Rev Environ Sci Biotechnol (2011) 10:3141 DOI 10.

1007/s11157-010-9214-7

MINI REVIEW

Commercial application of microalgae other than as biofuels: a brief review

John J. Milledge

Published online: 19 August 2010 Springer Science+Business Media B.V. 2010

Abstract There has been considerable discussion in recent years about the potential of micro-algae for the production of sustainable and renewable biofuels, but there may be other more readily exploitable commercial opportunities for microalgae. This paper briey reviews the current and potential situation for the commercial application of the growth of microalgae for products other than biofuels. Keywords Algae Microalgae Commercial Application Biofuel Algal Food Carotenoid Phycobiliprotein Biochemical Market

1 Introduction There has been considerable recent interest in the production of biofuels from micro-algae. At present the process of producing fuel from algae would appear to be uneconomic with over 50 algal biofuel companies and none as yet producing commercial-scale quantities at competitive prices (Milledge 2010; Pienkos and Darzins 2009). Currently algal biofuel

J. J. Milledge (&) Visiting Research Fellow, School of Civil Engineering and the Environment, University of Southampton, Lanchester Building (7), Higheld, Southampton SO17 1BJ, UK e-mail: j.milledge@soton.ac.uk

is not competitive to other biofuels and cost of production needs to be reduced signicantly to become economic (Clarens et al. 2010; Wijffels 2007). The European Algae Biomass Association has estimated that it may take another 10 to 15 years to turn laboratory experiments into industrial-scale production of algal biofuel (Kovalyova 2009). Although the commercial exploitation of microalgal biofuel may be some time off It is generally agreed that microalgae have a great potential to produce a wide range of important biochemicals for food, medical, research and other uses and that many exciting and important biochemicals are yet to be discovered from microalgae (Becker 1994; Radmer and Parker 1994; Gavrilescu and Chisti 2005; Singh et al. 2005; Spolaore et al. 2006; Wijffels 2007; Raja et al. 2008). The use of microalgae by humans dates back thousands of years to the Chinese who used Nostoc to survive famine. Other species of micro-algae including, the blue green algae species, Arthrospira (Spirulina) and Aphanizomenon have also been used by humans for thousands of years (Singh et al. 2005; Spolaore et al. 2006). Spirulina has been exploited by ancient peoples in both Chad and Mexico as a source of food (Chisti 2006; Henrikson 2008). Although, microalgae have been used for food by humans for thousands of years, microalgae culture is one of the modern biotechnologies. Uni-algal cultures were rst achieved in 1890 with Chlorella vulgaris and the use of this type of cultures was used for

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the study of plant physiology in the early 1900 (Borowitzka 2006). In 2004 the global market for microalgal biomass was estimated to be 5,000 t of dry matter per year and generated a turnover of US$ 1,250 million (Spolaore et al. 2006). Microalgae contain about 50% carbon in their biomass and this carbon is obtained in most cases from atmospheric carbon dioxide and therefore microalgae are attracting interest as vehicles for carbon sequestration for industrial processes (Nakamura et al. 2001; Benemann et al. 2003; Huesemann et al. 2003; Ono and Cuello 2003; Weissman and Benemann 2003; Chisti 2006). (Fig. 1). The use of algae for therapeutic purposes has a long history, but the search for biologically active substances from algae, especially examination for antibiotic activity began only in the 1950s. Much of that laboratory work up until the 1980s focused on macroalgae (Borowitzka 1995). Approximately, 15,000 natural marine products have now been screened for biological activity and 45 marine derived natural products have been tested to be used as medical drugs in preclinical and clinical trials. Only 2 have been developed into registered drugs, one from a marine snail and the other from a sea squirt, although none as

yet from microalgae (Wijffels 2007). It is reported that preclinical trials are currently being undertaken on an anti-cancer drug, Curacin, derived from the blue-green algae, Lyngbya majuscula (Day 2009). There have also been reports of the use of products derived from algae being used to combat HIV infection, but it would appear that it is 5 years from commercialisation (Anitei 2007; BBC 2007). Microalgae are responsible for over 50% of primary photosynthetic productivity on earth and are budding sunlight factories for a wide range of potentially useful products, but are barely used commercially (Chisti 2000; Gavrilescu and Chisti 2005; Chisti 2006; Wijffels 2007). In the early 1950s the increase in world population lead to the search for new alternative food and protein sources and algae appeared at the time a good candidate (Huntley and Redalje 2007; Spolaore et al. 2006). Although, algae have and are still used for food, to some extent around the world, the large scale production of algae to solve the worlds food calorie and protein shortage has not materialised. The large-scale cultivation of microalgae and the use of its biomass for the production of useful products were rst considered seriously in Germany during World War II (Becker 1994).

Fig. 1 Trial Algal race-way growth ponds using Carbon Dioxide from a power plant. Courtesy Seambiotic, Israel

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Commercial largescale modern algae culture started in the early 1960s in Japan with the culture of Chlorella. In the early 1970s culturing and harvesting of Spirulina began in Mexico. The third major area of commercialisation of algae occurred, in Australia, with the growth of Dunaliella salina for the production of b-carotene. Plants were then subsequently built in the USA and Israel and production of blue-green algae also commenced in India. Plants have recently been built in the USA and India for the growth of Haematococcus pluvialis as a source of astaxanthin, approved as food colouring and also a powerful anti-oxidant (Borowitzka 2006; Spolaore et al. 2006). A common feature of most of the algal species currently produced commercially (i.e. Chlorella, Spirulina and Dunaliella) is that they grow in highly selective environments which means that they can be grown in open air cultures and still remain relatively free of contamination by other algae and protozoa. Thus, Chlorella grows well in nutrient-rich media, Spirulina requires a high pH and bicarbonate concentration and Dunaliella salina grows at very high salinity. Those species of algae which do not have environmental selective advantages may need to be grown in closed systems. This includes most of the marine algae grown as aquaculture feeds (e.g. Skeletonema, Chaetoceros, Thalassiosira, Tetraselmis and Isochrysis) and the dinoagellate, Crypthecodinium cohnii, grown as a source of long-chain polyunsaturated fatty acids, as well as almost all other species being considered for commercial mass culture (Borowitzka 2005; Borowitzka 2006; Chisti 2006; Huntley and Redalje 2007).

2 Current commercial uses of algae 2.1 Health foods Chlorella is produced by more than 70 companies with the largest producer, Taiwan Chlorella Manufacturing and Co producing 400 t of dry algal biomass per year (Spolaore et al. 2006). Chlorella is sold as a health food or dietary supplement (Borowitzka 2006). Several reports from Japan have described various potential therapeutic effects of Chlorella and although these are encouraging ndings, the investigations were initiated by the

Chlorella producing companies and must be viewed as such (Becker 1994) Suggested health benets including efcacy on gastric ulcers, wounds and constipation together with preventive action against both atherosclerosis and hyper-cholesterol and antitumor activity. The suggested most important active substance is b-1,3-glucan which is believed to be an active immune-stimulator, free radical scavenger and a reducer of blood lipids. Unfortunately, the American Cancer Society concluded, however, available scientic studies do not support its effectiveness for preventing or treating cancer or any other disease in humans (American Cancer Society, Inc. 2007). Spirulina (Arthrospira) is used in human nutrition because of it high protein content and excellent nutrient value (Spolaore et al. 2006). It is also a valuable source of the essential fatty acid, linolenic acid that cannot be synthesised by humans (Becker 1994). A wide range of medical benets for a broad range of conditions have been claimed, but the full validations of many of these claims is still awaited (Becker 1994; U.S. Department of Health and Human Services 2001; Singh et al. 2005; Spolaore et al. 2006; American Cancer Society, Inc. 2007; Henrikson 2008). Many companies are producing nutraceuticals (food supplements with claimed nutritional and medicinal benets) made from Spirulina. DIC claim to be the biggest manufacture of Spirulina in China (DIC 2009). Production of Spirulina in Hainan by DIC was estimated at 300 t per annum (Lee 1997). The largest plant is Earthrise Farms in California, USA covering over 444,000 m2 producing algal tablets and powder sold in over 20 countries and is owned by DIC in Japan (Earthrise Nutritionals LLC 2004). Cyanotech, in Kona, Hawaii, produces a powder under the name Spirulina Pacica (Cyanotech Corporation 2007). The market for dried Spirulina was estimated to be US$ 40 million in 2005 (Singh et al. 2005). (Fig. 2). 2.2 Carotenoids Algae contain carotenoids, yellow orange or red pigments, that include b-carotene a substance converted by the body to Vitamin A. There are over 400 known carotenoids, but only a few are used commercially, the two main compounds being b-carotene and astaxanthin. The most important uses of carotenoids

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Fig. 2 Arial view of Cyanotech Algal Production Facility showing race-way growth ponds. Courtesy Cyanotech Corporation, Hawaii, USA

are as food colourants and as supplements for human and animal feeds (Becker 1994; Spolaore et al. 2006). The average concentration of carotenoids in most algae is only 0.12%, but Dunaliella when grown under the right conditions of high salinity and light intensity will produce up to 14% b-carotene (Becker 1994; Singh et al. 2005; Borowitzka 2006; Spolaore et al. 2006; Wijffels 2007). Dunaliella is, therefore, well suited to the commercial production of b-carotene and several industrial production plants are in operation around the world including Australia, Israel, USA and China (Spolaore et al. 2006). The major producer is Cognis Nutrition and Health, whose farms cover 800 ha in Western Australia and produce b-carotene extracts together with algal powder for human and animal use. The prices of these products in 2004 varied from US$300 to US$3000/kg (Spolaore et al. 2006; Cognis 2008). (Fig. 3). Until the 1980 production of b-carotene was synthetic. Natural carotenoids, although more expensive than synthetic have the advantage of supplying the natural isomers in their natural ratio and the natural isomers of b-carotene are considered superior to the trans synthetic form (Spolaore et al. 2006; Singh et al. 2005). In 1994 algal b-carotene was sold

in small quantities due to the cost and the majority of production was synthetic, however it was concluded that the increasing number of algal plants producing b-carotene rich Dunaliella, at that time, may change the situation (Becker, 1994). It has since been reported that b-carotene from Dunaliella is a substantial growing industry and commercial utilisation is economically viable (Singh et al. 2005: Chisti 2006). Astaxanthin is another carotenoid that can be derived from algae and is principally used in sh farming and as a dietary supplement or anti-oxidant. The annual worldwide aquaculture market for this pigment in 2004 was estimated to be US$ 200 million with an average price of US$ 2500/kg, but the market is dominated by the synthetic form. Astaxanthin can be produced by Haematococcus, a freshwater alga that normally grows in puddles, birdbaths and other shallow fresh water depressions. Haematococcus can contain up to 3% astaxanthin, but it requires a two stage culture process which is not suited to open pond cultivation. The rst stage of the process is designed to optimise algal biomass (green-thin walled agellated stage with optimum growth at a temperature 2225C) and the second stage (thick walled resting stage) under intense light and nutrient poor conditions during which astaxanthin is produced (Borowitzka 2006;

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Rev Environ Sci Biotechnol (2011) 10:3141 Fig. 3 Cognis Algal Growth Lagoons, Whyalla, Australia. Courtesy of Cognis Australia Pty Ltd

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Spolaore et al. 2006). Due to its high price, the astaxanthin produce from Haematococcus cannot compete with synthetic forms (Spolaore et al. 2006; Wijffels 2007). However, for some applications natural astaxanthin is preferred for example in carp, chicken and red sea bream diets due to enhanced natural pigment deposition, regulatory requirements and consumer demand for natural products (Spolaore et al. 2006). Commercial production is being carried out in Hawaii, India and Israel, where Algatech sell a crushed Haematococcus biomass on the pharmaceutical market (Algatech 2004; Borowitzka 2006; Spolaore et al. 2006). Cyanotech, in Hawaii claimed a market share of over 95% of the animal nutrition market for algae-based astaxanthin products, but subsequently stopped selling into the animal nutrition market in March 2008. (Cyanotech Corporation 2008). (Fig. 4) 2.3 Phycobiliproteins In addition to chlorophyll and the lipophilic pigments, such as the carotenoids, certain types of microalgae especially red algae, or rhodophyta, contain phycocyanin and Phycoerythrin (Becker 1994). These photosynthetic accessory pigments, collectively known Phycobiliproteins, are deeply coloured (red or blue), water soluble, complex, proteinaceous

compounds. These algae pigments have the potential as natural colourants for food, cosmetics and pharmaceuticals. Dainippon Ink and Chemicals produces a blue food colourant from Spirulina, called Lina blue, that is used in chewing gum, ice slush, sweets, soft drinks, dairy products and wasabi (Spolaore et al. 2006; DIC Corporation, 2008; Raja et al. 2008). Phycobiliproteins can be commercially produced from Spirulina and the red microalgae Porphyridium and Rhodella (Becker 1994; Singh et al. 2005; Borowitzka 2006; Spolaore et al. 2006). Phycobiliproteins are widely used in clinical or research immunology because they are very powerful and highly sensitive uorescent properties (ProZyme 1999; Spolaore et al. 2006; Raja et al. 2008). In 1997 the global market for Phycobiliproteins colourants was estimated at US$ 50 million and prices vary from US$ 3 to US$ 25/mg (Spolaore et al. 2006). 2.4 Fatty acids Humans and animals lack the requisite enzymes to synthesise polyunsaturated fatty acids (PUFAs) of more than 18 carbon atoms and they must obtain them from food and are, therefore, often known as essential fatty acids. A group of essential fatty acids which are attracting a lot of attention currently are known as omega-3, a group of unsaturated fatty acids

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Fig. 4 Haematococcus being grown in a tubular photobioreactor. Courtesy Seambiotic Israel

where a carbon double bond is in the third position from the methyl or omega end (British Nutrition Foundation 2000; Spolaore et al. 2006; Wijffels 2007). Oily Fish and sh oils are well known sources of PUFAs, but issues have been raised concerning possible accumulation of toxins in sh (Food Standards Agency 2008). Fish obtain PUFAs from algae, with marine algae, such as diatoms, being a particularly rich source and therefore algae have been proposed as a commercial source (Borowitzka 2006; Spolaore et al. 2006). Docosahexaenoic acid (DHA) produced by heterotrophic (using plant or animal materials as an energy source rather than light in photosynthesis (autotrophic)) culture of the dinoagellate, Crypthecodinium cohnii, is the only currently commercial produced fatty acid from algae (Borowitzka 2006; Spolaore et al. 2006). DHA (22: 6) is a 22 carbon chain with six cis double bonds; the rst double bond is located at the third carbon from the omega end (an omega-3 fatty acid). It is used as a supplement in infant formulas and as a dietary supplement. It is essential for the proper functioning of our brains as adults, and for the development of our nervous system and visual abilities during the rst 6 months of life. In addition,

omega-3 fatty acids are part of a healthy diet that helps lower risk of heart disease (University of Maryland Medical Centre 2007). Although, infants that are breastfed should receive enough DHA, if the mother has an adequate intake of this fatty acid, many organisations have suggested that DHA should be added to baby milk formula (Koletzko et al. 2005; Spolaore et al. 2006; University of Maryland Medical Centre, 2007). The world wholesale market for infant formula in 2005 was estimated to be about US$ 10 billion per annum (Spolaore et al. 2006). Martek produce DHA using Crypthecodinium cohnii for baby formula and in 2003 production was 240 tons and it is now a company with more than 525 employees and revenue of more than US$ 300 million. Martek acquired, OmegaTech, another producer of a DHA, oil known as DHA gold, an adult dietary supplement from Schizochytrium (Spolaore et al. 2006; Martek Biosciences 2008). Nutrinova, formerly known as Hoechst until 1997 when it was taken over by Celanese, produced an oil containing DHA from Ulkenia. In 2005 Lonza, based in Switzerland, acquired Nutrinovas DHA business and sells the DHA oil as a vegetarian source of omega-3 (Food

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Standards Australia New Zealand 2004; Lonza 2005; Spolaore et al. 2006; Anon 2008). A process for producing high-purity Eicosapentaenoic acid, EPA, another omega-3 fatty acid (20:5), from Phaeodactylum tricornutum has been developed by the University of Almeria in Spain. An economic analysis, on a potential facility producing 430 kg 96% pure EPA per year, estimated the cost total cost of production at US$ 4,602/kg, with 60% of the cost arising from the recovery process and 40% from the biomass production. It is believed that the cost needs to be reduced by 80% to be economically viable. The residual biomass following the extraction of the EPA contains too much residual solvent to be sold for animal feed and therefore must be incinerated (Molina-Grima et al. 2003). The annual worldwide demand of EPA is 300 t (Singh et al. 2005). The production of EPA from Nannochloropsis and, the diatom, Nitzschia is reportedly under study (Borowitzka 2006; Spolaore et al. 2006). It is reported that studies are being undertaken to produce omega-6 PUFAs; Linolenic acid (18:3) from Arthrospira and Arachidonic acid (20:4) from Porphyridium (Borowitzka 2006; Spolaore et al. 2006). 2.5 Stable isotopic biochemicals Microalgae are well suited as a source of isotopically labelled compounds due to their ability to incorporate stable isotopes from relatively inexpensive inorganic molecules into high value isotopic organic chemicals (Raja et al. 2008). The market for these chemicals is in excess of US$ 13 million (Spolaore et al. 2006). Spectra Stable Isotopes, now part of Cambridge Isotope Laboratories, sells marked amino acids at up to US$ 5900/g and marked nucleic acids at US$ 28/ mg (Spolaore et al. 2006; Cambridge Isotope Laboratories 2008). 2.6 Animal feed

(Spolaore et al. 2006). Approximately a dozen types are produced in relatively small quantities for the aquaculture industry (Chisti 2006). Many studies have shown the suitability of algae as a potential animal feed and as a replacement for conventional protein sources such as soybean and sh meal (Becker 1994; Spolaore et al. 2006). Algae are the normal natural food for many animals used in aquaculture and it is not surprising that they are considered the best food source for aquaculture. Unfortunately, the trend is to avoid using live algae due to their high cost and production difculties. The cost of producing dry algal biomass feed, in Australia, varies from US$ 80/kg to US$ 800/kg (Becker 1994; Spolaore et al. 2006). Other cost estimates have given costs between US$ 50/kg to US$ 150/kg with a peak value of US$ 1000/kg (Raja et al. 2008). Yeast can be used as a replacement feed, but the omission of algae from aquaculture may give less predictable performance and the total replacement of algae in aquaculture diets are not yet considered sufciently advanced for widespread utilisation (Becker 1994; Spolaore et al. 2006). In addition to direct feed, microalgae can be used as a feed source for zooplankton which can in turn be used as a feed for sh (Raja et al. 2008). One problem that has been in encountered is, with the notable exception of Spirulina, poor digestibility due to the high content of cellulose cell wall material. Ruminates, such as sheep and cattle, are capable of digesting cellulose material and it is therefore possible to feed algae direct to them, but this has not gained much commercial favour yet (Becker 1994). Poultry can be fed up to 510% algae and this can have a positive effect on the development of colour within the skin and egg yolk due to the carotenoids (Becker 1994; Borowitzka 2006). Higher concentrations of algae in the feed can lead to adverse effects (Spolaore et al. 2006). 2.7 Human food

Microalgae are an important food source and feed additive in the commercial rearing of many aquatic animals (Borowitzka 2006). Over 30% of the current world algal production is sold for animal feed and over 50% of the world production of Spirulina is used as feed supplements. In 1999 the production of microalgae for use in aquaculture reached 1000t

Although, microalgae are eaten as a food in China and Chad and had been considered as a solution to the worlds food shortage, their use on a global scale appears limited to health food and food supplements (Becker 1994; Borowitzka 2006; Spolaore et al. 2006). (Table 1)

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38 Table 1 The current state of microalgal production (Spolaore et al. 2006) Alga Spirulina Chlorella Dunaliella Aphanizomenon Haematococcus Crypthecodinium Schizochytrium Annual production 3,000 t dry wt 2,000 t dry wt 1,200 t dry wt 500 t dry wt 300 t dry wt 240 t DHA oil 10 t DHA oil Producer country China, India, USA, Myanmar, Japan Taiwan, Germany, Japan Australia, Israel, USA, China USA USA, India, Israel USA USA

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Applications and products Human & animal nutrition, phycobiliproteins, cosmetics Human nutrition, aquaculture, cosmetics Human nutrition, cosmetics, b-carotene Human nutrition Aquaculture, Astaxanthin DHA oil DHA oil

3 Future development of micro-algal applications 3.1 Market potential Microalgae have been exploited by man for millennia and the BEAM network, supported by Murdoch University, Australia, believes that microalgae biotechnology has grown and diversied signicantly over the past 30 years (Borowitzka 2006). Although, some progress has been made and there are commercial microalgae applications, including pigments, fatty acids and health foods, only a few hundred of the thousands of species of microalgae have been studied and just a handful are cultivated on an industrial scale (Spolaore et al. 2006). It would appear that the potential of commercial applications of microalgae are enormous, but that despite development over last 50 years the number of commercially available products are still fairly limited and although there are a number of types of closed bioreactor being investigated and available, the majority of microalgal production is in open ponds. The main commercial product, despite the enormous range of biochemicals potentially available from microalgae, appears to be health food that may produce health benets, but may be subject to fashion and fad. The second current key area of commercial application of microalgae appears to be food additives in form of carotenes, pigments and fatty acids. These can have functional advantages over synthetic products or products from other natural sources, but can be at a cost disadvantage. The challenge in the application of microalgae for commercial ends is to focus only on those products with a large market and or prot potential where the use of microalgae leads to clear competitive

advantages (Radmer and Parker 1994). Fuel and food would appear to offer the largest markets. The growth of microalgae to solve the world food shortage has in the past been considered, but the wide scale commercial growth of algae for human food is restricted mainly to health foods, food supplements and food additives. The health food market is the branch of algae production with the highest sales, but the market is dependent on a number of claims of health benets without the necessary scientic proof of efcacy (Becker 1994; American Cancer Society, Inc 2007). There has also been some market sentiment that Spirulina was being over produced (Lee 1997). Food additives, such as b-carotene, pigments and fatty acid, from microalgae can be superior to synthetic products and products from other natural sources, such as sh oil, but unfortunately they are often also considerably more expensive. There appears to be very considerable potential for the discovery of new therapeutic biochemicals from microalgae, but an immediate breakthrough for algae based products does not appear imminent (Becker 1994; Singh et al. 2005; Spolaore et al. 2006). Current commercial viable exploitation of microalgae products is currently limited to products other than fuel and the immediate future for the commercialization of microalgae may be with non fuel products, but the lessons learned from microalgal non fuel products together with their potential coproduction with fuel may lead to the more rapid commercial realisation of microalgal biofuel. 3.2 Growth systems The commercial growth of algae in open ponds, despite over 30 years of research, is still only currently viable

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for three taxi, Spirulina, Dunaliella and Chlorella, mainly due to the suppression of the growth of competitive species by use of highly selective environments (Huntley and Redalje 2007). Currently the majority of microalgal production occurs in outdoor ponds (Spolaore et al. 2006). Concerns have also been expressed about the possible contamination of food from microalgae grown in open ponds (Becker 1994). Bioreactors are considerably more expensive than open ponds and their use will probably be restricted to very high value products (Becker 1994). It has been argued that lower extraction costs, due to the higher algae content, can make bioreactors more competitive (Chisti 2007). A study on the production of the valuable fatty acid, EPA, found that 60% of the costs arise from the recovery process and the cost of algae biomass production in bioreactors is high and need to be reduced (Molina-Grima et al. 2003). It has also been concluded that, although some products are now being produced in bioreactors commercially, the development of microalgae biotechnology has been slowed by the limited performance of bioreactors (Spolaore et al. 2006). Open ponds are likely to remain the major means of production, but efcient closed bioreactors may be viable in the production for high value products where purity is essential or a sensitive algae species is required. 3.3 Species selection Many consider that the genetic modication of microalgae is the best way to improve the yield of valuable products at reduced cost (Becker 1994; Sheehan et al. 1998; Spolaore et al. 2006; Chisti 2007). The absence of cell differentiation in microalgae can make genetic manipulation simpler than in higher plants, but progress on the genetic engineering of microalgae was relatively slow until recently (Raja et al. 2008). The NREL in the USA spent considerable time, effort and resources on the genetic modication of microalgae (Sheehan et al. 1998). The NREL work on genetic modication may have diverted their resources from algae selection and process optimisation that may have yielded more commercial benet. Genetic modication has a considerable image problem, particularly in Europe and genetically modied algae may also be seen as a potential environmental threat, particularly if open pond systems are to be used. Part of the appeal of microalgae substances for

food and therapeutic use is their natural image and genetic modication may have a negative effect on this. Algae strain selection and process optimisation rather than genetic engineering may be the key areas for future development (Wijffels 2007).

References
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