Team:Slovenia HS/old
{{Slovenia_HS}}
Welcome to iGEM 2015!
We are the first high school
team from Slovenia
to compete at iGEM and are very excited to face the challenges
synthetic
biology poses. The team is composed of eight high school students
attending seven
different secondary educational institutions across Slovenia. Our
research is
performed at the Laboratory for Environmental Sciences and Engineering,
National Institute of Chemistry, Slovenia and at the Chair of
Biochemistry,
Faculty of Chemistry and Chemical Technology, University of Ljubljana
under
supervision of knowledgeable and encouraging mentors.
We have decided to tackle a
very imposing
ecological problem that is becoming more and more threatening due to
fossil fuel
depletion and a steady growth of energy consumption. Fossil fuels, such
as oil
and gasoline, have gained popularity in the beginning of the 20th
century,
following the industrial revolution. It is estimated that we have spent
between
100 and 135 billion tons of oil since 1850 and the demands are still
increasing. Fossil fuels are used in cars, airplanes and other
vehicles, to
power electricity plants, to heat our homes and to make many everyday
products,
such as medicines, cosmetics, plastics and synthetic fabrics. Our
society is
largely dependent on them, but fossil fuels take millions of years to
form and
are therefore non-renewable resources. According to some projections,
we only
have enough oil for the next 40 years, so it is becoming increasingly
necessary
to find an alternative method of obtaining fuels.
After a thorough discussion and
literature
survey, we have thus decided to make use of butanol. Research has
already shown
that because of its long chain and consequent nonpolarity, butanol can,
amongst
other uses, replace gasoline in internal combustion engines (it can be
used in
existing diesel engines without considerable modifications of the motor
system).
In nature, many organisms have
been proven to
be able to produce butanol (n-butanol or 1-butanol) by converting
glucose into
acids and then converting acids into alcohols. This organisms, mostly
bacteria
of the Clostridium genus (Clostridium acetobutylicum, Clostridium beijerinckii and Clostridium saccharoperbutylacetonicum),
however, have complex metabolism, slow conversion rate and are often
hard to
grow in laboratories or for industrious use. For these and other
reasons, they
are unsuitable for larger butanol production. On the other hand,
bacteria E.
coli have relatively simple and strikingly fast metabolism, already
researched
and utilized mechanisms for genetic manipulation and are relatively
easy to
grow and cultivate, making them the perfect laboratory and industrial
organisms.
We have sought help from the
Laboratory for Environmental
Sciences and Engineering at the National Institute of Chemistry in
Ljubljana,
as they have been engaged in advancing processes that enable conversion
of
waste and other renewable raw materials into energy (e.g. biogas to
syngas
transformation, bio-oil production through pyrolysis of waste etc.) for
a long
time now, and have recently also developed a process for the
biotechnological conversion
of biodegradable waste into hydrogen by means of anaerobic mixed
cultures grown
at special fermentation conditions. During this process a series of
intermediate products is produced, amongst others are high
concentrations of butanoic
acid. We had the idea of turning this organic acid into butanol, which
is
hydrophobic and can be separated from water medium by basic
concentration and
decanting processes.
That’s why we have
set on the path to modify
the E. coli in such a way, to
enable
transformation of butanoic acid to butanol. To aid in the process, we
will also
be using glycerol as a co-substrate, which can be readily obtained in a
suitable form as a side product of biodiesel production processes.
We will thus genetically
manipulate the E. coli bacteria
into performing only
the second phase of butanol production found in Clostridium bacteria
(acids to
alcohols conversion). By using biodegradable waste as substrate we
intend to
fully make use of all of the components involved in this processes
(anaerobic
microbial production of hydrogen), while subsequently (E.
coli fermentation process) also producing the much-needed
biofuel, thus enabling a simultaneous efficient and eco-friendly waste
management and energy production process. Our aim is to achieve the
highest
possible yields with our genetically modified in an optimized
bioreactor system.
In the first stage the biogas (H2)
will be isolated out of organic waste and suspended solids will be
produced
which may be used as a natural fertilizer. At the same time the
butanoic acid
formed will be redirected in another bioreactor (containing our
modified E. coli) where it will be
converted into
bio-butanol.
Up to now, we have cloned three
genes coding
for CTFA, CTFB and BDHB proteins (in PSB1C3 vector), that are
responsible for
butyrate aldehyde to butanol in Clostridium
acetobutylicum. We further intend to make composite
constructs to include
this genes in vectors with promotors and ribosomal binding sites to
allow for
expression of these genes in E. coli.
In the biotechnological
laboratory we have
performed an initial screening of growth conditions, such as butanoic
acid
concentration and butanoic acid / glycerol molar ratio, as well as
comparing
the growth in the presence and absence of oxygen. We are working with
two
negative controls, (i) E. coli
strain
Dh5 alpha and E. coli strain Dh5 alpha with PSB1C3 plasmid containing
only the
promotor and the ribosomal binding sites (i.e. with no genes included),
to
provide proof E. coli does not inherently convert butyric acid into
butanol. We
intend to perform similar tests with our final microorganism
– modified E. coli
and provide proof of concept for the said fermentation in a bench-scale
laboratory bioreactor.
We are also looking for other
teams with
similar projects for possible collaborations, so if you are interested,
we
encourage you to contact us!
Before you start:
Please read the following pages:
Styling your wiki
You may style this page as you like or you can simply leave the style as it is. You can easily keep the styling and edit the content of these default wiki pages with your project information and completely fulfill the requirement to document your project.
While you may not win Best Wiki with this styling, your team is still eligible for all other awards. This default wiki meets the requirements, it improves navigability and ease of use for visitors, and you should not feel it is necessary to style beyond what has been provided.
Editing your wiki
On this page you can document your project, introduce your team members, document your progress and share your iGEM experience with the rest of the world!
See tips on how to edit your wiki on the Template Documentation page.
Templates
This year we have created templates for teams to use freely. More information on how to use and edit the templates can be found on the Template Documentation page.
Tips
This wiki will be your team’s first interaction with the rest of the world, so here are a few tips to help you get started:
- State your accomplishments! Tell people what you have achieved from the start.
- Be clear about what you are doing and how you plan to do this.
- You have a global audience! Consider the different backgrounds that your users come from.
- Make sure information is easy to find; nothing should be more than 3 clicks away.
- Avoid using very small fonts and low contrast colors; information should be easy to read.
- Start documenting your project as early as possible; don’t leave anything to the last minute before the Wiki Freeze. For a complete list of deadlines visit the iGEM 2015 calendar
- Have lots of fun!
Inspiration
You can also view other team wikis for inspiration! Here are some examples:
- 2014 SDU Denmark
- 2014 Aalto-Helsinki
- 2014 LMU-Munich
- 2014 Michigan
- 2014 ITESM-Guadalajara
- 2014 SCU-China
Uploading pictures and files
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server. Remember to keep all your pictures and files within your team's
namespace or at least include your team's name in the file name.
When you upload, set the "Destination Filename" to Team:YourOfficialTeamName/NameOfFile.jpg
.
(If you don't do this, someone else might upload a different file with
the same "Destination Filename", and your file would be erased!)