Latest revision as of 18:14, 28 October 2013

PHAECTORY: Good reasons for a new chassis

The widely spread diatom Phaeodactylum tricornutum is particularly interesting. Its entire genome has been sequenced, an easy biolistic method for transfection is well established as well as protocols for the cultivation are available. Taking this into account it appeared to us as the perfect organism to produce complex proteins for the iGEM competition.

"Classical" large-scale biotechnology is mostly applied in bacteria and other fungi. Both need to be fed with carbon sources and the purification of the desired products e.g. functional high-value proteins is cost intensive, very time consuming and difficult. This is due to the fact that the cells need to be disrupted hence proteins/metabolites have to be removed from the medium.

By using a green cell factory of plants or algae it is possible to produce all kinds of products even high-value proteins like human antibodies in a large-scale background. Looking at previous iGEM-projects we recognized that no practical green chassis was introduced into the competition although these organisms perform photosynthesis, which enables a carbon dioxide neutral handling. In the photosynthetic process light energy is converted into chemical energy, which can be used to fuel different cellular reactions in the cell. Concerning the problem of global warming these organisms offer the possibility to reduce the emission of green house gases and work against the extensive use of fossil fuels. Moreover, it is not necessary to feed green algal cell factories with carbohydrates. The world population is continuously growing resulting in a scarceness of nutrition. The iGEM team Marburg aims at introducing a green cell factory, which is driven by sunlight, produces no carbon dioxide and above all does not compete with the nutrition-producing sector.

The microalga P. tricornutum is a green organism incorporating all above-mentioned advantages. This fast-growing organism lives with carbon dioxide and light and is able to produce substances like bioplastic, biofuels, spider silk and other complex proteins (Hempel and Maier, 2012, BIOforum). This year, the iGEM team Marburg decided to produce a human antibody against the Hepatitis B virus (PHAECTORY). An important feature of PHAECTORY is that the antibody is directly secreted into the medium, which enormously simplifies purification. In general, microalgae secret only very few additional substances and therefore all products produced in PHAECTORY are already present in a high purity (Hempel et al., 2011, Plos One).

In order to introduce this organism as a chassis to the iGEM community, we build a toolbox, which contains expression vectors, promoters, selection markers, reporter proteins and signal peptides for P. tricornutum. This allows every future iGEM team to produce their own complex proteins, which can easily be processed.

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-'''Phaectory'''
+PHAECTORY: Good reasons for a new chassis <html><a href="https://2013.igem.org/Team:Marburg/Project:Challenge"><img src="https://static.igem.org/mediawiki/2013/7/71/Mr-igem-next-arrow.png" style="float:right;"></a></html>
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+{{:Team:Marburg/Template:ContentStartNav}}The widely spread diatom ''[[Team:Marburg/Project:Ptricornutum|Phaeodactylum tricornutum]]'' is particularly interesting. Its entire genome has been sequenced, an easy biolistic method for transfection is well established as well as protocols for the cultivation are available. Taking this into account it appeared to us as the perfect organism to produce complex proteins for the <html><a href="https://igem.org/" target="_blank">iGEM competition</a></html>.
-'''Phaectory: Antibodies grown with sunlight'''
+"Classical" large-scale biotechnology is mostly applied in bacteria and other fungi. Both need to be fed with carbon sources and the purification of the desired products e.g. functional high-value proteins is cost intensive, very time consuming and difficult. This is due to the fact that the cells need to be disrupted hence proteins/metabolites have to be removed from the medium.
-The use of proteins in medical treatment and diagnosis is steadily increasing. Many pharmaceutically relevant proteins, such as antibodies, have a complex biological structure with specific posttranslational modifications. Furthermore, these proteins need to be highly pure because of their use in diagnostics and treatment. Therefore, a major challenge is the development of systems that produce pure, posttranslational modified proteins - best at low costs.
+<html><center><a href="https://2013.igem.org/Team:Marburg/Project:Milestones" title="PHAECTORY"><img src="https://static.igem.org/mediawiki/2013/1/1c/Phaectory.png" alt="PHAECTORY" /></a></center></html>
-Recently, the sunlight-driven microalgae Phaeodactylum tricornutum has been established as an elegant tool for the production of proteins. A major - however yet unchallenged - advantage of this system is the production of advanced proteins with specific posttranslational modifications and the ability of their direct secretion into the medium allowing easy purification with low effort and costs. Our mission is to challenge Phaectory for the convenient production of antibodies and make the system accessible to following iGEM teams.
+By using a green cell factory of plants or algae it is possible to produce all kinds of products even high-value proteins like human antibodies in a large-scale background. Looking at previous iGEM-projects we recognized that no practical green chassis was introduced into the competition although these organisms perform photosynthesis, which enables a carbon dioxide neutral handling. In the photosynthetic process light energy is converted into chemical energy, which can be used to fuel different cellular reactions in the cell. Concerning the problem of global warming these organisms offer the possibility to reduce the emission of green house gases and work against the extensive use of fossil fuels. Moreover, it is not necessary to feed green algal cell factories with carbohydrates. The world population is continuously growing resulting in a scarceness of nutrition. The iGEM team Marburg aims at introducing a green cell factory, which is driven by sunlight, produces no carbon dioxide and above all does not compete with the nutrition-producing sector.
-[[File:Fabrik horizontal.png|1000px|center]]
+The microalga ''P. tricornutum'' is a green organism incorporating all above-mentioned advantages. This fast-growing organism lives with carbon dioxide and light and is able to produce substances like bioplastic, biofuels, spider silk and other complex proteins (Hempel and Maier, 2012, BIOforum). This year, the iGEM team Marburg decided to produce a human antibody against the Hepatitis B virus (PHAECTORY). An important feature of PHAECTORY is that the antibody is directly secreted into the medium, which enormously simplifies purification. In general, microalgae secret only very few additional substances and therefore all products produced in PHAECTORY are already present in a high purity (Hempel ''et al.'', 2011, Plos One).
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+In order to introduce this organism as a chassis to the iGEM community, we build a toolbox, which contains expression vectors, promoters, selection markers, reporter proteins and signal peptides for ''P. tricornutum''. This allows every future iGEM team to produce their own complex proteins, which can easily be processed.
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-<li class="p1"><div><a class="a1" href="#p1"><b>INSECTS</b></a><img src="http://www.cssplay.co.uk/menus/horizontalaccordian/pic1.jpg" alt="" title=""><span>INSECTS : Some descriptive text can go here including <a href="index.html">links</a>.</span></div></li>
-<li class="p2"><div><a class="a2" href="#p2"><b>MAMMALS</b></a><img src="http://www.cssplay.co.uk/menus/horizontalaccordian/pic2.jpg" alt="" title=""><span>MAMMALS : A little bit of text can go here.</span></div></li>
-<li class="p3"><div><a class="a3" href="#p3"><b>BIRDS</b></a><img src="http://www.cssplay.co.uk/menus/horizontalaccordian/pic3.jpg" alt="" title=""><span>BIRDS : Some information about birds can go here.</span></div></li>
-<li class="p4"><div><a class="a4" href="#p4"><b>REPTILES</b></a><img src="http://www.cssplay.co.uk/menus/horizontalaccordian/pic4.jpg" alt="" title=""><span>REPTILES : The size of this panel can be controlled in the stylesheet.</span></div></li>
-<li class="p5"><div><a class="a5" href="#p5"><b>FISH</b></a><img src="http://www.cssplay.co.uk/menus/horizontalaccordian/pic5.jpg" alt="" title=""><span>FISH : Freshwater, tropical and marine fish.</span></div></li>
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-</html>'''Producing high-value proteins in ''Phaeodactylum tricornutum'' '''
-The diatom ''Phaeodactylum tricornutum'' is a widely spread organism in marine waters. Together with about 6000 other species, diatoms belong to the phylum of heterokonts. As a group of great ecological relevance diatoms are responsible for up to 20% of the global CO<sub>2</sub> fixation and generate about 40 % of the marine biomass of primary producers (Falkowski ''et al.'', 1998, Field ''et al.'', 1998).
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-In addition, diatoms represent an important source of lipids and silicate making them interesting for various biotechnological applications e.g. in biofuel industry, food industry and nanofabrication.
-“Normal” biotechnology is applied in bacteria and yeast.  Both need to be fed with cost intensive chemicals and the purification of the products e.g. Proteins is very time consuming and difficult, due to the fact that the cells need to be cracked or chemicals in the medium need to be removed.
-In contrast, ''P. tricornutum'' is a green organism living from CO<sub>2</sub> and light and love. The purification of the desired product is much easier, because the proteins are secreted to the surrounding medium. As a plant, microalgae don’t secret many other substances and therefore our product is already in a high purity available (Hempel ''et al.'', 2011).
-In order to introduce this organism as a chassis to the iGEM community, we build a toolbox of expression vectors, promoters, selection markers, reporter proteins and signal peptides for ''P. tricornutum''. This allows every future iGEM team to produce their own complex proteins, which can easily be purified.
-[[File:Phaeodactylum.jpg|500px|center]]
-<center>Foto: © Ansgar Gruber, University Konstanz</center>
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