free fatty acids. The microalgal oil yields given in Table 1 are based on experimentally Table 1 Comparison of some sources of biodiesel Crop Corn Soybean Canola Jatropha Coconut Oil palm Microalgae b Microalgae c a b c Oil yield (L/ha) 172 446 1190 1892 2689 5950 136. 1993. that is the mass of oil produced per unit volume of the microalgal broth per day. meeting only half the existing U. Not all algal oils are satisfactory for making biodiesel. instead of microalgae. Biodiesel is recovered by repeated washing with water to remove glycerol and methanol. Guschina and Harwood. 2. transport fuel needs by biodiesel. 2006). depends on the algal growth rate and the oil content of the biomass. if microalgae are used to produce biodiesel. microalgae appear to be the only source of biodiesel that has the potential to completely displace fossil diesel. cropping area would be sufficient for producing algal biomass that satisfies 50% of the transport fuel needs (Table 1). Nannochloropsis sp. the cropping area needed to meet 50% of the U.5 For meeting 50% of all transport fuel needs of the United States. soap formation). Oil content in microalgae can exceed 80% by weight of dry biomass (Metting. Potential of microalgal biodiesel Replacing all the transport fuel consumed in the United States with biodiesel will require 0. Metzger and Largeau. Oil productivity. Microalgae commonly double their biomass within 24 h.S.. Unlike other oil crops. Clearly.. fodder and other products derived from crops. 30% oil (by wt) in biomass. however. but suitable oils occur commonly. Tetraselmis sueica Oil content (% dry wt) 25–75 28–32 20 16–37 23 25–33 N20 20–35 31–68 35–54 45–47 20–30 50–77 15–23 Box 1 (continued ) due to saponification reactions (i. 2005. In column 4 (Table 1) this area is expressed as a percentage of the total cropping area of the United States. microalgae grow extremely rapidly and many are exceedingly rich in oil. oil crops cannot significantly contribute to replacing petroleum derived liquid fuels in the foreseeable future. Using the average oil yield per hectare from various crops. as discussed later in this article.1 2. 70% oil (by wt) in biomass. waste cooking oil and animal fat cannot realistically satisfy this demand. 1996. Biomass doubling times during exponential growth are commonly as short as 3.e. Phaeodactylum tricornutum Schizochytrium sp. This scenario changes dramatically. usually in crop plants.5 Percent of existing US cropping area a 846 326 122 77 54 24 1. heterotrophic production is not as efficient as using photosynthetic microalgae. Microalgae with high oil productivities are desired for producing biodiesel. If oil palm. Dunaliella primolecta Isochrysis sp. This is demonstrated in Table 1. 24% of the total cropland will need to be devoted to its cultivation to meet only 50% of the transport fuel needs.296 Y. Depending on species. Neochloris oleoabundans Nitzschia sp. Using microalgae to produce biodiesel will not compromise production of food. This is because the renewable organic carbon sources required for growing heterotrophic microorganisms are produced ultimately by photosynthesis.5 h. Between 1 and 3% of the total U. transport fuel needs is calculated in column 3 (Table 1).S. In view of Table 1. 2006).S. Monallanthus salina Nannochloris sp. Potentially. oil producing heterotrophic microorganisms (Ratledge. 2002. Crypthecodinium cohnii Cylindrotheca sp. hydrocarbons and other complex oils (Banerjee et al. microalgae produce many different kinds of lipids. . For example. Oil levels of 20–50% are quite common (Table 2). Chisti / Biotechnology Advances 25 (2007) 294–306 Table 2 Oil content of some microalgae Microalga Botryococcus braunii Chlorella sp. would require unsustainably large cultivation areas for major oil crops.900 58. 2002) grown on a natural organic carbon source such as sugar. can be used to make biodiesel. demonstrated biomass productivity in photobioreactors. Oil crops. a high-yielding oil crop can be grown.

All of this carbon is typically derived from carbon dioxide. Raceway channels are built in concrete. In this method of operation. Flow is guided around bends by baffles placed in the flow channel. 3. 1). 1999) and tubular photobioreactors (Molina Grima et al.83N0.000 m2 (Spolaore et al. 1999)... 1999). Chisti / Biotechnology Advances 25 (2007) 294–306 297 Production of algal oils requires an ability to inexpensively produce large quantities of oil-rich microalgal biomass.11P0.. and the temperature at night. 1995. Feeding controlled in response to signals from pH sensors minimizes loss of carbon dioxide and pH variations. Sea water supplemented with commercial nitrate and phosphate fertilizers and a few other micronutrients is commonly used for growing marine microalgae (Molina Grima et al. Producing 100 t of algal biomass fixes roughly 183 t of carbon dioxide. Raceway ponds for mass culture of microalgae have been used since the 1950s. 1999). a constant rate and the same quantity of microalgal broth is withdrawn continuously (Molina Grima et al. 1997). that is CO0. Arial view of a raceway pond. Tredici. This carbon dioxide is often available at little or no cost. The largest raceway-based biomass production facility occupies an area of 440. phosphorus (P). Carbon dioxide must be fed continually during daylight hours.. carbon dioxide.1. Minimal nutritional requirements can be estimated using the approximate molecular formula of the microalgal biomass. may be lost during the night because of respiration. The only practicable methods of large-scale production of microalgae are raceway ponds (Terry and Raymond. 3. as all the power needed for producing and processing the algae would come from biodiesel itself and from methane produced by anaerobic digestion of biomass residue left behind after the oils has been extracted. despite daily and seasonal variations in light levels.48H1. Extensive experience exists on operation and engineering of raceways. but the mixing of broth must continue to prevent settling of the biomass (Molina Grima et al.. Raceway ponds A raceway pond is made of a closed loop recirculation channel that is typically about 0. 2006). 1999. To minimize expense. and may be lined with white plastic. Essential elements include nitrogen (N). 1). fresh culture medium is fed at Fig. Growth medium must provide the inorganic elements that constitute the algal cell. Microalgal biomass contains approximately 50% carbon by dry weight (Sánchez Mirón et al.. During daylight. 1999).... Sánchez Mirón et al. water and inorganic salts. Microalgal biomass production Producing microalgal biomass is generally more expensive than growing crops. The extent of this loss depends on the light level under which the biomass was grown. Photosynthetic growth requires light. Broth is harvested behind the paddlewheel. Although microalgal biodiesel can be carbon neutral. Growth media are generally inexpensive. on completion of the circulation loop. 1985. Yun et al. As much as 25% of the biomass produced during daylight.3 m deep (Fig. Temperature must remain generally within 20 to 30 °C. This facility. as discussed next. This formula is based on data presented by Grobbelaar (2004). Feeding ceases during the night.Y. 1.01. biodiesel production must rely on freely available sunlight. 1999. therefore. the culture is fed continuously in front of the paddlewheel where the flow begins (Fig. it will not result in any net reduction in carbon dioxide that is accumulating as a consequence of burning of fossil fuels. iron and in some cases silicon. . the growth temperature. not all the added P is bioavailable. 2003). Mixing and circulation are produced by a paddlewheel (Fig. 1). Molina Grima. Nutrients such as phosphorus must be supplied in significant excess because the phosphates added complex with metal ions. The paddlewheel operates all the time to prevent sedimentation. Large-scale production of microalgal biomass generally uses continuous culture during daylight. Ideally. or compacted earth. Biodiesel production can potentially use some of the carbon dioxide that is released in power plants by burning fossil fuels (Sawayama et al. microalgal biodiesel would be carbon neutral.

The ground beneath the solar collector is often painted white. 2). 2). Fig. 2. 1999. and back to the reservoir. Because of significant losses to atmosphere. 2001. Chisti / Biotechnology Advances 25 (2007) 294–306 owned by Earthrise Nutritionals (www. Although raceways are low-cost. Microalgal broth is circulated from a reservoir (i. The tubes are always oriented North–South (Fig.. 2006). 1999). 2) to the solar collector Fig.. The solar collector is oriented to maximize sunlight capture (Molina Grima et al.1 m or less in diameter. 3). any cooling is achieved only by evaporation. 3.298 Y. 1999.2.. Carvalho et Continuous culture operation is used. 1999. as explained above. Photobioreactors have been successfully used for producing large quantities of microalgal biomass (Molina Grima et al. raceways use carbon dioxide much less efficiently than photobioreactors. Tube diameter is limited because light does not penetrate too deeply in the dense culture broth that is necessary for ensuring a high biomass productivity of the photobioreactor. Pulz. In a typical arrangement. Temperature fluctuates within a diurnal cycle and seasonally. Sánchez Mirón et al. Evaporative water loss can be significant. A fence-like solar collector. is where the sunlight is captured (Fig. Productivity is affected by contamination with unwanted algae and microorganisms that feed on algae. A tubular photobioreactor consists of an array of straight transparent tubes that are usually made of plastic or glass. Photobioreactors Unlike open raceways. A tubular photobioreactor with parallel run horizontal tubes. Courtesy of Professor Michael Borowitzka. because they cost less to build and operate. Australia. 3. A 1000 L helical tubular photobioreactor at Murdoch University. the degassing column in Fig. Murdoch University. in attempts to increase the number of tubes that can be accommodated in a given area. Tredici. they have a low biomass productivity compared with photobioreactors. Raceways are perceived to be less expensive than photobioreactors. Production of microalgal biomass for making biodiesel has been extensively evaluated in raceway ponds in studies sponsored by the United States Department of Energy (Sheehan et al. The solar collector tubes are generally 0.. This tubular array. the solar tubes are placed parallel to each other and flat above the ground (Fig. or covered with white sheets of plastic Fig. In raceways. 1998). Raceway ponds and other open culture systems for producing microalgae are further discussed by Terry and Raymond (1985). 3). . is used to produce cyanobacterial biomass for food. The biomass concentration remains low because raceways are poorly mixed and cannot sustain an optically dark zone. photobioreactors permit essentially single-species culture of microalgae for prolonged durations..earthrise. 4. parallel straight tubes are sometimes arranged like a fence (Fig.e. Horizontal. or the solar collector.

or a gentler airlift pump. 12 m wide.384 5681 136. a high concentration of dissolved oxygen in combination with intense sunlight produces photooxidative damage to algal cells. Based on 70% by wt oil in biomass. Chisti. Other variants of tubular photobioreactors exist (Molina Grima et al. 2001. 1993. Carvalho et al. Chisti. 2001. The culture must periodi- cally return to a degassing zone (Fig.. 2) that are needed for producing biodiesel. and the concentration of oxygen at the entrance of tube. 2001).035 b 0.00 0.. Pulz. 2000.333 132 parallel tubes/unit. Gas–liquid separator design for achieving complete disengagement of bubbles..048 a 0. 4).. photobioreactors must be cleaned and sanitized. Based on actual pond area. the degassing zone (Fig. Carbon dioxide is fed in the degassing zone in response to a pH controller.. Mechanical pumps can damage the biomass (Chisti. Acién Fernández et al. Periodically.. 2001). the flow rate. Flow is produced using either a mechanical pump (Fig. 2) must disengage all the gas bubbles from the broth so that essentially bubblefree broth returns to the solar collector tubes. 2003.. Dissolved oxygen levels much greater than the air saturation values inhibit photosynthesis (Molina Grima et al. 2).6 e 183. Chisti and Moo-Young. 1999). artificial illumination has been used in large-scale biomass production (Pulz. 1994. García Camacho et al. 2006). but expensive compared with natural illumination. the maximum tolerable dissolved oxygen level should not generally exceed about 400% of air saturation value. for inoculating the larger tubular photobioreactors (Fig. install and operate. or albedo. Additional carbon dioxide injection points may be necessary at intervals along the tubes.14 0. Airlift pumps for use in tubular photobioreactors are designed using the same methods that were originally developed for designing conventional airlift reactors (Chisti et al. Based on 30% by wt oil in biomass.. Table 3 Comparison of photobioreactor and raceway production methods Variable Annual biomass production (kg) Volumetric productivity (kg m− 3 d− 1) Areal productivity (kg m− 2 d− 1) Biomass concentration in broth (kg m− 3) Dilution rate (d− 1) Area needed (m2) Oil yield (m3 ha− 1) Annual CO2 consumption (kg) System geometry Photobioreactor facility 100. 4 are potentially useful for growing a small volume of microalgal broth. To prevent inhibition and damage.535 0. . Photobioreactors such as the one shown in Fig. but are easy to design. 2001) particularly for high-value products. 2001. it is poorly illuminated and. to prevent carbon limitation and an excessive rise in pH (Molina Grima et al. 1999). Because a degassing zone is generally optically deep compared with the solar collector tubes.000 0. 1988. Typically. to increase reflectance. As the broth moves along a photobioreactor tube. 1998).30 m deep 8 Number of units a b c d e Based on facility area. 2) that is bubbled with air to strip out the accumulated oxygen. 1999.250 7828 99. 82 m long..000 1.Y. but are not widely used. Mazzuca Sobczuk et al.. 1999a. 1999). the maximum rate of oxygen generation in a typical tubular photobioreactor may be as high as 10 g O2 m− 3 min− 1. Furthermore. 2001).9 d 58.. therefore. 1999b). Photosynthesis generates oxygen. 1989)..333 978 m2/pond. Chisti. Airlift pumps have been used quite successfully (Molina Grima et al. A high albedo increases the total light received by the tubes. 1993.06 m tube diameter 6 Raceway ponds 100. the tubes may be made of flexible plastic and coiled around a supporting frame to form a helical coil tubular photobioreactors (Fig. This is easily achieved using automated clean-inplace operations (Chisti and Moo-Young.4 d 42. Chisti / Biotechnology Advances 25 (2007) 294–306 299 (Tredici. a continuous tube run should not exceed 80 m (Molina Grima et al. Oxygen cannot be removed within a photobioreactor tube. the light intensity. but the exact length depends on several factors including the concentration of the biomass. has been discussed (Chisti and Moo-Young. In addition to removing the accumulated dissolved oxygen. its volume needs to be kept small relative to the volume of the solar collector.117 0. 2006). Based on projected area of photobioreactor tubes. pH increases because of consumption of carbon dioxide (Camacho Rubio et al. 2007. Tredici. 0. This limits the maximum length of a continuous run tube before oxygen removal becomes necessary. Under high irradiance. 2001). Instead of being laid horizontally on the ground. for example. 0. 1988. 80 m long tubes. Artificial illumination of tubular photobioreactors is technically feasible (Pulz. Biomass sedimentation in tubes is prevented by maintaining highly turbulent flow. Nonetheless. 1999. Airlift pumps are less flexible than mechanical pumps and require a supply of air to operate.7 e 183. Sánchez Mirón et al. 1999.072 c 4.

Thus. centrifugation. Microalgal biomass recovered from the culture broth by filtration moves along a conveyor belt at Cyanotech Corporation (www. For example. 2001. 3-double bonds). In the United States the relevant standard is the ASTM Biodiesel Standard D 6751 (Knothe. Furthermore. . eicosapentaenoic acid (EPA. Some vegetable oils also face this problem. Spolaore et al. Courtesy of Honolulu Star-Bulletin. 1999). six double bonds) occur commonly in algal oils. Outdoor tubular photobioreactors are effectively and inexpensively cooled using heat exchangers. separate standards exist for biodiesel intended for vehicle use (Standard EN 14214) and for use as heating oil (Standard EN 14213) (Knothe.. USA. the European Standard EN 14214 limits linolenic acid methyl ester content in biodiesel for vehicle use to 12% (mol). 2006). This is because the typical biomass concentration that is produced in photobioreactors is nearly 30 times the biomass concentration that is generally obtained in raceways (Table 3). Molina Grima et al. 2001). can also be used and has proven successful in dry climates. Evaporative cooling by water sprayed on tubes (Tredici. Molina Grima. For example. The production methods in Table 3 are compared for optimal combinations of biomass productivity and concentration that have been actually achieved in large-scale photobioreactors and raceways. No such limitation exists for biodiesel intended for use as heating oil. the nightly loss of biomass due to respiration can be reduced by lowering the temperature at night. For example. Microalgal oils differ from most vegetable oils in being quite rich in polyunsaturated fatty acids with four or more double bonds (Belarbi et al. Photobioreactors provide much greater oil yield per hectare compared with raceway ponds (Table 3). 5. if losses to atmosphere are disregarded. C20:5n-3. 2000). 2-double bonds) and linolenic acid (C18:3n-3.. Biomass is easily recovered from the broth by filtration (Fig. five double bonds) and docosahexaenoic acid (DHA. Alternatively. Hawaii. Fatty acids and fatty acid methyl esters (FAME) with 4 and more double bonds are susceptible to oxidation during storage and this reduces their acceptability for use in biodiesel. Chisti / Biotechnology Advances 25 (2007) 294–306 Photobioreactors require cooling during daylight hours. Both raceway and photobioreactor production methods are technically feasible.. Tredici. Lorenz and Cysewski.. 2006). 2003). Biomass recovery from photobioreactor cultured broth costs only a fraction of the recovery cost for broth produced in raceways. 1999. Recovery of microalgal biomass from the broth is necessary for extracting the oil. C22:6n-3. 1999. Although these fatty acids have much higher oxidative stability compared with DHA and EPA. Comparison of raceways and tubular photobioreactors Table 3 compares photobioreactor and raceway methods of producing microalgal biomass. 2003. 2006). Large tubular photobioreactors have been placed within temperature controlled greenhouses (Pulz. Acceptability of microalgal biodiesel For user acceptance. but doing so is prohibitively expensive for producing biodiesel. 2). In European Union. vegetable oils such as high oleic canola oil contain large quantities of linoleic acid (C18:2n-6. 1985. in comparison with raceway broth. heat exchangers may be placed in the tubular loop. and other means (Molina Grima et al. temperature control at night is also useful. Cost of biomass recovery can be significant. 5). 1999. Photograph by Terry Luke. A heat exchange coil may be located in the degassing column (Fig. 5. Pulz. but Fig.cyanotech. This is because the volumetric biomass productivity of photobioreactors is more than 13-fold greater in comparison with raceway ponds (Table 3). Both production methods consume an identical amount of carbon dioxide (Table 3). microalgal biodiesel will need to comply with existing standards. Selecting a suitable microalgal biomass production method for making biodiesel requires a comparison of capabilities of raceways and tubular photobioreactors. much smaller volume of the photobioreactor broth needs to be processed to obtain a given quantity of biomass. This comparison is for an annual production level of 100 t of biomass in both cases.300 4. Production facilities using photobioreactors and raceway units of dimensions similar to those in Table 3 have indeed been used extensively in commercial operations (Terry and Raymond.

as follows: Calgal oil ¼ 6:9 Â 10−3 Cpetroleum ð1Þ where Calgal oil ($ per liter) is the price of microalgal oil and Cpetroleum is the price of crude oil in $ per barrel. then microalgal oil costing $0. if it is to substitute for crude oil. because of economy of scale. This suggests that the process of converting palm oil to biodiesel adds about $0. The estimation methods used have been described previously (Humphreys. For the facilities detailed in Table 3.81 for photobioreactors and raceways.66 and $0. Using the same analogy. Total unsaturation of an oil is indicated by its iodine value. These estimates assume that carbon dioxide is available at no cost. Chisti / Biotechnology Advances 25 (2007) 294–306 301 acceptable biodiesel must meet other criteria relating to the extent of total unsaturation of the oil. Oil recovered from the lower-cost biomass produced in photobioreactors is estimated to cost $2. cost of crude oil (52%).Y.60 for photobioreactors and raceways. sold for an average price of $465/t.. 1991.66/L.. Elimination of dependence on petroleum diesel and environmental sustainability require reducing the cost of production of algal oil from about $2.40 and $1.72.49/L with a 73% contribution from crude oil and 27% contribution from refining. In the United States during 2006. 6. 6.14/L to the price of oil. 2003). Furthermore. but this need not be a significant limitation. distribution and marketing (9%). a reasonable target price for microalgal oil is $0. Biodiesel from palm oil costs roughly $0. respectively. the cost of producing the biomass is the only relevant factor for a comparative assessment of photobioreactors and raceways for producing microalgal biodiesel. respectively.48/L. Molina Grima et al. Microalgal biodiesel refinery: producing multiple products from algal biomass. This assumes that the recovery process contributes 50% to the cost of the final recovered oil. or about $0. Economics of biodiesel production Recovery of oil from microalgal biomass and conversion of oil to biodiesel are not affected by whether the biomass is produced in raceways or photobioreactors. This price included taxes (20%). Eq. the on-highway petrodiesel price ranged between $0. that is probably the cheapest vegetable oil. the average price of petrodiesel in 2006 was $0. crude palm oil. Hence. to a maximum of 1 % mol. This is a strategic objective. These desired levels of cost reduction are substantial. microalgal oil will need to be sourced at a price that is roughly related to the price of crude oil. For palm oil sourced biodiesel to be competitive with petrodiesel. For example. respectively. Dijkstra. Assuming that the biomass contains 30% oil by weight. or 35% more than petrodiesel. In view of the composition of many microalgal oils. refining expenses (19%). both the European biodiesel standards limit the contents of FAME with four and more double bonds.55/L is likely to economically substitute for crude petroleum. For this to happen. If the annual biomass production capacity is increased to 10. then microalgal oil should not cost more than about $0. The extent of unsaturation of microalgal oil and its content of fatty acids with more than 4 double bonds can be reduced easily by partial catalytic hydrogenation of the oil (Jang et al.000 t. the estimated cost of producing a kilogram of microalgal biomass is $2. The cost reduction necessary declines to $0. the price of palm oil should not exceed $0. the cost of biomass for providing a liter of oil would be something like $1.48/L for algal diesel to be cost competitive with petrodiesel.95 and $3.52/L. Microalgal oils can potentially completely replace petroleum as a source of hydrocarbon feedstock for the petrochemical industry.47 and $0. Fig. respectively.80/L to $0. assuming an absence of tax on biodiesel.48/ L. 2005. if the algal biomass is produced in photobioreactors and contains 70% oil by weight. (1) assumes that algal oil has roughly 80% of the energy content of crude petroleum. if the prevailing price of crude oil is $60/barrel. but attainable. 2006).79/L. the same technology that is commonly used in making margarine from vegetable oils. If the price of crude oil rises to $80/barrel as sometimes predicted.41/L.80/L. In comparison with this. the cost of production per kilogram reduces to roughly $0.80 for photobioreactors and raceways. most of them are unlikely to comply with the European biodiesel standards. . during 2006. Standards EN 14214 and EN 14213 require the iodine value of biodiesel to not exceed 120 and 130 g iodine/ 100 g biodiesel. If taxes and distribution are excluded.

Some of the residual biomass may be used to produce methane by anaerobic digestion. increase oil content in biomass. μmax. microalgal biomass contains significant quantities of proteins. Nevertheless. This approach can be used to reduce the cost of making microalgal biodiesel. and Germany for producing biofuels and other products from crops such as corn and soybean. it cannot compete with the many other low-cost organic substrates that are available for anaerobic digestion. improve temperature tolerance to reduce the expense of cooling. depending on the specific microalgae used. for generating the electrical power necessary for running the microalgal biomass production facility. carbohydrates and other nutrients (Sánchez Mirón et al. Improving economics of microalgal biodiesel Cost of producing microalgal biodiesel can be reduced substantially by using a biorefinery based production strategy.. Fig.. Because of light saturation. algal biomass residue remaining after the extraction of oil can be used potentially to make methane.1. Because all components of the biomass are used. 3. 6). 1968) and −2 s− 1 (Molina Grima et al.. the light saturation constants for microalgae Phaeodactylum tricornutum and Porphyridium cruentum are 185 μE m− 2 s− 1 (Mann and Myers. Extraction of other high-value products may be feasible. the United States. 7. improving capabilities of microalgae through genetic engineering and advances in engineering of photobioreactors. Elimination of photoinhibition or its postponement to higher light intensities can greatly increase the average daily growth rate of algal biomass. Enhancing algal biology Genetic and metabolic engineering are likely to have the greatest impact on improving the economics of production of microalgal diesel (Roessler et al. This phenomenon is known as photoinhibition. In comparison with these values. 2003). Biorefinery based production strategy Like a petroleum refinery. Microalgae become photoinhibited at light intensities only slightly greater than the light level at which the specific growth rate peaks. In addition to oils. increase photosynthetic efficiency to enable increased biomass yield on light. 4. enhance biomass growth rate.2. 2000). 1994. Light saturation constants for microalgae tend to be much lower than the maximum sunlight level that occurs at midday. Excess power could be sold to defray the cost of producing biodiesel. 2003). 2007) is technically feasible. Dunahay et al. 2004). 2000. Raven and Gregersen. B2). B2).. Genetic modification of microalgae has received little attention (León-Bañares et al.302 Y. Photoinhibition results from generally reversible damage to the photosynthetic apparatus. 2.. . Integrated biorefineries are already being operated in Canada. the residual biomass from biodiesel production processes can be used potentially as animal feed (Fig. Effect of light intensity on specific growth rate of microalgae. For example. Chisti / Biotechnology Advances 25 (2007) 294–306 7. a further increase in light level actually reduces the biomass growth rate (Fig. animal feed. Therefore. the biomass growth rate is much lower than would be possible if light saturation value could be increased substantially.. ˜ 200 μE m respectively. the typical midday outdoor light intensity in equatorial regions is about 2000 μE m− 2 s− 1. 6). Molecular level engineering can be used to potentially: 1. B2. biogas and electrical power (Fig. the overall cost of producing any given product is lowered. Above a certain value of light intensity. 1996). as a consequence of excessive light (Camacho Rubio et al. a biorefinery uses every component of the biomass raw material to produce useable products. Box 2 Light saturation and photoinhibition Light saturation is characterized by a light saturation constant (Fig.. that is the intensity of light at which the specific biomass growth rate is half its maximum value. 7. Although the use of microalgal biomass directly to produce methane by anaerobic digestion (Mata-Alvarez et al. A microalgal biorefinery can simultaneously produce biodiesel.

2003. Photobioreactor engineering Although a capability for reliable engineering and operation of tubular photobioreactors has emerged (Acién Fernández et al. 2000. 2006). 2007. 2003. enzymes that . Light intensity in the photolimited zone is lower than the saturation light level (Box 2).. The frequency of light–dark cycling depends on several factors. Janssen et al. 3. 2001.. Chisti / Biotechnology Advances 25 (2007) 294–306 303 5. 2001).. 1995. 7. 1999. Distinct from the productivity enhancing effect of light–dark cycling. microalgae are damaged by intense hydrodynamic shear fields that occur in high-velocity flow in pipes. 1990. 2001).. for example. García Camacho et al. Chisti. 2003) is difficult to achieve (Molina Grima et al. Such mixers have proved useful in other tubular reactors (Chisti et al. Robust methods for controlling wall growth are needed. problems remain. Some algae will preferentially grow attached to the internal wall of the photobioreactor tube. reduce susceptibility to photooxidation that damages cells. Camacho Rubio et al. concentration of cells.. 1989. turbulence in a dense culture reduces photoinhibition and photolimitation by ensuring that the algal cells do not reside continuously in either the well lit zone or the dark zone for long periods. light–dark cycling of above a certain frequency can increase biomass productivity relative to the case when the same quantity of light is supplied continuously over the same total exposure time (Philliphs and Myers. existing designs of motionless mixers are not satisfactory for photobioreactors because they substantially reduce penetration of light in the tubes. Camacho Rubio et al.. motionless mixers installed inside photobioreactor tubes can be used to substantially enhance the mixing between the peripheral lit zone and the interior dark zone (Molina Grima et al.. use of large slugs of air to intermittently scour the internal surface of the tube. 2000. Various attempts have been made to estimate the frequency of light–dark cycling (Molina Grima et al. Carvalho et al. Photobioreactor tubes operated with high-density culture for attaining high productivity. New designs of motionless mixers are needed. 2003). but this problem remains unresolved. 2001.. 1999a.. 1996.. there is a need to identify possible biochemical triggers and environmental factors that might favor accumulation of oil. 2001..Y. 2001. 1986. Some algae are more sensitive to shear damage than others.. 2. 1999.. Camacho Rubio et al. Thakur et al. Grobbelaar et al.3. 2001. 1999. circulation of close fitting balls in continuous run tubes to clean the internal surface. 1999). 1999a). 2006). Like cells of higher plants (Moo-Young and Chisti. 1996. including the intensity of turbulence. Under conditions of sufficient and excess external irradiance. 6. 1999. but improved methods for doing so are emerging (Sánchez Pérez et al.. 1999. thus preventing light penetration into the tube and reducing bioreactor productivity. but have been barely examined for microalgae. 1999. 2003). Intensities of shear stress are not easily determined in bioreactors (Chisti and Moo-Young. Sánchez Mirón et al.. 2001. Light–dark cycling times of 10 ms. Shear sensitivity can pose a significant problem as the intensity of turbulence needed in photobioreactors to generate optimal light–dark cycling (Grobbelaar et al. Camacho Rubio et al... Sánchez Mirón et al. Molina Grima et al.. eliminate the light saturation phenomenon (Box 2) so that growth continues to increase in response to increasing light level. 2000. are known to improve growth compared with continuous illumination of equal cumulative quantity. Terry. Potentially.. García Camacho et al. Wall growth is controlled by some of the following methods: 1.. 1994. 1998.. Janssen et al.. 2000. Methods have been developed to reduce the damage associated with turbulence of limited intensity (García Camacho et al. and 4. 2001.. 1999. Beneficial effects of rapid light–dark cycling under light saturation conditions are associated with the short dark period allowing the photosynthetic apparatus of the cells to fully recover from the excited state of the previous illumination event. Mazzuca Sobczuk et al. and the external irradiance level (Molina Grima et al. In principle. 2003. 1996).. Chisti. 1997. 2006). Richmond. 1988) and animals (Zhang et al. Mazzuca Sobczuk et al... 2001. 2005). inevitably contain a photolimited central dark zone and a relatively better lit peripheral zone (Molina Grima et al. 2000. Chisti. 1998. optical properties of the culture. suspended sand or grit particles to abrade any biomass adhering to the internal surface. 1996... Sánchez Mirón et al. 1989. 2004) without damaging algal cells. 2004). Grobbelaar. pumps and mixing tanks (Chisti. 2000.. 1953. the diameter of the tube. In addition. 2006). and 7. Nedbal et al. Unfortunately. highly turbulent flow.. Sánchez Mirón et al. reduce photoinhibition (Box 2) that actually reduces growth rate at midday light intensities that occur in temperate and tropical zones. Turbulence in the tube causes rapid cycling of the fluid between the light and dark zones. Stability of engineered strains and methods for achieving stable production in industrial microbial processes are known to be important issues (Zhang et al..

hydraulics and transport phenomena. Moo-Young M. On the calculation of shear rate and apparent viscosity in airlift and bubble column bioreactors. Chisti Y. Moo-Young M. J Chem Technol Biotechnol 1988. Eur J Lipid Sci Technol 2006. A process for high yield and scaleable recovery of high purity eicosapentaenoic acid esters from microalgae and fish oil. Chisti Y. Sonobioreactors: using ultrasound for enhanced microbial productivity. Chisti Y. Chisti Y. p. Appl Biochem Biotechnol 1996. Use of the biorefinery concept and advances in photobioreactor engineering will further lower the cost of production. Encyclopedia of bioprocess technology: fermentation. Sharma R. Bioprocess intensification approaches (Chisti and Moo-Young. Botryococcus braunii: a renewable source of hydrocarbons and other chemicals. Sánchez Mirón A. 313–52. Airlift bioreactors.18:420–32. Chisti Y. García Camacho F. Pharmaceuticals and agrochemicals from microalgae. Roessler PG. In: Cohen Z. p. 1806–15. Appl Mech Rev 1998. Jarvis EE. Biotechnol Bioeng 2003. Academic Press. Halard B. Molina Grima E. Trends Biotechnol 2000. Pneumatically agitated bioreactors in industrial and environmental bioprocessing: hydrodynamics.59:4369–76. Biotechnol Bioeng 1997. Fernández Sevilla JM. and bioseparation. Biotechnol Bioeng 1998. editor.27:1195–208.62: 71–86. Chisti Y. Chisti Y. may be used for controlling wall growth. Moo-Young M. Mass transfer in external-loop airlift bioreactors using static mixers. García Camacho F. Sánchez Pérez JA. Animal-cell damage in sparged bioreactors. Chem Eng Sci 2001. Acién Fernández FG.81:459–73.55:701–14. Sánchez Pérez JA. Wiley.26: 516–29.58:605–11. Liquid circulation in airlift reactors. Elsevier. Moo-Young M. 2379–406. Chisti Y. Sharma MP. Economics of producing microalgal biodiesel need to improve substantially to make it competitive with petrodiesel. Patel P. 8. Int J Hydrogen Energy 2002.74A:575–83. A mechanistic model of photosynthesis in microalgae. Chisti Y. microalgal biodiesel is technically feasible. Acién Fernández FG. Carvalho AP. Airlift-driven external-loop tubular photobioreactors for outdoor production of microalgae: assessment of design and performance. Conclusion As demonstrated here. Drew SW.68:45–50. Dijkstra AJ. Camacho Rubio F. Chisti / Biotechnology Advances 25 (2007) 294–306 Borowitzka MA. J Ramsay Soc 1980–81. 1999. Modern systems of plant cleaning. Chem Eng Sci 1988. Molina Grima E. Kasper M. Mixing in bubble columns: a new approach for characterizing dispersion coefficients. Dais SS. Sánchez Pérez JA. Effects of dilution rate. Renew Sustain Energy Rev 2005. It is the only renewable biodiesel that can potentially completely displace liquid fuels derived from petroleum. Microalgal reactors: a review of enclosed system designs and performances. tube diameter and solar irradiance. Chisti Y. editors. Moo-Young M. Sánchez Pérez JA. In view of their much greater productivity than raceways. Chisti Y. Chisti Y. p. Crit Rev Biotechnol 2002. Acién Fernández FG. Trends Biotechnol 2003. Chisti Y. Manipulation of microalgal lipid production using genetic engineering. Clean-in-place systems for industrial bioreactors: design. Revisiting the formation of trans isomers during partial hydrogenation of triacylglycerol oils.57–58:223–31.42:211–9. Producing low-cost microalgal biodiesel requires primarily improvements to algal biology through genetic and metabolic engineering. Biotechnol Bioeng 1989. Chem Eng Prog 1993. References Acién Fernández FG.43:451–7. Shear sensitivity. validation and operation. Rocha J. Encyclopedia of food microbiology. García Camacho F. Chisti Y. Molina Grima E. 1989. 355. Barnwal BK. Wijffels RH. Malcata FX. Chisti Y. Cerón García MC. Chisti Y. tubular photobioreactors are likely to be used in producing much of the microalgal biomass required for making biodiesel. Moo-Young M. Molina Grima E. Trans I Chem E 1996.108(3):249–64. 1999a. Can J Chem Eng 1990. A model for light distribution and average solar irradiance inside outdoor tubular photobioreactors for the microalgal mass culture. 5. 1996. Hydrodynamic damage to animal cells. An unusual hydrocarbon. Banerjee UC. Batt C. Dunahay TG. Fernández Sevilla JM. Fernández Sevilla J. but the level of improvement necessary appears to be attainable. Modelling of biomass productivity in tubular photobioreactors for microalgal cultures. Moo-Young M. Belarbi E-H.22:245–79. digest the polymer glue that binds algal cells to the tube walls. Prediction of liquid circulation velocity in airlift reactors with biological media. vol. Taylor & Francis. Molina Grima E. García Camacho F. Chem Eng Sci 2004.89(6):38–45.51:33–112.21:89–93. editors. 2003) that have proved so successful in improving the economics of various biotechnology based processes have been barely assessed for use with photobioreactors. García Camacho F. Chisti. 1999b. Chisti Y.27–28: 24–6. In: Robinson R.304 Y. Improve the performance of airlift reactors. biocatalysis.13:201–7. Molina Grima E. Fernández Sevilla JM. Janssen M. Enzyme Microb Technol 2000. Pror Energy Combust Sci 2005. Banerjee A. Molina Grima E. Photobioreactors provide a controlled environment that can be tailored to the specific demands of highly productive microalgae to attain a consistently good annual yield of oil. Prediction of dissolved oxygen and carbon dioxide concentration profiles in tubular photobioreactors for microalgal culture. . p. Meireles LA. J Ind Microbiol 1994. Prospects of biodiesel production from vegetables oils in India.9:363–78.31(5–6):466–87.21:67–110.56:2721–32. Crit Rev Biotechnol 2001. In: Flickinger MC.22:1490–506. Biotechnol Prog 2006. Bioprocess intensification through bioreactor engineering. Chisti Y. Biodiesel production from vegetable oils via catalytic and non-catalytic supercritical methanol transesterification methods. Chemicals from microalgae. Akkerman I.34:1391–2. Photobiological hydrogen production: photochemical efficiency and bioreactor design. Biotechnol Bioeng 1999. Camacho Rubio F. Chisti Y. Camacho Rubio F. Demirbas A. Chisti Y.

121124:403–12. Mace S. Microscopic green algae and cyanobacteria in high-frequency intermittent light. León-Bañares R.20:491–515. Galváan A.45:160–86. Tichý V. Biotechnological significance of toxic marine dinoflagellates. Molina Grima E. 2004.8:335–43.Y. Mann JE. Chisti Y. Waste Manag 2006. Nagle N. Richmond A. Largeau C. Melis A. Meinrath G. Grobbelaar J. Anaerobic digestion of organic solid wastes.30:555–73. Belarbi EH. et al. J Biotechnol 1999. Sánchez Mirón A. J Appl Phycol 1994. Cerón García MC. Bioresour Technol 2000.28: 243–50.40:2799–815. Chisti Y. Wijffels RH.66:486–96. Molina Grima E.23:471–99. Enzyme Microb Technol 1986.29: 602–10. Kay RA. Microalgae: a green source of renewable H2. Biotechnological potential of immobilized algae for wastewater N. Molina Grima E. Chisti Y.78:151–77. On pigments. García Camacho F. Tubular photobioreactor design for algal cultures. Biodiesel fuel production by transesterification of oils. 305 Mallick N. Nedbal L. Effects of agitation on the microalgae Phaeodactylum tricornutum and Porphyridium cruentum. Bio-hydrogen production from waste materials. Blackwell. J Biotechnol 2001. Mendes JF. 2004. Berkemeier R. Chisti Y.18:160–7. 125–77. Felizardo P. Kondo A. Munoz R. Jung MY. Gregersen KH. Analyzing biodiesel: standards and other methods. An overview of research achievements and perspectives.11:278–84. Dunn RO. Ghirardi ML. Guieysse B. Tramper J. p. The removal of uranium from mining waste water using algal/microbial biomass. ACS Symp Ser 1997. J Appl Phycol 1996.45:2901–13. Renew Sustain Energy Rev 2006. Molina Grima E. Hydrogenation for low trans and high conjugated fatty acids. Tichy V. Ghirardi ML. 1991. Knothe G. Recovery of microalgal biomass and metabolites: process options and economics. challenges and prospects. Metzger P. Water Res 2006. Commercial potential for Haematococcus microalga as a natural source of astaxanthin.51:1–51. In: Richmond A. Encyclopedia of bioprocess technology: fermentation. Biotechnology — a sustainable alternative for chemical industry. Zhang JP.25:176–94. Cysewski GR. Biotechnol Adv 2003.92: 113–31. . Fernández J. Noda H. Appl Microbiol Biotechnol 2005. Raven RPJM. Renew Sustain Energy Rev 2007. Continuous H2 photoproduction by Chlamydomonas reinhardtii using a novel two-stage. Belarbi EH. Int J Hydrogen Energy 2002.4:349–55. J Appl Phycol 2000. Biodiversity and application of microalgae. García Camacho F. Production of biodiesel from waste frying oil. García Camacho F.26:115–21.11:116–32. editor. Mur LR. Kalin M. Handbook of microalgal culture: biotechnology and applied phycology. Algal–bacterial processes for the treatment of hazardous contaminants: a review. Lipids and lipid metabolism in eukaryotic algae. Grobbelaar JU. Wheeler WN. Chile T. Appl Biochem Biotechnol 1990. Grobbelaar JU.12:355–68. Carboxymethyl cellulose protects algal cells against hydrodynamic stress. J Biosci Bioeng 2001. Wiley. Ratledge C. 3.29:152–61. Chisti Y. Kulkarni MG. Humphreys K. Janssen M. J Am Oil Chem Soc 2006. Waste cooking oil — an economical source for biodiesel: A review. Metting FB. Microalgae. Metting B. Lee JW. In: Richmond A. Trends Biotechnol 2000. Biochem Eng J 2005.15: 377–90. 1753–69. Chisti Y. Adv Appl Microbiol 2002. The biochemistry and molecular biology of lipid accumulation in oleaginous microorganisms. J Appl Phycol 1996. Acién Fernández FG. Acién Fernández FG. editor. Fukuda H. Crit Rev Food Sci Nutr 1991. Photobioreactors: production systems for phototrophic microorganisms. J Ind Microbiol 1996.22:45–52. Vidya Sagar D. Prog Lipid Res 2006. García Camacho F. Biotechnology 1988. Seibert M. Appl Microbiol Biotechnol 2001. González Pascual V. Gallardo Rodríguez J. Knothe G. Production of methyl-ester fuel from microalgae. Llabres P. Chisti Y. Myers J. Lorenz RT. Harwood JL. 97–115. Naik SN. Xiong VF. Flynn T. Correia MJN.70:231–47. Molina Grima E. Microalgae as food and supplement. Biodiesel: the use of vegetable oils and their derivatives as alternative diesel fuels. and scaleup. Molina Grima E.27:1217–28. Plant Physiol 1953.6:331–5. Botryococcus braunii: a rich source for hydrocarbons and related ether lipids. J Environ Radioact 2005. Moo-Young M. Kapdan IK. Biometals 2002. growth and photosynthesis of Phaeodactylum tricornutum. Enzyme Microb Technol 2006. Enclosed outdoor photobioreactors: light regime. Cerón García MC. sulfate-limited chemostat system. McGraw-Hill. p. Philliphs JN. Blackwell. mass transfer. Neori A. Bagby MO. Growth rate of Chlorella in flashing light. Green alga hydrogen production: progress. Sánchez Mirón A. Belarbi E-H. Bordado JM. editors. Kosourov S. González-Ballester D. Dalai AK. Chisti Y. Grobbelaar JU. Camacho Rubio F.10:248–68. vol.26(5):487–94. Acién Fernández FG. Greenbaum E. P and metal removal: a review. Handbook of microalgal culture: biotechnology and applied phycology. Pulz O.18:506–11. Influence of high frequency light/dark fluctuations on photosynthetic characteristics of microalgae photo acclimated to different light intensities and implications for mass algal cultivation.74:3–16. Seibert M. Robles Medina A. Scale-up of tubular photobioreactors.83:823–33. Wynn JP. Mazzuca Sobczuk T. García Camacho F. mass culture methods. Photobioreactors: light regime. Trends Biotechnol 2004.38:569–82.4: 22–30. Molina Grima E. p.81:193–210. Biotechnol Adv 2005. Drew SW. et al. Biological principles of mass cultivation. In: Flickinger MC. Biotechnol Bioeng 2003. Single cell oils — have they a biotechnological future? Trends Biotechnol 1993. Considerations for designing bioreactors for shear-sensitive culture. Kargi F. Sánchez Mirón A.24–5:355–61. Bioprocess Biosyst Eng 2006. Appl Biochem Biotechnol 2005. Trends Biotechnol 2003. Raposo I. Technical aspects of biodiesel production by transesterification — a review. Mata-Alvarez J.17:477–89.8:325–33.6:1291–6. 1999. et al. J Phycol 1968. photosynthetic efficiency. Jelen's cost and optimization engineering. Chisti / Biotechnology Advances 25 (2007) 294–306 Fedorov AS. biocatalysis and bioseparation. Turbulence in algal mass cultures and the role of light/dark fluctuations. Transgenic microalgae as green cell-factories. Ratledge C.92:405–16. Nedbal L. Algal nutrition. Myers J. Acién Fernández FG. Meher LC. Ind Eng Chem Res 2006. Lemke P. Min DB. Gavrilescu M. Enzyme Microb Technol 2001.57:287–93. Chisti Y. Comp Rev Food Sci Saf 2005. and future prospects. Jang ES. 3rd ed. Biogas plants in Denmark: successes and setbacks.666:172–208. scale-up. Chisti Y. Biologically-active compounds from microalgae. García Camacho F. Pyne JW. Guschina IA.8:386–94. Shear effects on suspended marine sponge cells. Fernández E. Biotechnol Adv 2007.

editors. Bot Rev 2001. Terry KL. Yokoyama S-Y.50:431–65. characterization. Purton S. García Camacho F. et al. Energy Convers Manag 1995.43:33–40. Hader DP. Encyclopedia of bioprocess technology: fermentation. Chisti Y. Chisti Y. Biochem Eng J 2003. Contreras Gómez A.14:401–35. Bioreactors. Dunahay TG. J Biosci Bioeng 2006. Microalgal metabolites. Walter TL. Shear rate in stirred tank and bubble column bioreactors. Shimizu Y. Crit Rev Biotechnol 2005. Duran E. Chem Eng J 2006. Lee CI. Sánchez Mirón A.86:1097–107. p. Microalgal metabolites: a new perspective. Plant Cell Rep 2005. 1999. Vaishampayan A. Brown LM. Shimizu Y. Department of Energy's Aquatic Species Program — biodiesel from algae. Bubble column and airlift photobioreactors for algal culture. Benemann J. Genetic-engineering approaches for enhanced production of biodiesel fuel from microalgae. Biotechnol Bioeng 1986. Contreras Gómez A.124:1–5. Crit Rev Biotechnol 2004. Roessler PG. . Chisti / Biotechnology Advances 25 (2007) 294–306 Singh S.5:113–23. Shear stress tolerance and biochemical characterization of Phaeodactylum tricornutum in quasi steady-state continuous culture in outdoor photobioreactors. Chisti Y. Chem Eng Res Des 2003. Dunahay T. Joannis-Cassan C.16:287–97. Production. Chisti Y. Suresh B.46:1872–87. Sharma R. Comparative evaluation of compact photobioreactors for large-scale monoculture of microalgae.566:255–70.69:451–5.70:249–70. Sánchez Pérez JA. Microalgae as bioreactor. Gupta AK. Schneider JC. Vial C. biocatalysis and bioseparationWiley. In: Flickinger MC. Pharmaceuticals from cultured algae. Dey T. Thakur RK. Sheehan J. J Ind Microbiol 1990. Cerón García M-C. Collet C. Fernández Sevilla JM. 395–419. Lee SB. Curr Opin Microbiol 2003. A look back at the U. Moo-Young M. Jarvis EE. Schwartz RE. Dote Y. J Biotechnol 1999. photo.25:73–95. Annu Rev Microbiol 1996. Molina Grima E.S. Sánchez Mirón A.24:97–124. Biotechnol Adv 2001. García Camacho F.81:787–826.36: 729–31. and applications of lipases. Ravishankar GA. Report NREL/TP-580–24190. System design for the autotrophic production of microalgae. 1998. Molina Grima E. Heacox DA. Nauman EB. Bioactive compounds from cyanobacteria and microalgae: an overview.28:988–95. Carbon dioxide fixation by algal cultivation using wastewater nutrients. CO.6: 236–43. Cyanobacterial biofertilizers in rice agriculture. Becker DK. Golden. Djelveh G. Sawayama S. Chisti Y. Drew SW. ACS Symp Ser 1994. Spolaore P. Moo-Young M. Biodiesel processing and production. Commercial applications of microalgae. Sinha RP. Terry KL. Static mixers in the process industries — a review. Zhang Z. Phytoremediation — a novel and promising approach for environmental clean-up.306 Y. Enzyme Microb Technol 1985. Bhan U. Contreras Gómez A. Zhang Z. Tredici MR.67: 453–516. Isambert A. Van Gerpen J. Photosynthesis in modulated light: quantitative dependence of photosynthesis enhancement on flashing rate. CO2 fixation and oil production through microalga. Yun YS. Casas López JL. Biotechnol Adv 1996. Roessler P.19: 627–62. Banerjee UC. Banerjee UC.101:87–96. et al.7:474–87. Kate BN. Plasmid stability in recombinant Saccharomyces cerevisiae. Sánchez Mirón A. Park JM. Yang JW. Effects of the hydrodynamic environment and shear protectants on survival of erythrocytes in suspension. Molina Grima E. AIChE J 2000. Nigam KDP. Raymond LP. Fuel Process Technol 2005. García Camacho F. J Biotechnol 1995.24:629–41. purification. Inoue S. National Renewable Energy Laboratory. Chisti Y. J Chem Technol Biotechnol 1997. Rodríguez Porcel EM. Chisti Y.