Table of Contents

Preface Page 3
Introduction Page 4
Process Page 5
Algae cultivation Page 7
Mozambique Page 8
Budget biodiesel project Page 10
Plan Page 11

2
 Preface
An unique opportunity, motivated people and a huge challenge: The Imagine project. In March of 2004 our
biology teacher told us about an interesting project that was just launched. Beside the fact that you could use the
project for your “profielwerkstuk” (a piece all exam students in secondary schools have to make) there were also
captivating and inventive subjects which you could choose from. Furthermore there was excellent assistance and
experienced guides and last but not least, we could win a trip to a developing country.

Let’s introduce ourselves first. We are Sonja Boas and Chang Liu. We are classmates and we are going to the
final year of high school. We both live in Soest and we cycle every day together to The Baarnsch Lyceum. In
Dutch high schools there is a system where students can choose the direction of education which in they are
interested. We are both following the natural sciences direction and are interested in all that lives. It should not
be a surprise that we both want to study medicine.

During the International Biotechnology Congress in Ede the five promising propositions were presented to us.
After many discussions we decided to continue with the biodiesel proposal. In the course of the project we
learned more and more about the subject. We got to know our guides and the scientists who proposed the
biodieselplan better in a short time as well. We want to seize this occasion to thank Daan Schuurbiers, Marije
Blomjous and Tycho Malmberg for all their assistance and patience in answering all our questions. Also we want
to give thanks to Wouter van Winden and Bram van Beek for their wise advices, great tips and professional
knowledge.

3
Introduction
Our world is dying, and we are the cause of it. We pollute her air and we poison her water. We exploit her and
take from her all that she has build up in the thousands of years and use it up in just a few decades. Millions of
factories and cars emit thick smoke day and night. Not only does this pollute the air but the soil too. It even
damages the ozone layer and provokes the notorious green house effect. We have to realise that if we go on this
way, we will devastate Mother Earth and therefore ourselves. For that reason changes in our lifestyle must be
made.

We think that an important problem for our environment lies with the production and use of fuels. First of all we
aim at diesel but in the future other types of fuel could be converted into ecological friendlier products. For the
production of contemporary fuels fossil reserves are used. Eventually, this natural source will not be sufficient
anymore for our gigantic consumption. Additionally the production process creates an enormous emission of
carbon dioxide. When the diesel is burnt in the engines, more carbon dioxide will be released into our
atmosphere. The concentration gets too high and the greenhouse effect is worsened.

Thanks to the Imagine project we encountered a good solution. Wouter van Winden and Bram van Beek
submitted a proposition, which we think that if it could be realised, it would be a large step to a durable lifestyle.
They wanted to make fuels with algae as primary product. This cleaner fuel is called biodiesel. Biodiesel is
already used in several countries. In Germany for example cars drive on a mixture of 60% of conventional diesel
and 40% biodiesel. The experiences so far are highly positive. As yet biodiesel is too coarse for most of the
diesel engines to run on exclusively. But even if 40% of all diesel would be replaced by biodiesel, a great part of
our goal would be achieved.

The biodiesel in Germany is made from rapeseed. However it is impractical and not lucrative to produce
biodiesel from rapeseed on a large scale because the cultivation of it takes in too much space. That is why we
want to use algae as material for our biodiesel. Algae produce relatively much more oil than rapeseed. Some
types of algae can convert up to 70% of their total biomass into oil. Algae for our fuels will make us less
dependant of the fossil fuels. Also the production process of diesel from algae is much better for the nature. The
algae reuse all carbon dioxide that is emitted during the process for more oil production. The exhausts of
biodiesel are cleaner than that of “fossil diesel” as well. In a word we can say that biodiesel is a promising
candidate for sparing the fossil fuels and reducing the current immense pollution.

In this report we will elaborately discuss a few concerns. First we will show how the process works and that it
can be realised. The cultivation of the algae is an important part of this process; it will therefore get the needed
attention. Because our goal is to make this project successful, we made an extensive project plan. In this plan we
thoroughly examined how the project can be achieved effectively. To support this, we tried to work out the
financial aspects as much as we could and adapted it to a realistic location. This location is selected carefully
according to several criteria. These criteria will be discussed in this report too.

We believe in the plan but we are the pioneers of this method. We will make the first step and we will surely
encounter many problems on our path. Especially the financial matters will be a challenge. However, where
there is a will, there is a way and we will follow it. We are confident that in the future solutions will be found for
all problems. We see a future for biodiesel.

4
Process
Every day we want to produce 10 litres of biodiesel. The preparation of biodiesel from algae is a complicated
process. It contains many steps. We subdivide this process in four parts. We will clarify these four steps as
comprehensible as possible. In figure 1 of the appendix is a design of this process enclosed.

A. Photosynthesis of sugar canes

B. Alcohol from sugar canes
C. Oil from algae
D. Biodiesel from oil and alcohol

A. Photosynthesis of sugar cane

Sugar canes contain, just like all other plants, chlorophyll granules. In these granules photosynthesis takes place.
This is a well-known process and already thoroughly studied. During this process the plant takes up carbon
dioxide from the air and converts this in its granules into other products with the aid of sunlight. The two final
products are oxygen and glucose. Below we will represent this process with the reaction formula

B. Alcohol from sugar canes

From the last process we need the end product glucose. In sugar canes is a lot of glucose stored and that is why it
is a suitable plant for our production process. From this glucose we will make alcohol. This will happen by
means of yeast. Yeast is a micro-organism that belongs to the single celled fungus. The scientifical name for
yeast is Saccharomyces cerevisiae. Yeast decomposes glucose into carbon dioxide and alcohol. This process is
called fermenting. The less oxygen there is involved in this process, how better the circumstances for the
efficiency of the yeast. Below we will represent this process with the reaction formula

The carbon dioxide that is released here will be used later in the process for the cultivation of the algae. The
obtained alcohol will be further processed in part D.

C. Oil from algae

We have to acquire the oil out of the bred algae. There
are only a few experiments available to serve as our
model. In such a way the oil from rapeseed is obtained
by a combination of extraction and filtration. First the
rapeseed is heated to increase the fluidity of the
captured oil. Now the oil is extracted from the rapeseed
tissue. Subsequently the oil is filtrated to remove any
solid fragments. We assume that we can use a similar
method to win oil from algae. First a part of the algae
have to be filtrated out of the breeding pond. This part
will amount to 50% of all the algae in the pond every
twenty-four hours because the algae duplicate every
day. Every month the concentration algae in the pond
will be checked. After filtration the oil will be extracted
from the algae just like with rapeseed. The oil will be Rapeseed cultivation

5
distilled to remove the remaining of the algae. Distillation is a better method than filtration in this case because
we can withdraw the last bits of oil out of the algae and remove solid bits. This will raise the output. The two
final products are now oil (triglyceride) and algae pie. This pie can be used as cattle feed or fish feed. The algae
in the breeding pond can also reuse it.

D. Biodiesel from oil and alcohol

Now it is time for the last step. In part B we obtained alcohol and in part C oil. Now we are going to put those
two products together in a reactor to produce usable fuel. This oil was namely not good enough yet because the
viscosity was too high due to the glyceride-connections. That is why we have to remove the glycerol from the
triglyceride. We do this with the aid of alcohol. The OH-group of alcohol will bind with the detached, aloof C-
atom of the triglyceride. Moreover the two remaining carbon atoms of the alcohol will bind to the remaining
ester. The result is two end products: glycerol and three fatty acids. The fatty acids constitute our biodiesel. This
process is presented below with the reaction formula.

Now we have a mixture of glycerol and biodiesel. The last step is the separation of these two products. This will
be done with distillation.

6
Algae cultivation
The algae are the indispensable link in our project. We have to win the main
constituent out of the algae, namely oil. Algae absorb carbon dioxide and
nutrients from the water and make oil out of their biomass, while they produce
oxygen. Algae reproduce themselves nonsexually; they divide themselves into
two cells, grow bigger and divide again. We have chosen a type of algae that
accumulates a lot of oil and satisfies a few requirements. Some of these
requirements involve their habitat. These criteria will be worked out when we
discuss the location of the project.

The alga that we preferred was the Botryococcus braunii BBG-1. On the left
there is a picture of this specie. The Botryococcus braunii have green cells with dimensions of 15 to 20
micrometers and an oval shape. They form star shaped colonies. This specie has one of the highest oil
productions. Under optimal circumstances the Botryococcus braunii can achieve a share of 61% oil out of their
dry body mass. Under stress this will reduce to 31%. Relatively this is a excellent performance and since the oil
production is essential for the success of our project it makes the Botryococcus braunii our number one alga. The
braunii has an maximum growth when the temperature is between the 22 and 25 . Just like all other algae the
Botryococcus braunii needs minerals as nitrate and phosphate. In the appendix in figure 3 there is a list of
important nutrients that optimize the habitat of the algae. Natural seawater contains all of these nutrients and do
not have to be added. It also appears that the Botryococcus braunii can survive under poor circumstances.

The next item we have to consider is that the algae have to be bred on salt water. For this water we use seawater.
First of all natural seawater contains as said nearly all of the needed nutrients. Second of all it is affluently
available. We are going to breed the algae in a circuit-shaped pond. In the appendix in figure 2 it is represented
schematically. The width of the pond will be around the 23 meters and 76 meters in length. The height will be
around 15 cm. The total space the pond will take in will be 1750 square meters. Detailed construction
information can be found in the appendix (figure 6). The algae must remain compounded and therefore the water
must be constantly stirred. To reach this we will install a pump that keeps the water circulating. There is
insufficient nitrate and phosphate in the seawater and we have to insert more of those minerals additionally. For
this we can use the wastewater from the factories in the neighbourhood. Beside the fact that there is enough
nitrate and phosphate in this water for the algae, we can also keep the salt concentration on a certain level.
Further we can heat or chill the water with a regulation system before we add it to the pond to control the water
temperature. We will connect a thermometer to the system to watch the water temperature.

Like oxygen is essential for human beings, carbon dioxide is for algae. We must add it to the algae’s habitat.
Like said before there is a alcohol factory nearby the cultivation where alcohol is produced from sugar canes.
The only waste product is carbon dioxide. We will seize this and conduct it into a pipe through a compressor.
This compressor will give the gas a higher pressure and a smaller volume. The tube leads to a sparger that is
zigzag installed into the bottom of the pond where the carbon dioxide gas will bubble through the water to the
surface. On account of the zigzagging structure of the sparger all flowing algae are provided with carbon dioxide.
The CO2-bubbles are multifunctional in this system. First of all the algae absorb the carbon dioxide for their
growth. Second of all they can loose their oxygen to these bubbles which prevents the algae to get stressed.
Finally through this gas-exchange a perfect buffer arises. The oxygen binds hydrogen atoms that can make the
water too acid. H2O is the product of this reaction and the pH stays on a constant level. Below you can find the
reaction formula.

We can watch the pH with a pH-meter. This will be connected to the sparger which adds the CO2. In this way
the concentration of the buffer can be controlled.

7
Mozambique
An important part of the project is of course finding a good location. We looked carefully at the process and
examined which geographical factors will be important. The first demand was that it must be a developing
country. Now we could exclude many countries. The next demand is that there must be enough sugar cane
cultivation. If there is sugar cane, there is alcohol production and since alcohol is one of the main components of
the process this is a necessity. Now just a few countries are leftover. The finalists were Cuba Brasil and
Mozambique. But there are more criteria. Cuba was excluded because of political instability. The next factor was
the presence of natural seawater. We need this water for the algae breeding pond. Brasil and Mozambique
suffices both but we decided to go on with Mozambique because our motivation to help this country is bigger.

Now we had to choose a specific place in Mozambique. Without doubt this place must lie to the coast. Second of
all it has to be near sugar cane plantages. The last but surely very important factor is the temperature. Our algae
feel the most comfortably when the temperature is between the 22 up to 25 . The temperature in the whole
year must be as close as possible to this level. After some research we found that the middle part of the coast line
came closest to these temperatures through the whole year. Beira is at the coast in the middle of Mozambique
and had the most favorable temperatures. Beside that Beira is also the second great port of Mozambique and the
infrastructure is relatively developed. In the appendix (figure 4) is a monthly statement of temperature and
precipitation in the city of Beira. Not much to the north there are sugar cane plantages. We have found our
destination.

History
Colonisation
Mozambique lies in the south-west of Africa. The Portugeze
explorer Vasco da Gama discovered Mozambique on his journey
to India in 1498. In 1505 The first military posts arrived at Sofala
(province where Beira lays today). Mozambique became a
Portugeze colony. The colonists were interested in the gold and
ivory from the midlands of Mozambique. They first lives of the
slavery and later they exported agricultural products and lived of
the transit of products from Rhodesia (the current Zimbabwe) and
South-Africa. The country was fully exploited during this
colonization. There was little to no attention paid to the local
development of the infrastructure for example, or the proficiency
of the natives.

Decolonization
The decolonization went violently because the Mozambiquans
lived very poor under the colonial system and they saw how the
Portugeze took their riches and posessions. They saw that the most
fertile land was owed by the white farmers and they had to use the
dry, infertile pieces of ground to stay alive. Inevitably protest
groups were formed. The Frelimo (Frente de Libertacao de
Mocambique = Front for the liberation of Mozambique) was a
succesfull protestgroup. It was founded on the 25th of June in 1962
by Eduardo Mondlane (among others). A ten-year battle for
independance arose between the Mozambiquans and the colonists.
Eventually in 1975 liberty was reached. Mozambique was
independant.

The joy however was only of short duration. The Portugeze left the
country suddenly in 1975 without preparing the country for it. All
educated and skilled men were taken away. To give you an idea: In
the whole country there were ten teachers left. The Frelimo
became the nieuw Mozambiquean government and they embraced
the socialism. With this they closed tight ties with the Soviet Union.

8
Unfortunately, socialism failed in Mozambique and a group named the Renamo protested. They destroyed
schools, public buildings and the little infrastructure that Mozambique posessed. Finally the Peace Treaty of
Rome was concluded in 1992. At that point in time Mozambique was a havoc. Now it had to be rebuilt. The
socialistic system was replaced by a free market economy. In 1994 the first democratic elections were held since
years.

Future
Even though Mozambique achieved a lot in the passed few years and it now may call itself a independant
democracy, it remains one of the poorest countries in the world. But this is not surprising. First of all the country
has been free only just since 1992. When it was time to make a new start they had to begin with nothing at all
because everything was demolished. There was barely education because there were no educators. Even after
their libertation, fortune does not seem to be on their side. Drought, floods and famine afflict the nation. As a
result of the floods in 2000 the international society gave Mozambique US$ 450 million for rehabilitation and
unburdened them of debt.The Netherlands donated besides calamity aid with a cash value of US$10 million, an
amount of money of US$45 million to support the reconstruction. Mozambique put this money into Education
and Water and Sanitary from their reconstruction programm. The floods in 2001 in Central Mozambique led to a
second reconstruction programm of US$132 million. The Netherlands contributed US$13 million.

We can say that Mozambique receives much financial support in the reconstruction of their country. The
economy grows as one of the fastest in the world (also because of the low starting point). This gives us hope and
we can also contribute to a better future for Mozambique. If biodiesel production will be a important point of
attention in the world, then Mozambique will be the pioneer of it.

9
Bugdet Project Biodiesel
Entry Specification Total costs in euro's

Land rent 5 000

Algae pond 10 000
Pond
Pump
Algae
Thermometer
pH-meter
Seawater transport
Nutrients 2 000
Heating installation
Chilling installation
Carbon dioxide 7 000
Compressor
Sparger
Oil production from algae 15 000
Filter
Extractor
Distillator
Alcohol 2 000
Biodiesel production 5 000
Reactor
Distillator
Automation 10 000
Laptop
Telephone-/internetconnection
Energy (electricity) 3 000
Distribution 6 000
Storage reservoir
Transportation
Employee 20 000
Salary
Adventitious 25 000

Total budget 110 000

For all transport we engage another company. For the budget we must find a sponsors. We can approach
companies which support developing countries or we can adress ourselves to nature organisations like
Greenpeace. We also can appeal to the EOF; the European Development Fund since this organisation offers
financial support to various projects in developing countries. Finally we can also conclude a cheap loan at EIB;
the European investment bank. This organization also offers cheap loans for projects in Third World countries.

Once the production is on pace we will get incomes from it. First of all we will be able to sell the biodiesel.
Secondly we can sell the remained alga wafer as cattle fodder or fish fodder.

10
Plan
To give the project a maximum chance, we must work out all of the details as well as possible. Here we will
make a good start by discussing the most important matters step by step. Of course during the implementation it
will become clear where the difficulties lie and what appears to run prosperous. The pilot system will be well
observed for a year. Afterwards we will look at the results and decide together what potential possibilities the
future of biodiesel holds. Now we will start at the very beginning and find a suitable destination for our project.

Location
As already presented in the report, Mozambique is a promising location. Not only it is a developing country by
the sea, but it also has suitable temperatures. Moreover, nowadays there is a political stable environment in
Mozambique and the economy grows tremendously. We will now discuss the location more specific. The area
between the cities Beira and Quelimane proved to have the best circumstances for this project. In that area we
must find a piece of land. Since there are many farmers who live there and cultivate their ground for own
consumption, this must be no problem. If we would rent a piece of soil of approximately a hectare of a farmer,
this would bring advantages for both sides. First he would receive rent from us, what will be a relatively large
amount of money for him (the average year income amounts to $250, - we apprehend a hiring budget of $5000,-).
Secondly we can adopt the farmer himself as an employee of the project and he would receive a high salary.

The presence of an alcohol factory is also of importance. Since there are many sugar plantations in the area it is
assumable that there is an alcohol factory. We have to approach them and make a deal with them. First of all we
must be allowed to capture the carbon dioxide they produce and use it for the algae. Probably this will not be a
problem since this gas is nothing but a waste product for them. Second of all we would be a consumer of their
alcohol for the last step in our biodiesel production process. Annually we will be needing approximately 3000
liters of alcohol, depending on the performance of the algae to produce oil. Beira is a large city and it is no
wonder that there are many factories around it, for example a steel factory. We will need the waste water of one
or more of these factories. It contains namely nitrate and fosfate that our algae need for their growth. We
presume that they will allow us to use their waste water. Possibly they can even transport it for us for a small
compensation.

Algae
What are vital for the process are of course the algae. There are many kinds of algae which all require different
life environments. We choose the Botryococcus braunii for our project. Our pioneer algae must be healthy and in
top condition. That is why we will order them at a specialised organization. CCAP is an acknowledged firm for
these matters. CCAP stands for Culture Collection of Algae and Protozoa. It is the longest established of the
world's major protistan service culture collections. It holds over 2000 strains of algae and protozoa which are
supplied worldwide to industry and the academic community. They can deliver us the Botryococcus braunii. For
more information you can go to their website: http://www.ife.ac.uk/ccap/index.html

Material
Now we must think about the materials. Our first concern is the algae breeding pond. The material must be solid,
durable, stainless and impermeable. A good candidate is stainless steel and since there is a metalfactory in the
neighborhood we can make the pond there. The tube installations can be made of a metal. The high-tech
materials like the hydraulic pump, filter, extractor, distillators, compressor, chilling-/heatingsystems and a
reactor we must find in a more developed area. South-Africa would be a good source
for these instruments. The hydraulic pump must be able to pump the water around 2
meters per seconde. Since the length of the pond “circuit” is approximately 131
meters the pump must move 150 000 liters of water per 65.7 seconds. There are
pumps that meet this requirement. Techinally qualified pumps can be found at
http://wholesalehydraulics.com/AFPProducts/Products.asp?c=5 A suitable pump
that can be found on this site is the DPG2, produced by different firms.

Construction
After all the materials are complete we have to start the construction. We will ask an expert from The
Netherlands to go with us and lead the construction. The construction workers we can find in Mozambique.
Africon will be a good source of professional experienced men. They have worked several times before with
Mozambique so they are familiar with the people and culture. It would be a great integration aid for us when
Africon would be involved in the construction. More information about Africon can be found at their website:
http://www.africon.com/T We start with the breeding pond. The pond must be installed firmly into the ground in

11
a sunny place. Mathematical information about the pond can be found in the appendix. It must be as much as
possible isolated from other organisms. The carbon dioxide installation must be connected to the sparger, which
is installed in the pond. The other side must be connected to the compressor, where the CO2 is stored. The
hydraulic pump must be placed. The addition of nutrients will be done with the waste water. Each day we need
to refresh 5000 litres of waste water to suffice in the algae’s needs. This water will first pass the cooling-/heating
system so the water temperature in the pond can be controlled.

To start the process the algae must be taken out of the pond. This will happen through a filter. With the filter we
take half of the amount of algae out of the pond every day. The Botryococcus braunii reproduces at least one
time per 24 hours provided that it has enough nutrients and sunlight. So the concentration algae on the moment
you start filtering must be constant every day. The following step is to install the extractor and the distillator. The
oil that is finally made pure by the distillator will go through a direct connection to the reactor. Next to the oil
supply is a connection made for the alcohol supply. Finally the reactor is connected to a distillator. This will
finish the process and produce ready-made biodiesel. In the appendix (figure 5) you can find a schematic plan of
the system.

Distribution
Since the present-day diesel engines can not yet drive on pure biodiesel, but only on mixed diesel (conventional
diesel and biodiesel it is a advantage tot be in Beira. Here is namely a large oil connection to the inlands where
we may be able to include our biodiesel into the existing diesel in the future. In this way we can sell our product
directly and it will be surely consumed .If this possibility is not available for some reason; we will have to find
other ways to turn over our biodiesel. We hope that it
will be mainly for the inlands. We will begin with
conquering the market in the bigger cities, expecting
the smaller towns and inland countries will follow.
For the transport we will approach a transport
company.

Obstacle
An obstacle where we have less hold on than the
other matters is the financial support. Nonetheless we
have good hope because many countries in the world
put a lot of investments in projects in developing
countries. The UN-countries are even obliged to
donate an amount of money for these projects or countries annually. We hope that the people realise the
importance of this project for the future of our environment and are willing to offer their support in any way.
Thus we can appeal to companies or organizations that focus on a better environment. Another option is the EOF,
the European Development Fund. This fund can offer financial support for projects in Third World countries. If
needed we can obtain a cheap loan at the EIB, the European Investment bank, which offers particular interests
for projects in developing countries. There are therefore many possibilities.

Pioneers of the 21st century

It is proven that the process works. Theoretically this pilot works without problems and many litres of biodiesel
will see the light every day. That is why we have to construct this system precisely so we can prove to the rest of
the world that it is also possible. Many steps of the process are already studied and proven. A few examples are
producing alcohol from sugar canes, oil from algae, breeding algae and the reaction between oil from algae and
alcohol. What we are doing is fixing all these steps into a complete system. We hope we get the chance to show
this to the rest of the world so they can see it with their own eyes.

It is improbable that biodiesel will be produced on a large scale before the fossil fuels are exhausted. The
production of biodiesel is namely more valuable than the production of fossil fuels. Yet we must be prepared
because it will not be long before the natural resources are worn out definitively. That is why we have to develop
the technology now and master it so when the moment comes we will be prepared to switch over to the eternal
source of biodiesel.

Now that Mozambique has reached the position of a free, independent state, it can begin with the reconstruction
of the country. The nation can contribute to this by continue doing what they were successful in a broader
context, namely fighting for freedom; this time the freedom of Mother Earth and mankind.

12
13
Figure 1
The process

14
Figure 2
The breeding pond

15
Figure 3
Table of nutrients for algae

1000 ml of seawater
NaNO3 2 mM
NaH2PO4 0.1mM
ZnCl2 1 µM
MnCl2 1 µM
Na2MoO4 1 µM
CoCl3 0.1 µM
CuSO4 0.1 µM
Ferric citrate 20 µM
Na2-EDTA 26.4 µM
Thiamine* 35 µg/l
E.
Biotine* 5 µg/l F.
pH: 7.5-7.7 G.
H.
I.
J. Figure 4
Table of temperatures of Beira
BEIRA 19 80 S, 34 90 E, 52 feet (16 meters) above sea level.

Jan Feb Mar Apr May Jun Jul Aug Sep Oct Nov Dec

Avg. Temperature 28 27 27 25 23 21 20 22 23 25 27 27

Avg. Max Temperature 30 30 30 28 27 26 25 26 27 28 30 30

Avg. Min Temperature 24 24 23 22 19 17 16 17 19 21 23 24

Avg. Rain Days 5 5 7 4 2 1 2 2 2 2 3 4

Avg. Snow Days 0 0 0 0 0 0 0 0 0 0 0 0

Figure 5
The complete process

16
17
Figure 6
K. The dimensions of the breeding pond

Now we are going to lay out the calculations which led to these dimensions. First of all the
surface of the pond must be 1000 square meters. We choose 30 meters for L and 10 meters for
B.

The two round ends of the pond must cover a surface of 1000 – 600 = 400 square meters. You
can calculate the radius with the formula for the surface of the two ends of the pond:

18