Detergents Manufacturing Process

Detergents are manufactured using a synthetic surfactant in place of the metal fatty acid salts that
are used in soaps. Made in powder detergents, these detergents are sold as laundry powders, hard
surface cleansers, dishwashing detergents, fabric conditioners etc. Most of the powder detergents
have soap in their mixture of ingredients, however it generally functions more as a foam
depressant than as a surfactant.

Chemical Processes
Powder detergents are manufactured using various processes, such as spray drying,
agglomeration, dry mixing or a combination of these. A brief description of these different
processes is given below -

Spray Drying Process

The different stages / operations performed in a spray drying process, are -

 Dry and liquid ingredients are first combined into a slurry, or thick suspension, in a tank
known as crutcher.

 The slurry is heated and then pumped to the top of a tower where it is sprayed through
nozzles (under high pressure) to create small forming hollow granules as they dry.
droplets. The droplets fall through a current of hot air, thereby
 Collected from the bottom of the spray tower, the dried granules are screened to obtain a
relatively standard size.

After the granules have been cooled, heat sensitive ingredients, which are not compatible with
the spray drying temperatures (like bleach, enzymes and fragrance) are added.
Traditional spray drying process produces relatively low-density detergent powders.
Advancements in technology have enabled the soap and detergent manufacturers to reduce the
air inside the granules during spray drying to obtain higher densities. The high-density detergent
powders can be packed in much smaller packages than those needed previously.

Agglomeration
Agglomeration is detergent powder manufacturing chemical technique that results in high-
density powders. The process involves blending of dry raw materials with liquid ingredients. The
technique involves machines, such as a liquid binder, rolling or shear mixing that causes the
ingredients to collide and adhere to each other, producing larger particles.

Dry Mixing
Dry mixing is a detergent powder manufacturing technique, which is used to blend dry raw
materials. The technique may also involve the addition of small quantities of a liquid.

Ingredients
Mentioned in the table below are the ingredients of a detergent base powder -
Solids
Ingredient Function
Sodium tripolyphsophate(STP) Water softener, pH buffers (to reduce alkalinity).
Sodium sulphate Bulking and free-flowing agent.
Soap noodles Causes rapid foam collapse during rinsing.
Water softener (absorbs Ca 2+ and Mg 2+) in countries where
Zeolite STP is not used; granulating agent for concentrated
detergents.
Increases the negative charge on cellulosic fibers, like cotton
Sodium carboxymethyl cellulose and rayon, causing them to repel dirt particles (which are
positively charged).
Liquids
Ingredient Function
Linear alkylbenzene sulphonic
Surfactant - the main active ingredient
acid (LAS)
Caustic soda solution Neutralizes the LAS.
Coconut diethanolamide or a fatty
Nonionic detergent and foam former.
alcohol ethoxylate
Absorbs UV light and emits blue light, causing aging cotton
Fluorescer
to appear white rather than yellow.
Water Dissolves the various ingredients, causing them to mix better.

Environmental Implications
Detergent powder manufacturing has some specific environmental issues that are not associated
with any other industry. These issues include dust control and volatile organic emissions. Dust
present during the process of delivery and transfer of bulk powdered detergent is a potential
problem. In order to filter out most of the dust, dry and wet cyclones are used, and all the
emissions are monitored.

If the dust level in these does exceeds the acceptable limits, appropriate remedial action must be
taken. The permissible dust levels in emissions are under below 50-mg m -3. The spray-drying
tower also releases volatile organics, which can be minimized by having tight specifications
specifying what can be added as a primary detergent active material. Spot checks must be made
on the total hydrocarbon content of the exhaust gases using a flame ionization detector.

MANUFACTURING PROCESS FLOW SOLUTIONS

Flow solutions in soap and detergents manufacturing refer to the set of equipment, including
pumps and compressors that are used to control the different raw materials movement and flow
control activities. These solutions are formulated to move natural and synthetic raw material feed
stocks from the facilities of petrochemical and oleo chemical processing to transport and storage,
then throughout the blending, filling and bottling operations within the detergent production
facility or the batch or continuous process at the soap plant.

From vegetable or fish oils and glycerin to animal fats and alkaline solutions, like caustic soda and
potash or surfactants to builders, additives, enzymes, fragrances, and brightening agents to fillers,
these pumps and compressors are specifically designed to satisfy the rigorous requirements of
modern syThe various equipment used to control and facilitate the flow solutions of detergents
and soap manufacturing are -

 Vane Pumps
o SNP Sliding Vane Pumps
o SMVP Seal-Less Mag-Drive Vane Pumps
o Motor Speed Vane Pumps
 Eccentric Disc Pumps
 Centrifugal Pumps
 Peristaltic Hose Pumps
 Gas Compressors
 Rotary Vane Compressor

nthetic detergent and soap manufacturing.

SOAPS & DETERGENTS: MANUFACTURING

Soap and detergent manufacturing consists of a broad range of processing and packaging
operations. The size and complexity of these operations vary from small plants employing a few
people to those with several hundred workers. Products range from large-volume types like
laundry detergents that are used on a regular basis to lower-volume specialties for less frequent
cleaning needs.

Cleaning products come in three principal forms: bars, powders and liquids. Some liquid
products are so viscous that they are gels. The first step in manufacturing all three forms is the
selection of raw materials. Raw materials are chosen according to many criteria, including their
human and environmental safety, cost, compatibility with other ingredients, and the form and
performance characteristics of the finished product. While actual production processes may vary
from manufacturer to manufacturer, there are steps which are common to all products of a
similar form.

Let's start by looking at bar soap manufacturing and then we'll review the processes used to make
powder and liquid detergents.

Traditional bar soaps are made from fats and oils or their fatty acids which are reacted with
inorganic water-soluble bases. The main sources of fats are beef and mutton tallow, while palm,
coconut and palm kernel oils are the principal oils used in soapmaking. The raw materials may
be pretreated to remove impurities and to achieve the color, odor and performance features
desired in the finished bar. The chemical processes for making soap, i.e., saponification of fats
and oils and neutralization of fatty acids, are described in the Chemistry section.

Soap was made by the batch kettle boiling method until shortly after World War II, when
continuous processes were developed. Continuous processes are preferred today because of their
flexibility, speed and economics.
Both continuous and batch processes produce soap in liquid form, called neat soap, and a
valuable by-product, glycerine (1). The glycerine is recovered by chemical treatment, followed
by evaporation and refining. Refined glycerine is an important industrial material used in foods,
cosmetics, drugs and many other products.

The next processing step after saponification or neutralization is drying. Vacuum spray drying is
used to convert the neat soap into dry soap pellets (2). The moisture content of the pellets will
vary depending on the desired properties of the soap bar.

In the final processing step, the dry soap pellets pass through a bar soap finishing line. The first
unit in the line is a mixer, called an amalgamator, in which the soap pellets are blended together
with fragrance, colorants and all other ingredients (3). The mixture is then homogenized and
refined through rolling mills and refining plodders to achieve thorough blending and a uniform
texture (4). Finally, the mixture is continuously extruded from the plodder, cut into bar-size units
and stamped into its final shape in a soap press (5).

Some of today's bar soaps are called "combo bars," because they get their cleansing action from
a combination of soap and synthetic surfactants. Others, called "syndet bars," feature surfactants
as the main cleansing ingredients. The processing methods for manufacturing the synthetic base
materials for these bars are very different from those used in traditional soapmaking. However,
with some minor modifications, the finishing line equipment is the same.

Powder detergents are produced by spray drying, agglomeration, dry mixing or combinations of
these methods.

In the SPRAY DRYING process, dry and liquid ingredients are first combined into a slurry, or
thick suspension, in a tank called a crutcher (1). The slurry is heated and then pumped to the top
of a tower where it is sprayed through nozzles under high pressure to produce small droplets.
The droplets fall through a current of hot air, forming hollow granules as they dry (2). The dried
granules are collected from the bottom of the spray tower where they are screened to achieve a
relatively uniform size (3).

After the granules have been cooled, heat sensitive ingredients that are not compatible with the
spray drying temperatures (such as bleach, enzymes and fragrance) are added (4). Traditional
spray drying produces relatively low density powders.

New technology has enabled the soap and detergent industry to reduce the air inside the granules
during spray drying to achieve higher densities. The higher density powders can be packed in
much smaller packages than were needed previously.
AGGLOMERATION , which leads to higher density powders, consists of blending dry raw
materials with liquid ingredients. Helped by the presence of a liquid binder, rolling or shear
mixing causes the ingredients to collide and adhere to each other, forming larger particles.

DRY MIXING or dry blending is used to blend dry raw materials. Small quantities of liquids may
also be added.

Both batch and continuous BLENDING PROCESSES are used to manufacture liquid and gel
cleaning products. Stabilizers may be added during manufacturing to ensure the uniformity and
stability of the finished product.

In a typical continuous process, dry and liquid ingredients are added and blended to a uniform
mixture using in-line or static mixers.

Recently, more concentrated liquid products have been introduced. One method of producing
these products uses new high-energy mixing processes in combination with stabilizing agents.

The final step in the manufacture of soaps and detergents is packaging. Bar soaps are either
wrapped or cartoned in single packs or multipacks. Detergents, including household cleaners, are
packaged in cartons, bottles, pouches, bags or cans. The selection of packaging materials and
containers involves considerations of product compatibility and stability, cost, package safety,
solid waste impact, shelf appeal and ease of use.
 1. INTRODUCTION

In recent years, the rapid depletion of fossil fuels, increase in energy demand, global warming,
increase in price of fossil fuels depends on economic and political behaviors increased
orientation to alternative energy sources. In this context, biodiesel that is one of the renewable
alternative energy sources draws attention because of its useful features such as easily
biodegradable and environmentally friendly. However, biodiesel production from oil crops does
not meet the required demand of vehicle fuel, and recently it is not economic and feasible. It
needs to be improved to produce more economically to be able to compete with diesel in the
market. Vegetable oils and crops which biodiesel produced from are a kind of human food
sources and the shortage on food source cause to go up prices and make the biodiesel high-
priced. To meet the requirements, the interest on algae is increased day by day since this
technology has potential to meet global demand [1]. Microalgae have higher productivity per
area and no need for farm field to grow as opposed to oil crops and animal fat. Microalgae use
sunlight to reduce CO2 to biofuels, foods, fertilizers, and valuable products. Furthermore,
microalgae can be used to get different types of biofuels. Using microalgae as fuel source is not a
novel idea but recently the prices of diesel and global warming hit this solution to the top [2].

Microalgae have lots of advantages for biodiesel production over other raw materials such as
crops, waste cooking oils, and so on. Microalgae have short doubling time which is around 12-24
h since they have a simple structure and capable to high photosynthetic efficiency and they
contain much more amount of oil than other oil crops that can be used as oil source for biodiesel
production. Compared with the oil yields from various oil crops such as corn (172 L/ha),
soybean (446 L/ha), canola (1190 L/ha), jatropha (1892 L/ha), coconut (2689 L/ha) and oil palm
(5959 L/ha), oil yield from microalgae is very high as 136900 L/ha and 58700 L/ha for 70% oil
in biomass and 30% oil in biomass, respectively [2-4].

The other significant feature is that algae can grow everywhere and every season in a year since
there are thousands of algae species that have different adaptations and different properties. They
can grow in saltwater, freshwater, lakes, deserts, marginal lands, etc. In addition to biodiesel
production, algae can be also used as feedstock to produce different valuable products such as
fertilizer, energy, neutraceuticals, protein, animal feed etc. The other significant property is that
microalgae can remove some heavy metals, phosphorous, and nitrogen from water during its
growth. Algae also clean up the water. Moreover, microalgae sequester lots of carbon by
photosynthesis. Utilization of carbon dioxide by algae is significantly lowering the risk for
greenhouse gas effects. Lastly, usage of microalgae for biodiesel almost cancels out the carbon
dioxide and sulfur release to atmosphere [5]. These reasons mentioned above are enough to
believe that microalgae can take the place of fossil fuels completely.

There are many of microalgae studies for biodiesel production. Because the most of the scientists
believe that microalgae will take the place of the petroleum diesel, however, algal biodiesel
production is not feasible yet since there is no much commercial or large scale production of
microalgae for biodiesel. That is why most of the works are focused on decreasing the cost of
biodiesel production or make it competitive versus petroleum diesel. Surely, until these
improvements are achieved, algal biodiesel can not be an accurate alternative. The current
problems making biodiesel expensive can be improved with some innovations. The first of all is
about the algae strain which is also first step of algal biodiesel production. The algae strain
should be better than recent ones. There are natural many kinds of algae strains and isolation of
new natural algae strain may help procedure to be cost effective. The algae strain has to have
high lipid productivity and adaptability to new environments. These features let it produce more
and obtain more oil content [6, 7]. As an example, if the flue gas is used as carbon dioxide
source, microalgae have to be adapted for this situation so that it can tolerate the high
concentration of SOx, NOx, and other gases [8]. That will reduce the cost and increase the
biomass growth rate. The other important innovation should focus on cultivation of algae. The
large-scale production is one of the most cost-intense parts. The innovative thinking should show
a tendency to lower the cost of operation and capital for cultivation systems. As it is explained
below, open ponds are the cheapest way but the efficiency of them has to be worked on.
Moreover, the closed photobioreactors (PBR) are also being improved for a cheaper way to
control and lighten the system. Furthermore, microalgae can be fixed in a cultivation system with
an immobilization technique to get higher biomass. The last way to lower the cost is to produce
sub-products from microalgae beyond biodiesel. There are lots of high value products and sub-
products produced from microalgae such as biogas [9, 10], biobutanol, acetone [11], Omega 3 oil
[12], eicosapentaenoic acid [13], livestock feed [14], pharmaceuticals and cosmetics [15, 16].
Especially sub-products can be preferred for economic support of main process.

For example, recovery of methane from microalgae pulp after biodiesel production develops
renewability of conversion of microalgae biomass to biodiesel process as much as it makes the
cost of process and environmental effects less. The microalgae pulps after oil removed contain
significant amounts of protein and carbohydrate that can convert to biogas by anaerobic
fermentation. Conversion of algal waste to biogas by anaerobic fermentation will play a dual role
for renewable energy production and also sustainable development of microalgal biodiesel
industry [17, 18].

Algae can be also used in bioethanol production. Algae are more uniform and continuous than
terrestrial plant, due to lack of functional parts such as root and leaf composition. Their cell walls
made of polysaccharides that can hydrolyze to the sugar. For this reason, microalgae can be used
as carbon source in fermentation process. Ethanol produced by fermentation can be purified for
using as a fuel, CO2 as a nutrient may also be recycled to algae culture to grow microalgae [19,
20].

In this chapter, algae production methods that cover the algae strain and location selection, algae
cultivation, harvesting, oil extraction, and algal biodiesel production processes are presented in
detail with alternatives. New progresses in this area are also explained.

2. ALGAE STRAINS AND PROPERTIES

Algae are simple organisms including chlorophyll. They can be found in seas, soils and lakes
wherever they can use the light for their photosynthesis. There are two types of main algae
groups. The first group is macro algae, which includes green, brown and red algae. The second
group is microalgae as phytoplankton in the coasts, lakes and oceans, which includes diatoms,
dynoflagellates, green and brownish flagellate, and blue-green algae [21].
The classification of algae can be done in many ways since there is a millions of kind. Also there
is no standard on classification so you can see different types of classification. The taxonomic
group of algae can be given as follow: Archaeplastida, Chlorophyta(green algae),
Rhodophyta(red algae), Glaucophyta, Chlorarachniophytes, Euglenids, Heterokonts,
Bacillariophyceae(diatoms), Axodine, Bolidomonas, Eustigmatophyceae, Phaeophyceae(brown
algae), Chrysophyceae(golden algae), Raphidophyceae, Synurophyceae, Xanthophyceae(yellow-
green algae), Cryptophyta, Dinoflagellates, Haptophyta[22].

Algae are the most common wide photosynthetic bacteria ecologically. To grow algae some
parameters such as amount and quality of ingredients, light, pH, turbulence, salinity, and
temperature become prominent. Macro (nitrate, phosphate, silicate) and micro (some metals, B1,
B12 and biotin vitamins) elements are required in the growth of algae. Light intensity has also an
important role, the light demand changes up to microalgae density and type of microalgae. The
other parameter pH is mostly between 7 and 9 for most of algae strains and mostly the optimum
range is 8. 2-8. 7. The last parameter salinity should be between 20-24 ppt. Moreover, nitrogen
also affects the growth of some algae strains as such as green algae [22-25].

2.1. MACROALGAE

Macroalgae are adapted to life in ocean and it is a plant mostly seen on the costal strips. There
are plenty of macro algae types. Algae can be classified as brown, red, and green based on type
of pigments. Recently, several brown algae types have been used in the industry and energy
production as an alternative source to fossil fuels, and green algae is also studied to produce
biodiesel [26].

Brown algae have xanthophyll pigments and fucoxanthin, which results the colour of brown
algae. These substances mask the other pigments [27]. Polysaccharides and higher alcohols are
nutrition reserves of brown algae but the main carbohydrate reserve is laminarin. The cell walls
of brown algae are made of cellulose and alginic acid. Brown algae have a lot of features such
as: Cytotoxic and antitumor activity, Antifungal activity, Anti-inflammatory activity, Antiviral
activity, Protection against herbivorous animals (fish, sea urchins), Antioxidant activity [21, 28,
29]. Composition of brown algae can vary according to species, their location, salinity and
season. According to analysis, brown algae contain about 85% high moisture and 25 % high
sodium carbonate [26].

Green algae contain chlorophyll a and b. Presence of these pigments makes green color of the
green algae. There are a few reports about second metabolites of green algae. [21]. Moisture
content of green algae is higher than brown algae but they have similar sodium carbonate
content. Green algae species can access higher sugar levels and this makes them useful energy
sources. They also have high cellulose content [26]. Green algae have a lot of features such as:
Anti-inflammatory substances, Cytotoxic and immunosuppressive activities, Antibacterial
activity, Antiviral activity, Antifungal activity [30].

Red Algae have phycoerythrin and phycothcyanin pigments that make red color of these algae.
These pigments mask the other pigments. The cell walls of red algae made of cellulose, agar and
carrageenan [27]. There are approximately 8000 red algae species. In comparison of the other
algae species, red algae are considered as the most important active metabolite resource. They
have a lot of features such as: Cytotoxic activities, Antiviral activity, Anti-inflammatory activity,
Antimicrobial activity, Free radical scavenger activity [21, 31].

2.2. MICROALGAE

There are at least 30000 microalgae species in the world. Microalgae are mostly defined as
unicellular photosynthetic cells but some complex associations create larger colonies. This is a
heterogenic group, which contains prokaryotic organisms similar to bacteria and eukaryotic cells
[26, 32]. Microalgae production is concentrated on particular species, which have special
tolerance for extreme conditions in their growth. This situation enables the production in open
ponds and canals. In future, microalgae production will focus on more advanced species for the
demand of energy and pure monocultures which have specific capabilities like production of
carbohydrate, lipid or hydrogen will be cultivated [33]. According to use of algae, biomass of
microalgae has variable chemical composition. They can be rich or balanced composition of
protein, lipid and sugar. Microalgae selection should be made according to desired biofuels.
Microalgae have important lipid content even in the extreme conditions they reach higher lipid
content [26].

Green algae or diatoms are the most used microalgae species for production of alternative energy
derives. Just a handful of these species has commercial importance. This group contains
Chlorella, Spirulina, Dunaliella and Haematococcus. Only Dunaliella is a dominant sea species.
These are usually cultivated for extraction of high value component like pigments or proteins
[26].

Blue-green algae (cyanobacteria) have a lot of common structural features with bacteria. They
are classified as algae because they contain chlorophyll and other components. They have also
nitrogenic components because all of the prokaryote species convert atmospheric nitrogen to
ammonium [21, 34]. Morphologically blue green algae can have filamentous, conical or
unicellular shape. They have a lot of features such as: anticancer and cytotoxic activities,
antibacterial activity, antifungal activity, immunosuppressive activity [21, 35, 36].

Pyrrhophyta (Dinoflagellates) are unicellular organisms, which are classified as primitive algae.
Large amount concentrations of these organisms exist in ocean surface and they cause fish
deaths. Also because of their pigments, dinoflagellates give the water brown to red coloration in
the sea [34, 37]. Particular dinoflagellate species produce toxin in case of consumed by species
such as shellfish. Consumption of contaminated shellfish by humans can cause a lot of health
problems including death [21].

Bacillariophyceae (Diatoms) are the most versatile and frequent family. They are more feasible
for large-scale productions due to short doubling time and easy to grow. Unlike Dinoflagellates
they create less second metabolites [38].

Microalgae are investigated as biodiesel feedstock because of their high photosynthetic

efficiency, their ability to produce lipids. Macroalgae usually don’t contain lipids too much and
they are taken into consideration for the natural sugars and other carbohydrates that they contain.
These contents can be fermented to produce alcohol-based fuels or biogas.

2.3. LIPID CONTENT OF MICROALGAE SPECIES

As the structure of many microalgae species can accumulate significant amounts of lipid and
provide high oil yield. Their average lipid contents can be reached to 77% of dry biomass under
some certain conditions [39]. Table 1 shows lipid content of some microalgae species.

Microalgae Oil content (dry weight %)

Botryococcus braunii 25-75

Chlorella protothecoides 14-57

Crypthecodinium cohnii 20-51

Dunaliella tertiolecta 16-71

Nannochloris sp. 20-56

Neochloris oleoabundans 29-65

Phaeodactylum tricornutum 18-57

Schizochytrium sp. 50-77

Skeletonema coastatum 13-51

TABLE 1.

Lipid content of some microalgae species [15, 39, 40-45].

Also high productivity is very important beside high oil content. As shown in table 1, microalgal
lipid content can reach 77% by weight of dry biomass but it is observed that there can be low
productivity of Botryococcusbraunii, however, Chlorella appears to be a good choice in
biodiesel production, since it has high productivity though lower oil content [39].

Lipid content can be affected by several parameters such as nutrition, environment, cultivation
phases and conditions growth can affect fatty acid composition [32], Fatty acid composition is
important in microalgae selection because it has a significant effect on biodiesel properties. For
example, if unsaturated fatty acid content is high in algal oils and their presence reduces the
efficiency of esterification to produce biodiesel [39].

Value chain stages of biodiesel production from microalgae can be given as algae and site
selection, algae cultivation, harvesting, filtration, dewatering, oil extraction and biodiesel
production [39].
 3. BIODIESEL PRODUCTION FROM MICROALGAE

The selection of species depends on some factors like ability to usage of nutrition or grow under
specific environment conditions. All these parameters should be evaluated for biodiesel
production.

3.1. SELECTION OF ALGAE STRAIN AND LOCATION

To make algal biodiesel cost effective lots of researchers keep going on algae culturing. The
criteria to select location and sources are mentioned below [46]:

 Water sources and demand, salinity, content

 The region information such as topography, geology
 Weather conditions, isolation, evaporation
 Availability of carbon and food resources

The next decision should be on the algae culturing process type. It can be either batch or
continuous process. Depending on microalgae strain, environmental conditions, availability of
nutrition and moreover industrial pollutions the process type has to be selected. The devices and
apparatuses also have to be adjusted for these conditions and nutrients [39].

Algae strains have different contents, different doubling time (the total biomass per time and
volume) and resistance to change in environmental conditions. Biodiesel production directly
depends on the oil content of microalgae and its efficiency. So that, even the process and
culturing systems are selected perfectly, time and other related factors plays an important role
[39].

3.2. METHODS USED FOR ALGAE GROWTH

Not only the microalgae strain is important for efficiency of oil but also growing conditions are
important. There are different ways to grow algae. Each type of microalgae has a different
mechanism which let them to respond different weather and environmental conditions [39, 47].
Different growing conditions affect the microalgae doubling time. There are 4 growing type
basically: phototrophic, heterotrophic, mixotrophic, and photo heterotrophic. All of them will be
explained in detail.

3.2.1. PHOTOTROPHIC GROWTH

Microalgae are mostly thought to be phototrophic since it requires light [48]. Phototrophic
growing method is based on using light and carbon dioxide to produce chemical energy during
photosynthesis. This is the most common way used to grow microalgae. The best advantage of
the process is using carbon dioxide as a carbon source to grow or produce fatty acid. Since
carbon dioxide is only the carbon source, locations close to fabrics and companies could be
selected to procure carbon dioxide. If it is compared to other growing types, phototrophic method
has the lowest contamination risk [49].
 3.2.2. HETEROTOTROPHIC GROWTH

Some microalgae are not able to grow phototrophic conditions but they can grow in dark using
organic carbon as a carbon source like bacteria. If microalgae is using organic carbon these
microalgae are heterotrophic growing algae. Heterotrophic growth has advantages over
phototrophic growth because light is not required. The biggest problem with the phototrophic is
the light penetration when the density of the culture gets higher. In that way one of the biggest
problems is solved with heterotrophic growth. Heterotrophic growth will be more cost effective
compared to phototrophic growth [48]. And this method is said the most practical and promising
way to increase the productivity [50-52]. Also higher oil rates and efficiency can be obtained
when the algae grow heterotrophic, but the contamination risk is much higher compared to
phototrophic [49].

Microalgae uses different organic carbon sources such as glucose, acetate, glycerol, fructose,
sucrose, lactose, galactose, and mannose, especially growth with sugar is more efficient [49].

Mostly the organism growing heterotrophic should have adaptation property to new habitat as
soon as possible since when culturing to new media the lag phase should be too short, and
durability during processing in fermenters and other machines [48].

3.2.3. MIXOTROPHIC GROWTH

Mixotrophic growth is a combination of phototrophic and heterotrophic growth. Mixotrophic

growth is using organic and inorganic carbon and the process requires light because of
photosynthesis. Thus the microalgae have ability to live in both conditions. Microalgae uses
organic compounds and carbon dioxide as a carbon source and the released carbon dioxide are
also captured with the photosynthesis. Although mixotrophic-growing method mostly is not
preferred compared to heterotrophic and phototrophic growth [49], because of other advantages
even so mixotrophic method is applied in some studies. For example; Park et al. found that
biomass and lipid productivities were boosted by mixotrophic cultivation [53]. Bhatnagar et al.
found the mixotrophic growth of some microalgae strains resulted in 3–10 times more biomass
production compared to that obtained under phototrophic growth conditions [54].

3.2.4. PHOTOHETEROTOTROPHIC GROWTH

When microalgae use organic compounds as carbon sources, sometimes it requires light. The
main difference between mixothrophic and photoheterotrophic is that mixotrophic growth using
organic compounds as energy sources, as photoheterotrophic growth requires light as energy
source. This method is mostly used for production of some beneficial metabolites; however, it is
rarely used for biodiesel production [49]. Metabolisms can split into groups due to pH changes.
Chlorella vulgaris, Haematococcus pluvialis, Arthrospira (Spirulina) platensis strains are the
examples for the growth by mixothrophic, phototrophic and heterotrophic methods. Selenastrum
capricornutum and Scenedesmus acutus are able to grow in phototrophic, heterotrophic,
photoheterotrophic conditions [47].
Algae require more than organic carbon, sugar, protein, oil or any carbon sources. Algae cannot
grow without vitamins, salts, or some other nutrients (nitrogen and phosphor). Moreover, there
are lots of parameters has to be controlled during algae growth to maximize and stabilize the
production. Some of these parameters are oxygen rate, carbon dioxide rate, pH, heat, light
intensity and so on. When appropriate weather conditions and enough nutrients are provided
microalgae grow faster. Mostly doubling time is between 3. 5 h and 24 h [39].

As a result, if we compare different methods mentioned above for microalgae

growth;Heterotrophic growth is much better than the others for the application of biodiesel.
These methods can produce more oil than other growing types. However, heterotrophic cultures
may contaminate especially in open pond systems and result in big problems in large-scale
production. Moreover, organic carbon as a carbon source is an expensive raw material and makes
the process cost higher. Phototrophic growth is an easily scalable and mostly uses the carbon
dioxide from exhaust gas for the production of oil. However, the efficiency of the oil is lower
than heterotrophic growth because the biomass doubling time is higher and total biomass rate is
lower at the end. Phototrophic method mostly preferred to set a cost effective system [49].

3.2.5. CONDITIONS FOR GROWTH OF ALGAE

[Link]. LIGHT

The microalgae growing photosynthetically needs light and the light intensity is the most
significant limiting factor. Algae culture systems mostly use both sun and lamp light. Mostly
lamp-lightened algae culture systems uses wider screens to be able to absorb more light from the
system. For photosynthetic production, at least 50 % of the volume of PBR has to get enough
light [55]. Open raceway ponds, plate, plate PBR, Vertical-column PBRs, Internally-illuminated
PBRs, inclined tubular type, horizontal/continuous type, bubble column and air-lift PBRs are the
systems used for photosynthetic algae growth. Plate photo bioreactor is more efficient than
tubular photo bioreactor because the light can penetrate to bottom more in plate design. Recent
works are on closed system photobioreactors to improve the capacity. Some works are done to
increase the capacity; however the light penetration becomes a major problem. Light source for
open ponds is only Sun. That is why the alteration is not possible for raceway ponds. The depth
of the pond that the only thing can be changed. Thus mostly researches are going on closed
systems to optimize light emission. Mostly photobioreactors in lab scale are lightened by
fluorescence lights from inside and outside [56]. The light wavelength should be between 600-
700 nm to maximize the photosynthesis. Light intensity depends on microalgae density. Higher
algae density requires higher light intensity. Light also affects the lipid content. Yeesang and
Cheirsilp reported that the lipid contents in all strains increased with increasing light intensity in
their study [57].

Changes in light intensity and quality can alter biofuel quality [58]. Each type of microalgae has
its own optimal light absorbing point. If this point exceeds the optimum point, microalgae light
absorption ratio decreases. After a specific point, light decreases the biomass production and this
is called photoinhibition. Photoinhibition processes depend on time and after stress of light for a
few minute biomass loss starts. 10-20 min later more than 50 % damage can be seen. Cheirsilp
and Torpee investigated the effect of light intensity on growth and lipid content of marine
Chlorella sp. and Nannochloropsis sp. The growth of marine Chlorella sp. increased when the
light intensity was increased from 2000 to 8000 lux. But up to 10000 lux its growth decreased.
They reported that this could be some extent of effect from photoinhibition. The growth of
Nannochloropsis sp. continuously increased up to the maximum level when increasing light
intensity up to a maximum light intensity of 10000 lux. [59]. High light intensity limited algal
growth, but gave the benefit of higher lipid content and yield. It can be seen in Ruangsomboon’s
study whose cultures exposed to low light intensity showed a higher biomass compared to others
[60].

To increase the microalgae production, photoinhibition should be cut off or exceed to high light
intense. In addition, photorespiration decreases the photosynthetic efficiency. Therefore the
process has to avoid photorespiration. Photorespiration occurs when the oxygen concentration
increases depending on carbon dioxide [56].

Sara et al. investigated the light effects on microalgae. The research was done by using red and
blue lasers as light source for photosynthetic growth of green algae. The results showed that the
both blue and red lasers increased the algae cell count [61].

Allen and Arnon tested the effect of light on green algae growth. The light intensity was around
16000 lux. There were two samples. One of the samples was analyzed under 11 h darkness and
13 h light. The other sample was analyzed under light for 24 h and the results showed that the
growth rate was same. However after 5 days the growth rate for the sample with 24 h light was
declined [62].

The effects of light on Parietochloris incisa was analyzed by Solovchenko et al. The results
showed that best growth was seen on high light (400 μmol photons m− 2 s−1). With high light
condition, total fatty acid and arachidonic amount was increased due to increase in biomass [63].

Another study (Yeh et al. ) was focused on effects of different light sources on microalgae (C.
vulgaris) growth. In the sutdy, three different light sources was used which are tungsten lamp,
fluorescent lamp (TL5), fluorescent lamp (helix lamp). The results showed that fluorescence
lamps were much better for algae growth. In an other study by Floreto et al., it was mentioned
that high light intensity increased the palmitic acid and most fatty acids ratio [64].

[Link]. CARBON DIOXIDE

Carbon dioxide is the natural carbon source of the microalgae culture. Oxygen is releasing
depending on decreasing carbon amount and it is delivered to the medium. Carbon dioxide is an
general carbon source for photosynthetic microalgae. When the carbon amounts get low, oxygen
is produced by photolysis of water and released to media. Since algae lives in high carbon
dioxide concentration, greenhouse gases, nitrogen dioxide and atmospheric pollutants came from
different sources became a food for algae. The exhausted gases can feed algae production
facilities from fossil fuels and also its efficiency would be increased. Works on usage of stack
gases as carbon source were done but the toxicity of the stack gas components couldn't be
documented well. The amount of carbon dioxide required for the growth relates to type of
microalgae and photo bioreactor. Some types of algae strains are able to keep growing in high
carbon dioxide conditions, in contrast for faster growth lower carbon dioxide concentration is
required [56]. Widjaja studied the effect of CO2 on growth and it was seen that this effect
correlates directly to the lipid productivity since growth was enhanced tremendously by
increasing the CO2 concentration [65]. CO2 requirement can change up to strains. VirthieBhola et
al. reported in their studies that at 15% CO2 concentration there is a 3-fold decline in biomass
yield when compared to the yield produced at a 4% CO2 concentration. This suggests that the
strain under study could not endure CO2 concentrations greater than 4% [66]. Also
Ebrahimzadeh et al. reported that increasing CO2 injection had a significant effect on microalgae
growth [67]. CO2 input is also important. Sonnekus reported that the CO2 should make up 0. 2 -
5% of the total gas flow and being careful about the CO2 input does not lower the pH of the
culture [68].

[Link]. HEAT

Algal growth is also dependent on temperature. For maximum growth there is a need to know the
optimal temperature. The temperature changes also lipid production and composition [69]. The
degree of unsaturation of algal membrane lipids increases if cultures are maintained at
temperatures below their optimum [70]. Other than this temperature is significant for
dissolubility of carbon particles, which helps carbon to be used for photosynthesis. Heat effects
respiration and photorespiration more than photosynthesis. However, if carbon dioxide and light
are the limiting factor, the effect of heat is not significant anymore. Optimal temperature for
microalgae cultures is between 20-24 °C. This can be different according to media composition,
type of culture and strain. The most general cultured microalgae can tolerate the temperature
between 16- 27 °C. The temperatures lower than 16 °C will increase the duplication time and
higher than 35 °C will have a fatal effect on algae [56]. However, these ranges can be changed
by environmental factors such as salinity, pH, carbon dioxide etc.

In the study of Floreta etal., the factors affecting algae growth were determined. Temperature
effect was determined with salinity simultaneously. The results showed that low temperature (15
°C) with high salinity is the best choice. Low temperature increases the level of oleic and linoleic
fatty acids. Moreover, high salinity increases the amount of C16 and C18 poly-unsaturated fatty
acids [71].

[Link]. PH

Microalgae require different pH values according to the media. During high pH concentration,
the carbon dioxide might be limiting factor for growth and photosynthesis. The most used pH
range for algal growth is around 7-9. The optimal pH for algae is between 8. 2- 8. 7. But it can
change with different strains. For example, Weissel and Stadler studied with Cryptomonas sp.
which showed positive population growth rates over a wide pH range, from 4. 4 to 9. 65 [72].
Appropriate pH can be adjusted by ventilation or gassing. There is a complex relationship
between CO2 concentration and pH in microalgal bioreactor systems, owing to the underlying
chemical equilibrium among such chemical species as CO2, H2CO3, HCO3 and CO3. Increasing
CO2 concentrations can increase biomass productivity, but will also decrease pH and this causes
important effect upon microalgal physiology [73]. Water contaminated with a high pH has
negative effects on algal abundance [74]. If there is not enough CO2 gas supply, algae will utilize
carbonate to maintain its growth [75].

Although high concentration of carbon dioxide provides high biomass efficiency, on the other
side higher contamination risk and effect of low pH on microalgae physiology occurs [56].

Except the parameters mentioned above; there are also some parameters which affect on algal
growth or lipid accumulation. Nitrogen, phosphorus and salinity can be examples for these
parameters [76]. Widjaja et al. studied about nitrogen starvation effect on lipid accumulation.
They reported that longer time of nitrogen starvation obviously resulted in higher accumulation
of lipid inside the cells. Under all CO2 concentrations, the lipid content tend to increase when the
algae was exposed to nitrogen starvation condition that total lipid content was higher than lipid
obtained during normal nutrition [75]. Ruangsomboon found the highest biomass concentration
was found under the highest phosphorus concentration [60]. Li Xinet all. have reported in their
study that lipid productivity was not at its highest when the lipid content was highest under
nitrogen or phosphorus limitation [77]. Yeesang and Cheirsilp also studied about nitrogen and
salinity effect. They found an increase in algal biomass under nitrogen-rich condition for all
strains and in the absence of a nitrogen source, no growth was observed. They reported that
although some loss in algal biomass was found, the lipid contents of four strains increased. They
also noticed that growth and lipid accumulation by these microalgae could be affected by
salinity. Under nitrogen-rich condition, all strains survived at high salinity but growth of some
strains decreased [57, 78].

3.3. MICROALGAE CULTIVATION SYSTEMS

Cultivating microalgae can be achieved in open systems like lakes and ponds and in high
controlled closed systems called photobioreactor. A bioreactor is defined as a system, which
carries out biological conversion. Photobioreactors are reactors, which used for prototroph to
grow inside or photo biological reactions to occur [79].

3.3.1. OPEN PONDS

Generally open ponds are used in microalgae cultivation. Open ponds have various shapes and
forms and certain advantages and disadvantages. In the scientific investigations and industrial
applications, raceway ponds, shallow big ponds, circular ponds tanks and closed ponds are used
[80]. Area where pool exist is critical factor for selection of pond type. Ponds become local
climate function due to lack of control in open ponds [80, 81]. Therefore, area contributes to the
success. Open ponds are limited by key growth parameters, which include light intensity,
temperature, pH and dissolved O2 concentration. Another problem seen in open ponds is
contamination. It limits cultivation system of algae, which can grow under certain conditions
[79].

Cost of cultivation systems is an important factor for comparison of open and closed systems.
Construction, operation and maintaining costs are less than photobioreactors in ponds and these
systems are simpler than the others [79, 82].
 3.3.2. PHOTOBIOREACTORS

Nowadays researches are made for designing photobioreactors due to cultivating microalgae.
Photobioreactors offer better control than open systems [2]. Their controlled environment allows
high yield for cultivating.

Productivity is the most important indicator for bioreactor technology. It is very difficult to
compare productivity of bioreactors due to various strains and scale of microalgae [80].

Photobioreactors basically can be tubular and flat type. When it is compared with the other
bioreactors, tubular reactors considered as more suitable for open cultivating. Large illumination
surface of reactor, which made of transparent tubes, is the main factor to being suitable for
cultivation. Tubes can be adjusted in various types, adjustments convenience is depend to the
specification of system.

A general configuration includes straight line and coiling tubes [83]. Reactor geometry is also
important, tubular reactors can be vertical, horizontal or inclined shape. There are important
differences between configurations of vertical and horizontal. Vertical designs provide more
mass transfer and reduce energy consumption; horizontal designs can be scaled but needs more
space. There are more studies about tubular photobioreactors but usually flat type
photobioreactors is preferred because it can offer high cell density [84]. In addition, this type of
reactors is advantageous due to low energy consumption and high mass transfer capacity,
reduction of oxygen increases, high photosynthetic efficiency, no dark volumes when compared
with the other photobioreactors. Suitable reactor design should be provided with maximum cell
mass. Various flat-plate photobioreactor designs are made of glass, thick transparent PVC
materials and V-shape and inclined. Although the other designs are cheap and easy to construct,
glass and PVC is more transparent for maximum light penetration [80, 84-86].

[Link]. FLAT-PLATE PHOTOBIOREACTORS

These systems have large illuminated surfaces. Generally these photobioreactors are made of
transparent materials to utilize the solar light with maximum degree. Dissolved oxygen
concentration is low compared to the horizontal tubular photobioreactors. In this system high
photosynthetic activity can achieve. Although it is very suitable for culturing algae but it has
some limitations [83].

[Link]. TUBULAR PHOTOBIOREACTORS

Most of tubular photobioreactors are made of glass or plastic tubes. They can be horizontal,
serpentine, vertical, near horizontal, conical and inclined photobioreactors. Ventilation and
mixing is generally performed by pump or ventilation systems. Tubular photobioreactor is
suitable with their illuminated surfaces. But one of the important limitations of this system is
poor mass transfer. It is a problem when photobioreactor is scaled. Also photoinhibition is seen
in photobioreactors [83, 87].
If there is not sufficient mixing system cells don’t have enough light for their growth.
Developing mixing systems can provide effective light distribution.

Also controlling culture temperature is very difficult in these systems. Thermostat can be used
but it is expensive and hard to control. Also cells can attach the walls of tubes. Long tubular
photobioreactors are characterized with transfer of oxygen and CO2 [83, 88].

Vertical column photobioreactors are low cost, easily constructed and compact systems. They are
promising for large scale of algae production. Bubble column and airlift photobioreactors can
reach specific growth rate [56].

[Link]. INTERNALLY ILLUMINATED PHOTOBIOREACTORS

Florescent lamps can illuminate some photobioreactors internally. Photobioreactor is equipped

with wheels for mixing algal cultures. Sprayer provides air and CO2 to culture. This type of
photobioreactors can utilize solar light and artificial light [90]. When solar light intensity is low
(night or cloudy day) artificial light is used. Also in some researches, it is told that solar light can
be collected and distributed with optic fibers in cylindrical photobioreactors [91]. Another
advantages of this system are can be sterilized with heat under pressure and minimizing the
contamination [56, 83].

[Link]. PYRAMID PHOTOBIOREACTOR

The Pyramid photobioreactor is using fully controlled and automatic system that increases the
production rate. With this system, it is easy to grow any microalgae at any climate conditions.
The design is in pyramid shape to absorb light more effectively. As mentioned above, light is one
of the significant parameters affecting algae growth rate and with this recent system algae can be
supplied with optimal light intensity. That is why the shape of the system is the last innovation
for production step. So, having optimal light intensity during high microalgae production
decreases the energy consumption. The body design is angled to reduce to pump costs by using
air-lifting method and decrease the deformation on cell walls. Thermo-isolated and high
technologic materials are used to avoid energy lost and over heating [92].

3.4. BIOCOIL MICROALGAE PRODUCTION SYSTEM

Biocoil is a holozoic tubular photobioreactor which made of plastic tubes with small diameter
(between 2.4-5 cm), centrifuges, diaphragm pumps or peristaltic pumping are utilized in this
system. Biocoil design provides equal mixing and reduces the attachment of algae to the walls. It
automates the production process. It is not suitable for all algae species. Some of algae species
damages by circulation system and some of them attach to the internal surface of tubes and
affects algae production negative. In this system, when the level of algae increases maximum
degree, because of the light limitation photosynthesis can slow. Biocoil systems with utilizing
solar light in or outsides can executable. Light is given with an angle so algal cell can utilize
better and photosynthesis occurs easily [89, 93, 94].
 3.4.1. DESIGN OF CULTURE GROWTH SYSTEMS

Depends of local conditions and suitable materials various culture systems can be designed by
various sizes, shapes of construction material, slope and mixing type. These factors affect
performance, cost and resistance. To construct suitable photobioreactor material has main
importance. Materials like plastic or glass relax and rigid shouldn’t be toxic, they should have
mechanical power, resistance, chemical stability and low cost. Tubular photobioreactors are the
most suitable ones for open culture systems. They have big illumination surface, good biomass
productivity and they aren’t expensive because they are made of glass or plastic tubes. Flat-type
photobioreactors are made of transparent materials to utilize solar light energy in maximum
degree. This type of photobioreactors allows good immobilization of algae and they are cleaned
easily [56]. Pond walls and deep side can made of simple sand, clay, brick or cement even PVC,
glass fiber or polyurethane. For coating mostly long lasting plastic membrane is used. (e. g., 1-2
mm thick, UV-resistant, PVC or polyethylene sheets) sometimes to lower the cost uncoating
ponds are used but that time some problems occur like contamination, a layer of mud and sand
[39].

3.4.2. MIXING

Mixing is a process for increasing the productivity of biomass in photobioreactors. Mixing

provides distribution of light intensity, sufficient CO2 transfer and maintains uniform pH. Mixing
is necessary for preventing algae sedimentation and avoiding cell attachment to the reactor wall.
Mixing is also provides equal light and nutrients to all cells and increases the gas transfer
between culture medium and air [95]. The second of priority measures is carbon supply for using
in photosynthesis. In very dense cultures, CO2 from air (includes 0.035 % of CO2) and bubbles
during the culture can be limited for algal growth. CO2 addition creates a buffer for the result of
changing pH in the water [56].

Poor mixing allows cells to clumping like different size of aggregates; therefore it leads 3 phase
(solid-liquid-gas) system in reactor. This situation tends to reduce the mass transfer. But all algae
cannot tolerate agitation. Because they are sensitive to hydrodynamic stress. High mixing rate
can cause the damaging of cells. Mixing in bubble column and air lift reactors can characterize
with axial dispersion coefficient, mixing time, circulation time and Bodenstein number [96].
Analysis of mixing in bubble column shows it has shorter time than airlift reactors. Bubbles
beyond the suction pipe provide less blurry area and causes better exposure to the light. In
addition, existence of suction pipe in airlift reactors causes more effective mixing because
internal loop provides a circulation. Airlift reactor gives information about fluid flow and high
gas-liquid mass transfer rate. Bubble column causes unbalance cell density and these causes to
death of algae [56, 97].

3.4.3. LIGHT PENETRATION

Another key of successfully scale up is light penetration. IIumination in the photobioreactor

affects biomass composition, growth rate and products. Microalgae need light for their
photosynthesis [98]. Photosynthetic active radiation wave changes about 400-700 nm and this is
equal to the visible light [99]. In intense cultures, light gradient changes over the photobioreactor
radius due to the weakening of the light. Reduction of light intensity related to wave length, cell
concentration, photobioreactor geometry and distance of the light transmittance. Light intensity
in photobioreactor related to light way, cell concentration and light which emits by microalgae
[56].

3.4.4. GAS INJECTION

Supplement of CO2 by bubbles is an important factor to be considered in designs. Injection of

CO2 bases on giving CO2 to photobioreactor artificially. Researches show that rich ventilation of
CO2 provides CO2 to algae, supports deooxygenation of suspension, to improve cycling provides
mixing and limits the light inhibition [100]. But high ventilation rate leads to higher cost that is
why in large scale of microalgae production it is not recommended. These researches results for
microalgae production necessary optimum aeration rate of CO2 gas. Includes about 5% or 10%
of CO2 (v/v), rate of 0.025-1 vvm [100]. Volume of air/medium/time is found cost effective for
air mass culture [56].

3.4.5. COMPARISON OF OPEN AND CLOSED CULTURE SYSTEMS

Open and closed culture systems have advantages and disadvantages. Construction and operation
of open culture systems are cheaper and they are more resistant than closed reactors and have
large production capacity [101]. Ponds use more energy to homogenize to nutrients and to utilize
the solar energy for growth their water level cannot be less than 15 cm [41]. Ponds are exposing
to air conditions because water temperature evaporation and illumination cannot be controlled.
They produce large amounts of microalgae but they need larger areas than closed systems and
they are open to other contaminations from the other microalgae and bacteria. Also when
atmosphere has only 0. 03-0.06 % of CO2, mass transfer limitation slows the growth of
microalgae cell.

Photobioreactors are flexible systems, which can operate for biological and physiological
characteristics of cultured microalgae. It can be possible to produce microalgae, which cannot
produce in ponds. Exchange of gas and contaminants between atmosphere and cultured cells in
photobioreactor is limited or blocked by reactor walls [39]. Depends on the shape and design,
photobioreactors have more advantages than open ponds. Culture conditions and growth
parameters can be controlled better, it prevents evaporation, reduces loss of CO2, provides high
microalgae density or cell concentration, high yield, creates more safe and preserved
environment, prevents contamination. Despite the advantages, photobioreactors have problems to
be solved. Over heating, biological pollution, accumulation of oxygen, difficulty of scale-up,
high cost of construction and operation and cell damage because of shear stress and degradation
of material in photo phase are main problems in photobioreactors [39].

Comparing photobioreactors and open ponds is not easy because growth of algae related to al lot
of different factors. Three parameters are considered in algae production units for yield [41]:

 Volumetric productivity (VP): productivity per unit reactor volume (expressed as g/L. d).
 Areal productivity (AP): productivity per unit of ground area occupied by the reactor
(expressed as g/m2d).
  Illuminated surface productivity (ISP): productivity per unit of reactor illuminated
surface area (expressed as g/m2d).

According to researches closed systems don’t provide advantage for areal productivity but
provide volumetric productivity (8 times) and cell concentration (16 times) more than open
ponds [39, 41].

3.4.6. COMPARISON OF BATCH AND CONTINUOUS PROCESS

Photobioreactors can be operated in batch or continuous process. There are a lot of advantages
for using continuous bioreactors than batch bioreactors. Continuous bioreactors provide more
control than batch bioreactors. Growth rates can be regulating in long time periods, can be saved
and with variable dilution rates biomass concentration can be controlled. With steady state
continuous bioreactors results is more dependable, products can be easily produced and can be
reached desired product quality. Continuous reactions offer many opportunities for system
research and analysis [102].

But some type of bioreactors is not suitable for continuous process. For some productions, cell
aggregation and wall growth can inhibit the steady state growth. Another problem is loss of
original product strain in time. Mixtures viscosity and heterogenic nature make difficult for
maintaining filamentous organisms. Long growth periods increase the contamination risks [83].

3.5. HARVESTING ALTERNATIVES

There are several ways to harvest microalgae and dry them. Some main harvesting methods are
sedimentation, flocculation and filtration.

Sedimentation: When a particle moves continuously in a phase, the velocity is affected by two
factors. First of them is increasing the velocity because the density gradient between particle and
fluid create buoyant force. At the end, buoyant force gets equal to dragging force and particle
starts moving with a constant velocity. The same idea is applied to collect microalgae from the
ponds. Gravity force is used for settling of suspended particles in fluid. This method is cheap and
easy. However, the particles suspended in the fluid have to be incompressible. The problem with
the Scenedesmus sp. and Chlorella sp. is that they are compressible. That is why sedimentation
cannot be used for these types [103]. For low value products, sedimentation might be used if it is
improved with flocculation [104].

Flocculation: is also used for harvesting microalgae. The general idea is microalgae carries
negative charge on it and if the flocculants disappear the negative charge, algae starts
coagulation. Some used flocculants are Al2(SO4)3, FeCl3, Fe2(SO4)3 [105].

Filtration: This is one of the most competitive methods for the collection of algae. There are
different types of filtrations, for example, dead end, microfiltration, ultrafiltration, pressure filter
and vacuum filter. Mostly filtrations require the liquid media with algae to come through
filtration. Filter can be fed until a thick layer of microalgae is collected on the screen. This
method is very expensive for especially microalgae. The pore sizes of the filters are the most
important part. If the pore size is bigger than algae you cannot collect it. In contrast, if the pore
size is too small it might result in decrease of the flow rate and block the pores [106].

3.6. EXTRACTION OF LIPID FROM MICROALGAE

There are a lot of methods for extraction of lipid from microalgae but the most common
techniques are oil presses, liquid-liquid extraction (solvent extraction), supercritical fluid
extraction (SFE) and ultrasonic techniques. Oil presses are usually used for extracting of lipids
from nuts and seeds. The same process and devices can be used for lipid extraction from
microalgae. For the purpose of this process to be effective, firstly microalgae must be dried.
Presses use pressure for breaking cells and removing oil [107]. This method can extract 75% of
oil but in longer extraction times it is less effective [80].

Solvent extraction is more successful for extracting lipids from microalgae. In this method
organic solvents such as hexane, acetone, and chloroform are added in the algae paste. Solubility
of oil is higher in organic solvents than water. Therefore solvent breaks the cell wall and extracts
oil easily. Solvent extraction continues with distillation process for separating oil from the
solvent [108]. Hexane is cheap and has high extraction capacity. For this reason it is reported to
be the most effective solvent in extractions.

In addition to this studies, 2 stage process using ethanol improves lipid extraction. The yield of
recovery of oil reaches about 80%. Butanol is also effective in extraction of lysophospholipids.
But evaporation of butanol is difficult and there are some impurities because of its high polarity
[80].

Supercritical extraction uses high pressure and temperature for breaking cells. This method is
widely used and efficient for extraction time. Studies reported that temperature and pressure
don’t affect the yield of components but it affects extraction rate. Similar effects are seen in SFE
system and solvent extraction [109].

Another method is using ultrasonic techniques. In this method microalgae is exposed to high
intensity ultrasonic waves and these waves creates bubbles around the cell. Shock waves are
emitted by collapsing bubbles. It breaks cell wall and desired components release to the solution.
This method is also improves the extraction rate with the same way. This technique is widely
used in laboratory scale but in commercially scale there is not enough information about cost and
applicability [110, 80].

3.7. BIODIESEL PRODUCTION FROM OIL

After extraction there are 4 main methods for producing biodiesel: direct used and mixing with
raw oils; microemulsion; pyrolysis and transesterification.

3.7.1. DILUTION
This is a dilution method that certain proportion of vegetable and waste oils blended with diesel
fuel and another solvent. The most used oils for producing biodiesel with this way are waste oils
and vegetable oils like sunflower and rapeseed.

Direct use or blending generally considered being unsatisfactory and impractical for both direct
and indirect diesel engines. There are specific problems such as high viscosity, acid composition,
free fatty-acid content, gum formation because of oxidation, polymerization during storage and
combustion, carbon deposits and also lubricating-oil thickening [111].

Dilution of vegetable oils with solvents lowers the viscosity. The viscosity of oil can be lowered
by blending with pure ethanol [112]. The low viscosity is good for better performance of engine,
which decreases with increasing the percentage of diesel [33]. In this method there is no
chemical process and viscosity can be lower but there are also carbon deposits and lube pollution
problems to be solved. To solve problems caused by high viscosity, micro-emulsion, pyrolysis
and transesterification methods are used [113].

3.7.2. MICRO-EMULSION

It is defined that the size of 1-150 nm, the two immiscible liquid organic mixtures with ionic or
non-ionic, self-formed stable colloidal distribution. With this method it is possible to form
alternative diesel fuels except petroleum [28]. In this method vegetable oils with an ester and
dispersant (co-solvent), or of vegetable oils, an alcohol and a surfactant, with or without diesel
fuels can be used to make a microemulsion. Due to their alcohol contents, microemulsions have
lower volumetric heating values than diesel fuels. But these alcohols have high latent heats of
vaporization and also tend to cool the combustion chamber, which cause a reduction of nozzle
coking. A microemulsion made of methanol and vegetable oils can perform like diesel fuels
[111]. To solve the problem of the high viscosity of vegetable oils, microemulsions with solvents
and immiscible liquids, such as methanol, ethanol, 1-butanol and ionic or non-ionic amphiphiles
have been studied [114].

3.7.3. PYROLYSIS

Pyrolysis is the conversion of organic substance into another by means of heat or by heat in the
presence of a catalyst. Vegetable oil, animal fat, algae oil, natural fatty acids or methyl esters of
fatty acids can be pyrolyzed [111]. Although this method is not very cheap, however, fuel can be
produced without extraction of lipids or hydrocarbons. More uniform product can be obtained
and ideally increases yields over transesterification with this method [115]. Products are
chemically similar derived from petroleum products, which are to gasoline and diesel fuel
derived [28]. Also with pyrolysis some low value materials and sometimes more gasoline than
diesel fuel are produced [116]. In comparison between pyrolysis and the other cracking
processes, pyrolysis is seen more simple, pollution free and effective [33]. Sharma et al. reported
that pyrolysis of the vegetable oil can produce a product which has high cetane number, low
viscosity, acceptable amounts of sulfur, water and sediments contents, acceptable copper
corrosion values [117].

3.7 4. TRANSESTERIFICATION
Transesterification of the oil is the most promising solution to the high viscosity problem [114].
In this process, triglycerides are converted to diglycerides, then the diglycerides are converted to
monoglycerides, and the monoglycerides are converted to esters (biodiesel) and glycerol (by-
products) [118]. There are three common kinds of catalysts used in transesterification process
such as lipase catalysts, acid catalysts and alkali catalysts. Each catalyst has advantages and
disadvantages [113].

In the acid-catalytic transesterification, the reaction can be catalyzed by sulfuric, phosphoric,

hydrochloric and organic sulfonic acids. Very high yields can be obtained by using this catalyst.
These reactions need the use of high alcohol-to-oil molar ratios in order to obtain good product
yields in practical reaction times. But ester yields do not proportionally increase with molar ratio
and the reaction time is very long (3–48 h) [114, 119, 120]. Xu et al. studied the acidic
transesterification of microalgae (Heterotrophic C. Protothecoides) oil. They used methanol for
alcohol and they achieved 80% of FAME yield [121].

Johnson made a study on Schizochytrium limacinum microalgae species. He converted this algal
oil to biodiesel with acidic transesterification and he achieved 82.6% of biodiesel yield [122].

In the alkali-catalytic transesterification, the reaction can be catalyzed by alkaline metal

alkoxides, and hydroxides, as well as sodium or potassium carbonates. Sodium methoxide is the
most widely used biodiesel catalyst. This reaction is faster than acid-catalytic transesterification
and reactions can occur in low temperatures with a small amount for catalyst and with little or no
darkening of colour of the oil [114]. High quality can be obtained however this process is very
sensitive to the presence of water and free fatty acids and needs lots of methanol. If the raw
materials have a high percentage of free fatty acids or water, the alkali catalyst reacts with the
free fatty acids to form soaps [113]. There are some studies on microalgae oil to produce
biodiesel by using alkali transesterification. Velasquez-Orta et al. studied on biodiesel production
from Chlorella vulgaris. In that study, alkali transesterification was used for conversion and they
achieved 71% of FAME yield [123]. Ferrentino et al. studied on biodiesel production from
microalgae too. They used Chlorella sp. oil and their production method was alkali
transesterification. They have obtained high yield from their experiment [124]. In another study,
Carvalho et al. used alkali transesterification for biodiesel production from algae oil. In their
study, they used Chlorella emersonii oil and they have obtained 93% conversion yield [124].

It can be seen that there are some problems such as recovery of glycerol or removing catalysts
from product and need of wastewater treatment in acid or alkali-catalytic transesterification.
Enzymatic catalysts like lipases are able to catalyze the transesterification of triglycerides
effectively. With this process glycerol can be easily recovered however enzymatic catalysts are
often more expensive than chemical catalysts. The high cost of enzyme production is the main
obstacle to the commercialization of enzyme-catalyzed processes. But using solvent-tolerant
lipases and immobilized lipases can be a solution for this. Lipase-catalyzed transesterification is
considered to be one of the most effective reactions for production of biodiesel [114]. In another
study Tran et al. used microalgae oil (Chlorella vulgaris ESP-31) for producing biodiesel. Their
method was enzyme-catalyzed transesterification and they used lipase in this process. In the
result, they reported that they achieved 94.78 % of FAME yield [126]. Table 2 presents the
transesterification studies for biodiesel production from microalgae oil.
Supercritical process, microwave-assisted method and ultrasonic-assisted process are novel
methods used in biodiesel production area. Since these methods are novel methods and also
algae are new materials for biofuel area, there is a few studies biodiesel production from algae
oil with these novel methods, these studies were reviewed and presented below.

With supercritical process biodiesel production can be easily achieved without catalysts.
Supercritical fluid is a substance whose temperature and pressure is above the critical point.
These fluids are environmentally friendly and economic. Usually water, carbon dioxide and
alcohol is used for supercritical fluid. In biodiesel production generally supercritical methanol
and supercritical ethanol is used. Advantages of this process are being easier for purification,
shorter the reaction time and more effective reaction [130]. In the study of Patil et al., using
supercritical methanol produced biodiesel. The wet algae were used and the ratio of alcohol/ oil
was chosen as 9:1. The temperature of the reaction occurred at 255 and 1200 psi and resulted in
90% of FAME yield [131].

Microwaves activate differences in small degrees of polar molecules and ions, because the
molecular friction and chemical reactions start. Molecules have not the enough time to relax and
heat generation occurs in a short time because energy interacts with molecules very quickly.
Transesterification reaction is carried out with microwave in a short time and microwave results
in an efficient manner. As a result in a short time separation and pure products with high yield is
obtained. Thus, production costs and the formation of by-product are reduced [130]. Patil et al.,
made a study on biodiesel production from dry microalgae by using microwave-assisted process.
KOH was used as catalyst in the study and microwave condition is set to 800 W. The
performance of the study is around 80% [132]. The other study with macroalgae for microwave-
assisted algal biodiesel was showed that methanol to macroalgae ratio of 1:15 was the best
condition. In the study, sodium hydroxide concentration was 2 wt % and reaction time of 3 min
for the best condition [133]. Koberg et al. was reported the study used Nannochloropsis for algal
biodiesel production with microwave-assisted method. The higher biodiesel yield was observed
which was around 37.1% with microwave technique. The same conditions for sonication
technique resulted in lower yield [134].

Alcohol /
Algae strain Method Alcohol oil molar Temp. Time Results Ref.
ratio

Heterotrophic C. 80%
Acidic
Protothecoides Methanol 56:1 30 °C 4h (FAME [121]
trasesterification
(microalga) Yield

Enzymatic 94.78%
Chlorella vulgaris ESP- 25-40
transesterification Methanol 98.81 48 h (FAME [126]
31 (microalga) °C
(Lipase) Yield)

71%
Chlorella vulgaris in situ alkaline 75
Methanol 600:1 60 °C (FAME [123]
(microalga) transesterification min
Yield)
 Alcohol /
Algae strain Method Alcohol oil molar Temp. Time Results Ref.
ratio

97.5%
Nannochloropsis heterogeneous
Methanol 30:1 50 °C 4h (FAME
oculata (microalga) transesterification
Yield)