C H A P T E R
1
Marine Microalgae Biotechnology:
Present Trends and Future Advances
Jayachandran Venkatesan, Panchanathan Manivasagan,
Se-Kwon Kim
Pukyong National University, Department of Marine-Bio Convergence Science and Marine Bioprocess
Research Center, Busan, South Korea
1. INTRODUCTION
2. ISOLATION AND CULTURE
Microalgae are unicellular species, commonly
found in marine and freshwater with the size
ranging from a few micrometers to a few hundreds of micrometers. It has been estimated
that 2 105 to 8 105 species exist. Microalgae
are a promising source for several bioactive
compounds (Cardozo et al., 2007; Norton et al.,
1996). Polymers, peptides, fatty acids, carotenoids, toxins, and sterols are important bioactive
products produced by microalgae. Microalgae
do not contain stems and roots as do higher
plants. The three most important classes of
microalgae in terms of abundance are the diatoms (Bacillariophyceae), the green algae (Chlorophyceae), and the golden algae (Chrysophyceae).
The cyanobacteria or blue-green algae (Cyanophyceae) are also referred to as microalgae, i.e.,
Spirulina (Arthrospira platensis and Arthrospira
maxima).
Isolation, production, and culture of microalgae are important steps to producing a commercial product in sufficient amounts. Microalgae
growths are dependent on light, water temperature, nutrient concentration, salinity, and pH
(Brennan and Owende, 2010; Mutanda et al.,
2011). Collection, sampling, and preservation
techniques must be optimized to grow a highquality microalgae sample. Three important
isolation techniques are commonly used for
microalgae collection:
Handbook of Marine Microalgae
http://dx.doi.org/10.1016/B978-0-12-800776-1.00001-7
1. Streaking
2. Serial dilution
3. Single-cell isolation (He et al., 2012).
Different kinds of culture techniques are
available to culture microalgae; culture media
and culture conditions are the main aspects to
be considered in microalgae cultivation.
1
© 2015 Elsevier Inc. All rights reserved.
2
1. MARINE MICROALGAE BIOTECHNOLOGY: PRESENT TRENDS AND FUTURE ADVANCES
Several important nutrients (nitrate, urea,
ammonium, vitamins, phosphorous, nitrogen,
iron, manganese, selenium, cobalt, nickel, and
zinc) are required for the production of any
microalgae species (Andersen, 2005; Harrison
and Berges, 2005).
Microalgae can be cultured mainly in two
different kinds of systems:
1. Open cultivation
2. Photobioreactor.
The traditional open cultivation method
(lakes and ponds) has been used since 1950.
The photobioreactor system has more advantages than open cultivation, avoiding several
issues such as contamination and environmental
disturbance. There are three distinct algae production mechanisms: photoautotrophic, heterotrophic, and mixotrophic.
Phaeodactylum, Chaetoceros, Nannochloropsis,
Skeletonema, and Thalassiosira genera (Brown
et al., 1996; Spolaore et al., 2006). Marine algae
have been used as food additives, for bivalve
mollusks (oysters, scallops, clams, and mussels)
and fishmeal. There is a promising work in the
area of using microalgae as a food additive to
increase weight, oil content, and protein deposition in muscle. The nutritional composition of
microalgae is made up of carbohydrates, proteins, vitamins, lipids, antioxidants, and other
trace elements. Kang et al. (2012) reported the
nutritional composition of microalgae (Navicular
incerta): crude lipidse8.76%, crude proteinse
50.38%, and carbohydratese10.84%, respectively. A general overview of microalgae in
food usage is explained in Figure 1.
3.2 Fatty Acids
3. APPLICATIONS OF MARINE
MICROALGAE
Microalgae have been widely used for various
applications including human and animal nutrition, cosmetics, pharmaceuticals, CO2 capture,
bioenergy production, and nutrient removal
from wastewater. Biological properties of algae
and their components are well studied in the
following areas of research: antioxidants, antimicrobials, anticancer agents, anti-inflammatory
and cardiovascular health, anti-obesity, and antidiabetic activity (Dominguez, 2013).
3.1 Animal Feed
Microalgae have been investigated as human
and animal foods for over six decades (Liu
et al., 2014). Although several hundreds of
microalgae species have been investigated for
food applications, only a few have been
used in aquaculture. The most common species
are Chlorella, Tetraselmis, Isochrysis, Pavlova,
Microalgae are traditionally considered
good sources of fatty acids (Benemann, 1989;
Borowitzka, 2013). The accumulation of fatty
acids by microalgae is well developed and
presented elsewhere (Griffiths and Harrison,
2009; Rodolfi et al., 2009). The presence of
eicosapentaenoic acid (C20:5) and docosahexaenoic acid (C22:6) is of interest for their health
benefits (Harris et al., 2008; Mozaffarian and
Rimm, 2006). Cyanobacterium Spirulina is rich
in linolenic acid and a good source for polyunsaturated fatty acid (PUFA) (Mahajan and
Kamat, 1995).
3.3 Nutraceuticals
Microalgae are particularly of interest as a
source of nutraceuticals because algae can
produce a number of biomolecules, viz, betacarotene, lutein, astaxanthin, chlorophyll,
phycobiliprotein, and PUFAs, which are useful
for human and animal health and development.
Marine microalgae pigments are carotenoids,
chlorophylls, and phycobiliproteins, which have
3. APPLICATIONS OF MARINE MICROALGAE
3
FIGURE 1 Food commodities from microalgae (Draaisma et al., 2013).
health-promoting properties such as vitamin precursors, antioxidants, immune enhancers, and
anti-inflammatory agents. Accordingly, microalgae pigments can find commercial applications
as innovative functional ingredients in the food
and feed industries, as well as in pharmaceuticals
and in cosmetics (Borowitzka, 2013). Microalgae
that have become more prevalent in food supplements and nutraceuticals are Nostoc, Botryococcus, Anabaena, Chlamydomonas, Scenedesmus,
Synechococcus, Perietochloris, and Porphyridium
because they contain vitamins and essential
elements such as potassium, zinc, iodine, selenium, iron, manganese, copper, phosphorus,
sodium, nitrogen, magnesium, cobalt, molybdenum, sulfur, and calcium. Algae are also high
producers of essential amino acids and omega 6
(arachidonic acid) and omega 3 (docosahexaenoic acid, eicosapentaenoic acid) fatty acids
(Simoons, 1990; West and Zubeck, 2012).
Average nutritional compositions of the
microalgae expressed as g per 100 g dry weight
are presented in Table 1.
3.4 Cosmeceuticals
Microalgae-derived skin care products have
been developed in the form of anti-aging
creams, refreshing or regenerating care
products, emollients and anti-irritants in peelers,
sunscreen creams, and hair care products. These
cosmetic products have been developed with
marine microalgae extracts or bioactive
components. By the early 2000s, numerous
cosmetic companies in Europe and the
United States started to launch cosmetics that
used extractions of microalgae such as Spirulina,
Chlorella, Arthrospira, Anacystis, Halymenia,
Nannochloro, and Dunaliella which act on the
epidermis to erase vascular imperfections by
boosting
collagen
synthesis
and
thus
possibly prevent wrinkle formation (Stolz and
4
1. MARINE MICROALGAE BIOTECHNOLOGY: PRESENT TRENDS AND FUTURE ADVANCES
TABLE 1 Average Nutritional Composition of the Microalgae Expressed as Grams per 100 g Dry Weight (West and
Zubeck, 2012)
Component
Spirulina
Dunaliella
Haematococcus
Chlorella
Aphanizomenon
Protein
63
7.4
23.6
64.5
1
Fat
4.3
7
13.8
10
3
Carbohydrates
17.8
29.7
38
15
23
Chlorophyll
1.15
2.2
0.4 (red), 1.1 (green)
5
1.8
Magnesium
0.319
4.59
1.14
0.264
0.2
Beta-carotene
0.12
1.6
0.054
0.086
0.42
Vitamin B1 (thiamin)
0.001
0.0009
0.00047
0.0023
0.004
Vitamin B2 (riboflavin)
0.0045
0.0009
0.0017
0.005
0.0006
Vitamin B3 (niacin)
0.0149
0.001
0.0066
0.025
0.013
Vitamin B5
(pantothenic acid)
0.0013
0.0005
0.0014
0.0019
0.0008
Vitamin B6 (pyridoxine)
0.00096
0.0004
0.00036
0.0025
0.0013
Vitamin B9 (folic acid)
0.000027
0.00004
0.00029
0.0006
0.0001
Vitamin B12
(cobalamine)
0.00016
0.000004
0.00012
0.000008
0.0006
Obermayer, 2005). Mycosporine-like amino
acids from Spirulina, Chlorella, and Dunaliella
are known to act as sunscreens to reduce ultraviolet (UV)-induced damage (Atkin et al., 2006;
Balskus and Walsh, 2010; Dionisio-Se Se, 2010;
Garciapichel et al., 1993; Priyadarshani and
Rath, 2012). Carotenoids such as astaxanthin,
lutein, zeaxanthin, and canthaxanthin found in
Haematococcus and Dunaliella protect against
sun damage and also are reported to have antioxidant activity (Gierhart and Fox, 2013; Guerin
et al., 2003; Tominaga et al., 2012; Walker et al.,
2005). Beta-carotene and astaxanthin are important microalgal products and are commonly
used in cosmetic applications (as protection
against oxidation of essential PUFAs, protection
against UV light effects) (Lorenz and Cysewski,
2000).
3.5 Pharmaceuticals
Marine microalgae are rich in biologically
active compounds, which can be used for pharmaceutical and nutraceutical development. Microalgae have a capacity to produce toxins that can be
used for pharmaceutical applications. Cyanobacteria are known to produce extracellular and
intracellular metabolites, which possess antifungal, antibacterial, and antiviral properties
(Baquero et al., 2008; Noaman et al., 2004;
Thillairajasekar et al., 2009). Figure 2 shows the
important compounds of blue green alga Lyngbya majuscula which can be used for drug
development.
1. Cytotoxic activity is important in anticancer
drugs (Sirenko and Kirpenko, 1999; Simmons
et al., 2005)
5
3. APPLICATIONS OF MARINE MICROALGAE
Me
S
H N
OMe
H 3C
O
H CH3 O
H
NH2
N
O
Me
H
Curacin A
O
H
OCH3
Majusculamide A
H 3C
N
H
N
O
OH
HO
O
N
H
HO
O
O
H 3C
O
O
O
OH
Lyngbyatoxin A
FIGURE 2
OCH3
Debromoaplysiatoxin
Variant structural types found in an identical species of the blue-green alga Lyngbya majuscula.
2. Antiviral activities are found mainly in
cyanobacteria but also in apochlorotic
diatoms and the conjugaphyte Spirogyra,
where certain sulfolipids are active, for
example, against the herpes simplex virus
(Muller-Feuga et al., 2003)
3. Antimicrobial activity is under investigation
to find new antibiotics, although currently
the success rate is about 1% (Muller-Feuga
et al., 2003)
4. Antifungal activity is found in different
extracts of cyanobacteria (Nagai et al., 1992)
5. Neuroprotective agents from Spirulina
(Chamorro et al., 2006; McCarty, 2007;
Nuhu, 2013)
6. Antioxidants (Miranda et al., 1998)
7. Anti-inflammatory activity (Jin et al., 2006)
8. Cardiovascular health effects (Doughman
et al., 2007)
9. Gastric and hepatic protective effects (AbdelWahhab, Ahmed & Hagazi, 2006)
10. Antidiabetic and antiobesity properties
(Mayer and Hamann, 2005)
11. Antiviral activity (Hayashi et al., 2008;
Hernandez-Corona et al., 2002; Shimizu,
1996)
12. Asthma (Senevirathne and Kim, 2011).
Diatoms are a kind of microalgae and consist
of biosilica; they are used for drug delivery due
to their pore size and drug-holding capacity.
Isolation, culture and characterization methods
of diatom are important to use in commercial
applications (drug delivery applications).
Diatom biosilica have been used to load several
anti-inflammatory drugs and release them at a
sustainable rate (Dolatabadi and de la Guardia,
2011; Gordon et al., 2009; Losic et al., 2010; Nassif
and Livage, 2011).
6
1. MARINE MICROALGAE BIOTECHNOLOGY: PRESENT TRENDS AND FUTURE ADVANCES
3.6 Biofuels
Algae have been widely used for fuel production because of their high photosynthetic efficiency, high biomass production, and fast
growth (Miao et al., 2004). Microalgae contain
proteins, carbohydrates, and lipids; the lipids
can be converted into biodiesel, carbohydrates
into ethanol and H2, and proteins into the raw
material of biofertilizer (Raja et al., 2013). Biofuel
from microalgae can be processed by using
thermochemical and biochemical conversion.
The thermochemical process can be divided in
to gasification, liquefaction, pyrolysis, and direct
combustion; meanwhile, the biochemical process
can be divided into anaerobic digestion, fermentation, and photobiological activity. By using a
gasification process, the biomass produces CH2,
H2, CO2, and ammonia.
3.6.1 Bioethanol
Bioethanol is an alternative biofuel to gasoline
(Dale, 2007; Demirbaş, 2000; Naik et al., 2010),
which can be produced through yeast fermentation of carbohydrates, specifically from sugary
and starchy feedstock, such as sugar cane, sugar
beets, corn, and wheat (Balat, 2009; Naik et al.,
2010). Certain microalgae species are able to
accumulate large quantities of carbohydrate
within their cells. The carbohydrate is usually
stored at the outer layer of cell wall (Chen
et al., 2013). Through hydrolysis reaction, the
carbohydrate can be hydrolyzed into fermentable sugar (e.g., glucose) for subsequent bioethanol production via fermentation process
(Harun et al., 2010).
3.6.2 Biodiesel
Microalgae have a strong capacity to produce
lipids, which can be easily converted to biodiesel. Transesterification using homogeneous
and heterogeneous catalysts and in situ transesterification are possible methods to produce biofuel from microalgae lipids (Lam and Lee, 2012).
Several advantages have been described for the
FIGURE 3
Concept of two-stage hydrogen production by
microalgae (Rashid et al., 2013).
production of biodiesel from microalgae (Chisti,
2007; Li et al., 2008; Schenk et al., 2008).
3.6.3 Biohydrogen
Biohydrogen from microalgae is an alternative source of energy. A two-stage method of
production has been used: carbon fixation and
anaerobic digestion (Rashid et al., 2013). This
concept of a two-stage hydrogen production by
microalgae is shown in Figure 3.
3.7 Wastewater Treatment
Microalgae-based
wastewater
treatment
relies on the ability of phototrophic microorganisms to supply oxygen to aerobic organic
pollutant degraders and enhance the removal of
nutrients and pathogens. Conventional methods
for the removal of heavy metals from wastewater
include chemical precipitation, coagulation, ion
exchange, membrane processing, electrochemical
techniques, adsorption on activated carbon, etc.
(Dabrowski et al., 2004; Gautam et al., 2014;
Llanos et al., 2010; Wan Ngah and Hanafiah,
2008).
REFERENCES
4. CONCLUSIONS
Microalgae are microscopic plants that
contain potential bioactive materials in the
form of proteins, lipids, glycerols, carotenes,
and vitamins (Avagyan, 2008; Priyadarshani
and Rath, 2012). Potential bioactive metabolites
from microalgae can play a vital role in human
health and nutrition. The designing of new functional foods, nutraceuticals, and pharmaceuticals from marine microalgae makes them one
of the most valuable marine sources. The use of
transgenic microalgae for commercial applications has not yet been reported but holds significant promise (Spolaore et al., 2006). Drug
development is the most promising aspect of
microalgal biotechnology, although screening
of possibilities remains limited (Tramper et al.,
2003). Finally, marine microalgae promise to be
an important alternative as a future bioenergy
source.
Acknowledgments
This research was supported by a grant from the Marine
Bioprocess Research Center of the Marine Biotechnology
Program, funded by the Ministry of Oceans and Fisheries,
Republic of Korea.
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