MARINE Microalgae

potential source for biofuels

INTRODUCTION
⚫ Unicellular species
⚫ Commonly found in marine and freshwater
⚫ Size ranging from a few micrometers to a few
hundreds of micrometers
⚫ Estimated around 2x105 to 8x105 species exist
⚫ Three important classes in terms of
abundance : diatoms (Bacillariophyceae);
green algae (Chlorophyceae) : golden algae
(Chrysophyceae)

Promising source for several bioactive

Polymers, peptides, fatty acids,
carotenoids, toxins, and sterols
 Why microalgae?

1. The microalgae can be produced all year round

2. They can grow almost anywhere
3. The rapid growth
4. Numerous species
5. The algae need less water than terrestrial crops
6. Algae cultivation does not require herbicides or
pesticides application
7. They grow to high densities and have high
productivity compared to typical terrestrial plant
8. They are well-adapted to survive under a large
spectrum of environmental stresses
APPLICATIONS OF MARINE MICROALGAE
 1. Nutraceutical
• 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
hycobiliproteins, which have health-promoting properties such as
vitamin precursors, antioxidants, immune enhancers, and
anti-inflammatory agents.

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.
2. Cosmeceuticals

• 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).
 3. Pharmaceuticals

• 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; oaman et al., 2004;
Thillairajasekar et al., 2009).
4. BIOFUELS

• 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).
Classification of biofuels
4.1. Bioethanol
• is an alternative biofuel to gasoline (Dale, 2007; Demirbas¸, 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).
4.2. Biomethane

• The remaining algae biomass slurry left after fermentation may be

used in the anaerobic digestion process while keeping the pH in the

range of 6-9. This process produced methane, which can further be
converted to produce electricity (Ueda et al., 1996)

• Biogas produced from anaerobic microorganisms by anaerobic

digestion

• Residual biomass from anaerobic digestion can be further

reprocessed to make fertilizers

4.3. Biodiesel

• have a strong capacity to produce lipids, which can be easily

converted to biodiesel.

• Microalgae are a potential raw material for biodiesel production, as

they meet all of the following requirements. They possess a high

growth rate and provide lipid fraction for biodiesel production (Song
et al., 2008).

• Microalgal lipids are mostly neutral lipids with a lower degree of

unsaturation. This makes microalgal lipids a potential replacement

for fossil fuels.
 Applied algae for biodiesel

1. Species and Strain Selection

• An important step in achieving microalgal culture on

a large scale is selection of the best-suited species for
growth.
• Local microalgal species should be collected because
it can be expected that they have a competitive
advantage under the local geographical, climatic, and
ecological conditions.
• Selection is performed based on the considered
application, such as food production, chemicals,
drugs, biofuels, or wastewater treatment
Concerning biofuel production, the evaluated characteristics of microalgae are :
Screening and cultivation steps
1) Energy yields (photoconversion efficiency);

2) Valuable coproducts (hydrocarbons, proteins, lipids, etc.);

3) environmental tolerance (temperature, salinity, and pH);

4) mass culture performance (resistant to predators in open ponds and robust toward
shear stresses in photobioreactors);
5) media requirements (vitamins, trace metals,etc.);

6) data available on related strains (Chisti, 2007; Dillschneider et al., 2013)

harvesting step
cell size and cell wall properties should promote autoflocculation (cheap harvesting process),
and high sinking speed is required
biodiesel production
cell wall properties are important in oil extraction efficiency (Goncalves et al., 2013)
• Fluorescent dyes such as Nile red (9-diethylamino-5H-benzo[a] phenoxazine-5-one) and
BODIPY 505/515 (4,4- difluoro-1,3,5,7-tetramethyl-4-bora- 3a,4a diazas- indacene) are
employed to screen microalgae for lipid accumulation, which can be used for biofuel
production (Govender et al., 2012).
2) Microalgae cultivation
2.1 Nutritional Requirements of Microalgae
• inorganic nutrients such as nitrogen, phosphorus,
and vitamins and trace elements such as iron,
manganese, selenium,zinc, cobalt, and nickel
• The growth and the biomass productivity of any
species depend on the type of nutrient sources, its
quantity, and the relative proportions of C, N, and P
of the culture medium.
• Nutrient sources, especially the nitrogen, such as
nitrate, urea, and ammonium, and carbon sources
• other than carbon sources, nitrogen and phosphorus
in 16:1 ratio are believed to be the best composition
for the proper growth of most of the algal species
(Richmond, 2008).
• The production of lipids, proteins, and carbohydrates is intrinsically linked to
environmental conditions (light, temperature, and pH), nutrient availability, and
presence of other microorganisms (Abdel-Raouf et al., 2012; Munoz and
Guieysse, 2006).
• Nitrogen is an important element for the production of proteins and nucleic
acids (Yen et al., 2014). It accounts for 7-20% of cell dry weight. This element
can be available in the form of nitrate, nitrite, ammonia, or urea. The absence
of this nutrient promotes the accumulation of lipids and reduction of protein
content (Cakmak et al., 2012; Li et al., 2008; Rodolfi et al., 2009).
• Sulfur is a structural component of some amino acids and vitamins and is
important in the production of chloroplasts (Barsanti and Gualtieri, 2006).
• Potassium is important for several enzymes and is involved in protein synthesis
and osmotic regulation (Sydney et al., 2014).
2.2 Cultivation System for Microalgae
Cultivation systems for microalgae are classified into two main
categories: open cultivation and closed photobioreactors (PBRs).
2.2.1 Open Ponds and Raceways
• the oldest cultivation technique and has been used since 1950

• These microalgal cultivation systems may broadly be categorized as natural open

ponds (lakes, lagoons, ponds) and artificial ponds (raceway ponds).
• The major advantages of cultivation of microalgae in open ponds or raceway
ponds are its scalability, simple design, high production capacities, and lower
operating and maintenance costs (Ugwu et al., 2008).
• The disadvantages : microbial contamination such as bacteria, phycophases,
parasites, zooplanktons, and growth of unwanted algal species, evaporative
losses, low diffusion of atmospheric CO2, requirement of large land area, poor
productivity, limitation to a few strains, as well as extreme weather conditions
(rain, temperature, and light intensities) are the major disadvantages of open
pond systems (Ugwu et al., 2008).
• Closed raceways are less vulnerable to contamination and environmental
disturbances in comparison to open raceway ponds.
2.2.2 Photobioreactors
• PBRs are made up of glass/Plexiglas/transparent PVC material with
internal or external illumination and provided with a controlled gas
exchange and mixing/circulation of media.
• The size and type of these PBRs may vary with strain type and end use of
biomass and scalability
• The major algal growth limiting factors such as light type, intensity,
duration, temperature, mixing and gas exchange, CO2 and evaporation
loss, etc. can be easily controlled and managed in such reactors.
• less prone to bacteria and other contamination

• it can be used indoors or outdoors and is customizable according to the

specific requirements and growth conditions.
• BUT, The major limitations with these systems are the economics and
huge capital cost involved in installation and maintenance.
 Main configurations of photobioreactors: (a) air-lift
bioreactor (van Benthum et al., 1999); (b) tubular
bioreactor (from http://www.omega3.company); and
(c) flat-plate bioreactor. (From
http://www.nanovoltaics.com.)

• Various types such as flat panel (flat plate), vertical/inclined tubular, helical,
airlift, horizontal/serpentine tubular airlift, bubble column, membrane or hybrid type
PBRs (Qiang and Richmond, 1996; Rubio et al., 1999; Ugwu et al., 2008).

• In all PBR typologies, flat-plate bioreactors are the ones that can achieve higher
cell densities, presenting additional advantages: (1) lower power consumption; (2) high mass
transfer rates between gaseous and liquid phases; (3) absence of dark volumes; and (4) high
photosynthetic efficiency. The main drawbacks of PBRs are the investment and operational costs,
reasons why the most current microalgal biomass production is performed in open ponds (Posten,
2009).
 2.2.3 Factors Limiting Microalgal Growth

• a substantial increase of the salt concentration in growth medium beyond the

threshold of any species leads to inhibition of photosynthesis and growth; in
extreme conditions, plasmolysis or cell bursting can occur (Fogg, 2001; Kirst,
1990).
• Higher intensities damage the light receptors of the chloroplasts, which results
in photoinhibition and thus reduced biomass production (Lee, 1999).
3. Harvesting and Processing
3.1 Harvesting techniques
Microalgal harvesting is the removal of biomass from the
culture medium. The selection of adequate harvesting
methods depends on the characteristics of the target
microorganism and also the value of the products that could
be commercialized from the biomass.
 1

3
Conventional methods are centrifugation, flocculation, and filtration.

• Chemical coagulation/flocculation is a low-cost harvesting method.

Coagulation/flocculation is usually followed by gravity sedimentation.

While coagulation involves pH adjustment or electrolyte addition, flocculation is
based on the addition of cationic polymers (Banerjee et al., 2013; Show and Lee,
2014).

• Filtration, it is more effective only for small volumes. For volumes greater than

20 m3 d1, centrifugation may be more economical (Grima et al., 2003; Rossignol et

al., 1999).

• Centrifugation is the fastest harvesting method and can be applied to the great

majority of microalgae. However, it is also the most expensive due to high energy
requirements. Thus, this process is only applied for high-valued products (Grima et
al., 2003; Lee et al., 2009; Sim et al., 1988).
3.2 Drying Techniques

• to remove moisture content (to 12% or less) to avoid interference with solvents

used in downstream processes.

• It can be performed using dryers (spray drying, freeze dryingdlyophilization, and

fluidized bed drying) or by exposing the biomass to solar radiation (Brink and
Marx, 2013).

• Sun-drying corresponds to lower production costs as well as power consumption.

However, this process may not be efficient due to the high water content in
biomass.

• Spray drying is the most commonly applied method

4. Oil extraction

• mechanical crushing followed by solvent extraction

• osmotic shock or ultrasonic treatment to break the

cells

• autoclaving, beadbeating, microwaves, sonication,

and 10% NaCl solution

• Recent research of bio-oil production by pyrolysis of

biomass has received much interest (Huang et al.,

2010; Miao and Wu, 2004). more suitable for
microalgae because of the lower temperature required
for pyrolysis and the higher quality oils obtained
(Bridgwater et al., 1999).
5. Biodiesel production

• Microalgae contain proteins, carbohydrates, and lipids.

• Lipids can be converted into biodiesel, carbohydrates to ethanol and H2, and proteins as

raw material of biofertilizer. Lipid is a general name for plants and animal products that
are structurally esters of higher FAs. The FAs are a variety of monobasic acids such as
palmatic (C16:0), stearic (C18:0), oleic (C18:1) (Klass, 1998), linoleic (C18:2), and
linolineic (C18:3). The FAs can be classified as medium chain (C10eC14), long chain
(C16eC18) and very long chain (>C20) species and FA derivates.

• Lipids can be divided into two main groups: (1) the storage lipid (neutral or nonpolar

lipid) and (2) structural (membrane or polar lipid). For biodiesel production, neutral
lipids have to be extracted (Alcaine, 2010).

• The group of neutral lipids is formed by triacylglycerols or TAGs, steryl esters, and wax

esters (Lang, 2007). TAGs generally serve as energy storage in microalgae that once
extracted can be converted into biodiesel through transesterification reactions.
Transesterification and/or Esterification

• Different catalyst and amount of FFA

PROPERTIES OF MICROALGAE DERIVED
BIODIESEL
• The most common FA profile of algae consists mainly of palmatic

(C16:0), stearic (C18:0), oleic (C18:1) (Klass, 1998), linoleic (C18:2),

and linolineic acids (C18:3) (Knothe, 2009).

• The minor FAs are C8:0, C10:0, C12:0, C13:0, C14:0, C15:0, C15:1,

C16:1, C18:3, C18:4, C20:0, C20:2, C20:5, C20:6, C22:6, C24:0, and
C24:1.

• Microalgal lipids are predominantly polyunsaturated (C16:2, C18:2,

C20:2, C16:3, C18:3, C20:3) and therefore they are more prone to
oxidation. (This is a serious issue with biodiesel while in storage.)
• A good-quality biodiesel should have a 5: 4:1 mass FA ratio of C16:1, C18:1,
and C14:0, as recommended by Schenk et al. (2008). Of the nine
microalgae species investigated by Islam et al. (2013b), the fatty acid
methyl ester (FAME) composition of Nannochlopsis oculata was closest to
the recommended ratio of 5.1:3.5:1, but eicosapentaenoic acid (EPA) is
also present in appreciable quantities (fourth most dominant FA).
• From eight marine water microalgae species investigated by Barman et al.
(2012), the FAME composition of Phoramidium tennue was closest to the
recommended ratio of 5: 3.16: 1
• According to Knothe (2008), the ideal biodiesel feedstock would be
composed entirely of C16:1 and C18:1 (monounsaturated FAs).
• In practice, a biodiesel feedstock should have high concentrations of C16:1
and C18:1 with less variation in the FA profile.