Table 1: Advantages and Disadvantages of Biodiesel Oil That Are Produced From Vegetables Oils

TOWARDS CHEAPER MICROBIAL OIL FOR BIODIESEL PRODUCTION November 29, 2013
1.0 Abstract
Biodiesel, an ideal alternative diesel, is generally produced by transesterification. With
the rapid expansion of biodiesel, microbial oil might be one of the oil feedstock for
biodiesel production. Many microorganisms are able to accumulate oils under some
special cultivation conditions. Compared to the vegetable oils, microbial oils have many
advantages. However, the high production cost has become the major barrier to the
production of microbial oil. This term paper is to discuss the cheaper microbial oil for
biodiesel production. So, a cost-effective microbial oil will be available in market one
day.
2.0 Introduction
2.1 Biodiesel
Biodiesel produced from vegetable oils or animal fats which consist of long-chain
alkyl esters are believed to be an ideal replacement of diesel. It is made through a
chemical process which converts oils and fats of natural origin into fatty acid
methyl esters (FAME). The process of converting the vegetable oils or animal fats
into biodiesel is known as transesterification. The largest possible source of
suitable oil is produced from vegetables oils such as Soybeans or Rapeseed. But,
most of it is produced from waste vegetable oil sourced from restaurant, chip
shops and food industries, which are treated to remove impurities.
Table 1: Advantages and disadvantages of biodiesel oil that are produced from vegetables oils.
ADVANTAGES DISADVANTAGES
can be described as ‘carbon neutral’ as the fuel increase in price of food-grade oils and food
produced no net output of carbon that is in the shortage besides increase in the cost of
form of carbon dioxide (CO2). biodiesel and preventing its usage
has lower toxicity drought happened during the dry climates

 help in increasing the people health  plenty of water may need for the
performances plantation
 lack of sulfur in 100% biodiesel besides vast area is needed for the growth of
extends the lifespan of the catalytic the plants
WONG WIN YEE, LIM WEI MEI, KONG KA MAN, LEE LYE QING, LAW LI CHIN Page 1
converters.  deforestation
 soil erosion
able to blend in with other energy resources biodiesel fuel starts to become cloudy and
and oil begin to form wax crystals during low
 can be used in existing oil heating temperature
systems and engines without making  making it unable to pump
any alterations.  can clog fuel lines and filters in a
vehicle’s fuel system
has a higher flash point has a higher nitrogen oxide emission
 safer in the event of crash  On average pure biodiesel has about
 safer to handle compared to the fossil 10% higher nitrogen oxide (NOX)
fuels. emission than conventional petroleum
diesel
 NOX plays a role in the localized the
formation of smog and ozone
 When NOX dissolves in atmospheric
moisture it can cause acid rain
lubricating properties of the biodiesel degrade and soften the rubber hoses in some
 aid in lengthening the lifespan of the old engines and dirt is collected in the fuel
engines filter and thus clogging it
Biodegradable controversy concern about ‘we should plant

food to feed the hungry child, instead of using
it to power our trucks’
2.2 Microbial oil
Microbial oil is known as the single cell oils. They are triglycerides that are
effectively produced through the oil accumulation in the oleaginous
microorganisms, such as yeast, fungi, bacteria, and microalgae under some special
cultivation conditions with the help of light, carbon dioxide, waste water and
lignocelluloses biomass which act s the feedstock for microbial oil production.
But, they are mostly produced by microalgae. Microalgae are made out of
unicellular structure, which makes their entire surface available for light capture
and mass transfer. This leads to be high photosynthetic efficiency and allows a
potentially higher oil yield per area compared to the best oilseed crop. Microalgae
are generally autotrophic eukaryotic cells, they can grow autographically and/or
heterotrohically, with a wide range of tolerance to different temperature, salinity,
pH and nutrient availabilities. Autotrophic microalgaes can utilize carbon dioxide
as the carbon sources and sunlight as the energy for oil accumulation. Some
autotrophic microalgaes can be converted to heterotrophic microalgaes and such
heterotrophic microalgaes can accumulate biomass and oils using organic carbon
as its carbon source instead of carbon dioxide and sunlight. Moreover, algae can
be grown on marginal land using brackish or salt water. They do not compete for
resources with conventional agriculture and do not require herbicides or pesticides.
Besides, their cultivation could be coupled with the uptake of carbon dioxide from
industrial waste streams and remove the excess nutrients from wastewater. They
can be converted to biodiesel through enzymatic catalysis, acidic catalysis, or
alkali catalysis.
2.3 Production of Biodiesel from Microbial Oil

Microbial oils have been a potential alternative for biodiesel production due to
their similar fatty acid composition to the vegetables oils. In making biodiesel,
triglycerides in which three fatty acid molecules are esterifies with a molecule of
glycerol are reacted with methanol in a reaction known as transesterification or
alcoholysis. Transesterification is catalyzed by acids, alkalis and lipase enzymes.
As a result, methyl esters of fatty acids that are biodiesel are produced by
transesterification with glycerol as the by-product.
Table 2: Advantages and disadvantages of biodiesel oil that are produced from microbial oils.
ADVANTAGES DISADVANTAGES
able to meet the specifications established by high cost of production

the currently existing standards
exhibits some improved fuel properties to produce cells with a simultaneously high
 higher octane number growth rate and lipid content, they required
 lower viscosity due to the variability of  productive strains
microbial oil with a wide array and  optimized culture conditions
multiple combination in potential.
high growth rate of microalgae high cost and energy demand of harvesting
 satisfy the massive demand on biofuels unicellular algae
using land resources without causing  small cell size
any biomass deficit  dilute biomass produced (consumption
of water and energy and processing
costs)
cultivation consumes less water than land
crops
allows high-efficiency of carbon dioxide

mitigation due to the tolerance of microalgae
towards high carbon dioxide content in gas
streams
minimize the release of nitrous oxide which in

turn minimize the occurrence of acid rain
more cost-effective compared to conventional

farming
3.0 Methodology
3.1 Cheaper Production of Microbial Oil form Cheap Raw Materials
There is much advantage to produce microbial oil. The microorganisms breed
quickly, which can shorten the production cycle, the production is not affected by
season and climate and is easy to cultivate in large scale. However, impacted by the
source of raw material and the cost, the industrial production is at low level and the
economic feasibility is poor. Hence, looking for a cheap, high yield, clean and
environmental friendly microbial oil raw materials becomes the key to
industrialization of biodiesel. The cheap raw materials that can be determined are:
1) Carbon dioxide
Some microalgae can take up carbon dioxide as a natural carbon source for lipids
synthesis, which can subsequently be used as the feedstock for biodiesel
production. In an ideal sustainable microalgae cultivation system, the large
amount of CO2, water, and other nutrients are derived from renewable sources or
wastes from agriculture, municipality, or industry. The microalgae can be
harvested for lipid extraction, while the water can be recycled. In recent years,
extensive research, as summarized in Table 3, has been conducted.[9]
Table 3: Using CO2 as the carbon source for microbial oil production
Species CO2 Biomass Biomass Lipid cont. (%)
aeration (%, conc. (g L– productivity (g L–
V/V) 1) 1D–1)
Chlorella sp. 0.03 0.682 NA NA
Chlorella sp. 15 NA 0.53 34
Botryococcus 5 1.88 NA 36
braunii
Chlorella 5 1.55 NA 24.25
pyrenoidosa
Scenedesmus 10 NA 0.292 38.9
obliquus
Choricystis 5 NA 0.385 21.3
minor
Chlorella 15 NA 0.482 27
vulgaris
Nannochloropsis 15 2.23 NA 59.9
sp.
Note: NA = not available
2) C2 Compounds
Some small molecular organics like acetic acid, acetate, and ethanol, all of which
have only two carbons (C2 compounds), could be utilized by certain oleaginous
micro-organisms to accumulate lipids. The representative examples are
summarized in Table 4.[9]
Table 4: Microbial conversion of C2 Compounds into oils

Biomass
Carbon Lipid cont.
Species conc. (g L– Cultivation Time (D)
source (%)
1)
Crypthecodinium
Acetic acid 109 56.0 Fed-batch 16.7
cohnii
Cryptococcus
Acetic acid 2.85 25.8 Batch 4
albidus
Cryptococcus Sodium
2.79 24.5 Batch 4
albidus acetate
Cryptococcus
Ethanol 2.11 24.9 Batch 4
albidus
Candida curvata Ethanol 8.5 30.1 Batch 3.8
Chlorella Sodium
4.4 19 Batch 3.5
vulgaris acetate
3) Lignocellulosic biomass
As the most abundant organic sources in nature, lignocellulosic biomass has
attracted heightened research interest for microbial oils production. Some
representative examples are summarized in Table 5.[9]
Table 5: Microbial conversion of Lignocellulosic biomass into oils

Biomass
Species Carbon source Lipid cont. (%) Culture
conc.(g L–1)
Cryptococcus
Wheat straw 17.2 33.5 Batch
curvatus
Yarrowia Sugarcane
11.42 58.5 Batch
lipolytica bagasse
Trichosporon
Rice straw 28.6 40.1 Batch
fermentans
Mortierella
Rice hulls 5.6 64.3 Batch
isabellina
Trichosporon
Corn stover 15.44 23.5 Batch
cutaneum
Trichosporon
Corncobs 24.4 40.1 Batch
dermatis
Mortierella
Corn fi ber 18,2 45.7 Batch
isabellina
Populus
Rhodotorula
euramevicana 18.1 34.16 Batch
glutinis
cv leavesv
Aspergillus 36.6 mg/g dry Solid-state
Wheat straw NA
oryzae substrate fermentation
4) Industrial by product and organic waste

Converting industrial by-products and wastes containing plenty of organic carbons
into lipid is a sustainable option for recycling and conserving resources. Glycerol
is a good example. Glycerol can be utilized by most oleaginous micro-organisms
with similar lipid yield to that of saccharides. With the fast growing of biodiesel
industries, it will be largely generated as a by-product of the transesterification
process from the local plants, and thus is a promising cheap carbon source for
microbial oil production. Besides, other organic wastes, locally obtained from
agriculture, fishing, or manufacturing industries, can also be used. However,
compared with the biodiesel by-product glycerol, their complicated compositions,
especially some hard-to-degrade pollutants or toxics, continue to be a challenge.
3.2 Cheaper production of microbial oil form different technology of microalgae

production
One of the most sustainable ways to produce microbial oil is through cultivation of
microalgae. It has potential as biofuel feedstock because the lipid content and
biomass productivity of microalgae can be up to 80% and 7.3g/l/d based on the dried
weight of biomass respectively [14]. Microalgaes are well adapted in the
phototrophic culture condition.
3.2.1 Photoautotrophic production
Phototrophic algae has the advantage of using sunlight as free natural resources
under natural growth condition and assimilate carbon dioxide from the
surrounding air which is the cheapest carbon source for microbial oil
production. It is technically and economically feasible for large –scale of
microalgae biomass production which reduces production cost by increasing
the yield of microbial oil. Two system are being implanted based on the open
pond and closed photobioreactor technologies in large-scaled commercial
cultivation of microalgae .
3.2.2 Open pond production systems
Algae cultivation using this system can be categorized into natural waters or
artificial ponds. Raceway ponds are the artificial pond that is commonly used
nowadays. It usually lined with plastic or concrete which is about 20-35 cm
deep to ensure adequate exposure of sunlight to increase productivity of
microalgae [15]. Open pond system is the cheaper method for large-scale
microalgae biomass production because it required less capital equipments.
Open pond system can be implemented in marginal crop production area. Thus,
it does not need to compete with existing agricultural lands. They have lower
input energy requirement, easier regular maintenance and cleaning process
which return large net energy production. However, open pond system are less
efficient due to:
1) Evaporation losses
2) Temperature fluctuation in growth medium
3) Carbon dioxide deficiency
4) Poor mixing
5) Light limitation
6) Pollution and contamination
3.2.3 Close photobioreactor system
Microalgae production based on this system are used to overcome the pollution
and contamination problems associated with the open pond system.
Photobioreactor permits the species of microalgae for prolonged period with
lower risk of contamination. Therefore, its efficiency increases which in turns
produce high quality of microalgae and enhance the productivity of microbial
oil. Due to the higher cell mass productivity attained, harvesting costs can be
reduced. Closed photobioreactor system includes:
1) Tubular photobioreactor
2) Flat plate photobioreactor
3) Column photobioreactor.
Tubular Photobioreactor
Composed of airlift system and solar receiver [15]. Airlift system allow the
transfer of oxygen out from the photobioreactor and allow carbon dioxide
diffuse into the system [15]. Removal of oxygen will helps to reduce the photo-
oxidation and thus efficiency will be improved. Solar receiver is designed to
have high surface area to volume ratio to enhance the absorption of sunlight
which provide a platform to grow the algae [15]. The productivity of
microalgae will increases which returns to high yield of microbial oil. However,
they require large land space and have are some limitation of the length of the
tube which dependent on the oxygen accumulation, depletion of carbon dioxide
and variation in pH of the system along the tube. Therefore, they are difficult to
scale-up indefinitely.
3.2.4 Flat -Plate Photobioreactor
The system has high densities of photoautotrophic cells and large surface area
exposed to illumination to increase the exposure of sunlight [15]. The reactors
are made up of transparent materials for maximum amount of solar energy
being captured which enhance the photosynthetic rates [15]. Flat-plate
photobioreactors are suitable for outdoor mass culture of microalgae due to the
low oxygen build-up. The rate of productivity of microbial oil will increase.
Cheaper biodiesel production could be achieved.
3.2.5 Column Photobioreactor
Column photobioreactor system can produce cheaper biodiesel from microbes

(microalgae) because this system has the highest volumetric mass transfer of
carbon dioxide which results in increase yield of microalgae for microbial oil
production. The reduced photoinhibition and photooxidation helps to improve
dissolution and carbon dioxide fixation. Therefore, high efficiency is achieved.
Besides, this system is energy- saving because it only requires low energy
consumption, easy to sterilize and provides excellent mixing with low shear
stress. However, the construction of this photobioreactor is very sophisticated
and therefore it is costly.
4.0 Results and Discussions
4.1 Cheaper production of microbial oil form cheap raw materials
It has been well recognized that many oleaginous microorganisms have the
ability to convert multiple carbon sources into microbial oils. These carbon
sources, however, are different in the resources, characteristics, treatment
method, and fitness to oleaginous micro-organisms. Accordingly, diversified
routes can be expected in order to obtain microbial oils in a more effective way.
Table 6: Prospect of microbial oil form different sustainable carbon sources.

Carbon source Advantages Disadvantages, difficulties and
future efforts needed
Carbon dioxide Reduction of CO2 emission; Low efficiency in CO2 utilization
low cost and conversion into
microbial lipids; energy input for
CO2 supply; Smart bioreactor
design and microorganism
development are needed
C2 compounds Available from a variety of Further study is needed for
biodegradable organic comprehensive demonstration
wastes; effective conversion of their potential applications
was demonstrated in
some oleaginous micro-
organisms
Lignocellulosic Abundant and cheap organic Compositional complexity and
Biomass sources heterogeneity; Pretreatment
leads to inhibitive chemicals; Co-
utilization of
sugar components, and
comprehensive utilization of the
lignocellulosic biomass are
expected
Industrial by-products Abundant and available from Especially for organic wastes, the
and other organic industries; low cost compositional complexity,
wastes which is dependent on the
industrial process, may lead
to difficulties in microbial
degradation and lipid production
All sustainable carbon sources have its own advantages and disadvantages. However, the overall
economic feasibility needs to be evaluated further, with the advances in genetic engineering,
protein engineering, metabolic engineering, and process improvement, microbial oil from
various sustainable carbon sources will become one of the feedstocks for biodiesel production in
the future.
4.2 Cheaper production of microbial oil form different technology for microalgae
production
Table 7: Prospect of microbial oil form different technology of microalgae production

Production system Advantages Limitations
Open pond system
Raceway pond Relatively cheap Poor biomass productivity
Poor mixing , light and carbon
Low energy input dioxide
Utilization
Easy maintenance Limited to only few strain of algae
Easy to clean Cultures easily contaminated
Highly utilize non-agriculture land Required large area of land
Closed Photobioreactor system
Large illumination of surface area Some extent of wall growth
Good biomass productivities Fouling
Tubular Photobioreactor Suitable for outdoor cultures Required large area of land
Gradient of pH, dissolved O2 and
CO2 along the tube.
High biomass productivities Difficult to scale-up
Easy to sterilise Difficult temperature control
Small degree of hydrodynamic
Low oxygen build-up stress
Flat Plate Photobioreactor
Good light path Some extent of wall growth
Large illumination of surface area
Suitable for outdoor cultures
Readily tempered
Compact Small illumination area
Low cost Expensive compared to open pond
High mass transfer Complicated construction
Low energy consumption
Column photobioreactor
Easy to sterilize
Good mixing with low shear stress
Reduced photo-inhibition and
photo-oxidation
Closed photobioreactor could be attributed to more process control and potentially higher rate of
microalgae biomass production. Therefore potentially higher rate of biodiesel production and co-
products production. In conclusion, column photobioreactor system is the most efficient in
production of cheaper biodiesel from microbes (microalgae) because they are low cost, compact
and easily to operate.
5.0 Conclusion
Biodiesel, has increasing importance in human lives and its sources determine its usability.
Biodiesel, generated from renewable sources, is also environmental-friendly.
Microorganisms like algae (also microalgae), yeast, bacteria and fungi are chosen as
materials to produce vast quantities of high quality and purity biodiesel due to their lipid
accumulation ability and they are classified as “oleaginous” if the lipid contents exceed
certain percentage. The rapid growth rate of microorganisms, together with some other
characteristics and properties, makes them a good tool to produce very much microbial oils.
However, impacted by the source of raw material and the cost, the industrial production is at
low level and the economic feasibility is poor. Hence, looking for a cheap, high yield, clean
and environmental friendly microbial oil raw materials becomes the key to industrialization
of biodiesel. CO2, C2, Lignocellulosic Biomass, industrial by product and organic waste are
cheap sustainable carbon sources and have its own advantages and disadvantages. However,
the overall economic feasibility needs to be evaluated further, with the advances in genetic
engineering, protein engineering, metabolic engineering, and process improvement,
microbial oil from various sustainable carbon sources will become one of the feedstocks for
biodiesel production in the future.
Among the types of culture system for cultivation of microalgae, open pond systems have
dominated. However, they have low efficiency in microalgae mass production. Thus, several
types of closed culture systems such as tubular, flat-plate and column photobioreactors have
been developed which can show promise on the application in scaling up microalgae culture
for biofuel. Column photobioreactor possess many advantages for microalgae cultivation
such as high mass transfer rate, low energy consumption, good mixing with low shear
stress ,compact , no moving part and less land is required. However, problems arise when
scaling up this bioreactor. Penetration rate of light decreases exponentially as the distance
from the light source increases. Thus, an optimum column diameter must be determine to
compromise between productivities and capital costs in addition to considerations of space
required and the ease of cleaning. In future, new developments in lighting and scale-up are
required to make the closed culture systems more controllable, more cheap and more
EFFICIENT !!!
Reference
1. http://www.berkeleybiodiesel.org/advantages-and-disadvantages-of-biodiesel.html
2. http://www.esru.strath.ac.uk/EandE/Web_sites/02-03/biofuels/what_biodiesel.htm
3. http://www.epa.gov/air/caaac/mstrs/ciampa.pdf
4. http://www.biodiesel-energy-revolution.com/disadvantages-of-biodiesel.html
5. http://www.biodiesel-energy-revolution.com/disadvantages-of-biodiesel.html
6. http://answers.yahoo.com/question/index?qid=20130126083946AALGmt1
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Griffiths, Reay G Dicks, Christine Richardson and Susan TL Harrison, from Centre for
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Preparation’, Jingyang Xu, Wei Du, Xuebing Zhao, Guoling Zhang and Dehua Liu,
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Antonio A. Martins, Nidia S. Caetano, Science Direct (Online published)
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15. http://www.sciencedirect.com/science/article/pii/S1364032109002408

Table 1: Advantages and Disadvantages of Biodiesel Oil That Are Produced From Vegetables Oils

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TOWARDS CHEAPER MICROBIAL OIL FOR BIODIESEL PRODUCTION November 29, 2013

has lower toxicity drought happened during the dry climates

Biodegradable controversy concern about ‘we should plant

2.2 Microbial oil

2.3 Production of Biodiesel from Microbial Oil

able to meet the specifications established by high cost of production

allows high-efficiency of carbon dioxide

minimize the release of nitrous oxide which in

more cost-effective compared to conventional

Table 4: Microbial conversion of C2 Compounds into oils

Table 5: Microbial conversion of Lignocellulosic biomass into oils

4) Industrial by product and organic waste

3.2 Cheaper production of microbial oil form different technology of microalgae

3.2.1 Photoautotrophic production

3.2.2 Open pond production systems

3.2.3 Close photobioreactor system

3.2.4 Flat -Plate Photobioreactor

Column photobioreactor system can produce cheaper biodiesel from microbes

4.0 Results and Discussions

4.1 Cheaper production of microbial oil form cheap raw materials

Table 6: Prospect of microbial oil form different sustainable carbon sources.

Table 7: Prospect of microbial oil form different technology of microalgae production

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