Difference between revisions of "Team:TU Darmstadt/Project/Bio/Monomeres/Haconsaeure"

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<h2 align="center">Biotechnological production of itaconic acid in <em>Escherichia coli</em></h2>
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<h2>Biotechnological production of itaconic acid in <em>Escherichia coli</em></h2>
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<p>The compilation of our toolbox requires a molecule containing at least two carboxygroups for esterifications with polyalcohols.&nbsp; Furthermore it is important that one compartment of the polymer contains a crosslinking functionality which composes bonds with other strings under specific conditions. A molecule that complies with both requirements is the dicarbonacid itaconic acid, which has an ethylene group that preserves the cross-linking functionality by addition reactions under radicalic activation.</p>
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<p>Itaconic acid is already used as a Co-monomer for the synthesis of polyacrylates and vulcanized rubber as well as a basic module for biologically degradable Polymers that are for example used in packaging industry. The property to act as a Co-monomer with different other components has the consequence that itaconic acid is discussed as possible substitute for methacrylic acid that is utilized to date by petrochemical industry.(1)</p>
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<p>The conventional exploitation of itaconic acid is achieved by chemical synthesis or takes place in <em>Aspergillus terreus</em> where amounts of 80g/L are provided.(2)An important disadvantage of the recent production is the high costs of approximately US$4/kg. <em>(3)</em></p>
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<p>A possibility to achieve a better production of itaconic acid in a different host than Aspergillus terreus could be performed by the introduction of a single key enzyme of the Aspergillus pathway.</p>
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<p>The cis-aconitate decarboxylase from Aspergillus terreus catalyzes the reaction of cis-aconitate to itaconic acid under loss of carbon dioxide. cis-aconitate is an intermediate of the TCA-cycle that occurs in most organisms. For that reason there are a lot of different possible hosts. We decided to use Escherichia coli which contains the whole TCA-cycle and is also well characterized.</p>
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<p>The compilation of our toolbox requires beside the implemented polyalcohols (xylitol and ethyleneglycol) a molecule containing two carboxygroups for esterifications with these polyalcohols.&nbsp; Furthermore it is important that one compartment of the polymer contains a crosslinking group which composes bonds with other strings under specific conditions. A molecule that complies with our requirements is the dicarbonacid itaconic acid. We, the iGEM team TU Darmstadt 2015; want to accomplish an alternative way of itaconic acid fabrication in <em>Escherichia coli</em> as host.</p>
 
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<p>Itaconic acid is already used as a Co-monomer for the synthesis of polyacrylates and vulcanized rubber as well as a basic module for biologically degradable Polymers that are for example used in packaging industry. The conventional exploitation of itaconic acid is achieved by chemical synthesis or takes place in <em>Aspergillus terreus</em> where amounts of 80g/L are provided.(1) An important disadvantage of the recent production is the high costs of approximately US$4/kg. <em>(2)</em> Along with further biotechnologically relevant chemicals, itaconic acid was listed by the U.S Department of Energy as one of the twelve platform chemicals with especially biotechnologically production potential. These wide areas of application as well as the existing opportunities for improvement are the reasons for us to focus partly on itaconic acid.(3)</p>
 
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<p>Itaconic acid has several chemically plus physico-chemical characteristics that we are going to use for us. The carboxyl groups allow itaconic acid to be esterified with hydroxyl-groups of several sugaralcohols, for example ethylene glycol, and thereby compose long chains of heteropolymers. The property to act as a Co-monomer with different other components has the consequence that itaconic acid is discussed as possible substitute for methacrylic acid that is utilized to date by petrochemical industry.(4)</p>
 
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<p>Another remarkable feature itaconic acid contains are the three different protonation states with p<sub>Ka</sub>-values of 3.83 and 5.55 (<em>(5)</em>;) that are typically for a dicarboxylic acid. Thereby the reaction responsiveness can be varied by adjusting the pH-value.</p>
 
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<p>To permute the idea of biotechnological production of itaconic acid in a different host than <em>Aspergillus terreus</em>, the required enzyme needs to be implemented in the host of our choice. Therefore we benefit from the already existing TCA-cycle intermediates metabolized by E.coli and tie in with the synthesis of cis-aconitate.<br /> The in <em>E.coli</em> metabolism newly implemented Protein, cis-aconitate-decarboxylase coded by <em>cadA</em>, forms cis-aconitate to itaconic acid.&nbsp;</p>
 
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<p>&nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp; &nbsp;<img src="filemanager/source/Itaconic acid pathway.jpg" alt="Itaconic acid pathway" width="392" height="439" /></p>
 
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<li>Huang X, Lu X, Li Y, Li X, Li JJ. Improving itaconic acid production through genetic engineering of an industrial Aspergillus terreus strain. Microb Cell Fact. 2014;13:119.</li>
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lang=EN-US><span style='mso-element:field-begin'></span><span
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<li>Huang X, Chen M, Lu X, Li Y, Li X, Li JJ. Direct production of itaconic acid from liquefied corn starch by genetically engineered Aspergillus terreus. Microb Cell Fact. 2014;13:108.</li>
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<li>Willke T, Vorlop KD. Biotechnological production of itaconic acid. Appl Microbiol Biotechnol. 2001;56(3-4):289-95.</li>
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field-separator'></span></span><![endif]--><span lang="EN-US">1.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Huang X, Chen M, Lu X, Li Y, Li X, Li JJ. Direct production of itaconic acid from liquefied corn starch by genetically engineered Aspergillus terreus. Microb Cell Fact. 2014;13:108.</span></p>
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<p class="EndNoteBibliography"><span lang="EN-US">2.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Willke T, Vorlop KD. Biotechnological production of itaconic acid. Appl Microbiol Biotechnol. 2001;56(3-4):289-95.</span></p>
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<p class="EndNoteBibliography"><span lang="EN-US">3.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Okabe M, Lies D, Kanamasa S, Park EY. Biotechnological production of itaconic acid and its biosynthesis in Aspergillus terreus. Appl Microbiol Biotechnol. 2009;84(4):597-606.</span></p>
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<p class="EndNoteBibliography"><span lang="EN-US">4.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Huang X, Lu X, Li Y, Li X, Li JJ. Improving itaconic acid production through genetic engineering of an industrial Aspergillus terreus strain. Microb Cell Fact. 2014;13:119.</span></p>
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<p class="EndNoteBibliography"><span lang="EN-US">5.&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp;&nbsp; Klement T, Buchs J. Itaconic acid--a biotechnological process in change. Bioresour Technol. 2013;135:422-31.</span></p>
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Revision as of 14:27, 16 September 2015

Biotechnological production of itaconic acid in Escherichia coli

 

The compilation of our toolbox requires a molecule containing at least two carboxygroups for esterifications with polyalcohols.  Furthermore it is important that one compartment of the polymer contains a crosslinking functionality which composes bonds with other strings under specific conditions. A molecule that complies with both requirements is the dicarbonacid itaconic acid, which has an ethylene group that preserves the cross-linking functionality by addition reactions under radicalic activation.

Itaconic acid is already used as a Co-monomer for the synthesis of polyacrylates and vulcanized rubber as well as a basic module for biologically degradable Polymers that are for example used in packaging industry. The property to act as a Co-monomer with different other components has the consequence that itaconic acid is discussed as possible substitute for methacrylic acid that is utilized to date by petrochemical industry.(1)

The conventional exploitation of itaconic acid is achieved by chemical synthesis or takes place in Aspergillus terreus where amounts of 80g/L are provided.(2)An important disadvantage of the recent production is the high costs of approximately US$4/kg. (3)

A possibility to achieve a better production of itaconic acid in a different host than Aspergillus terreus could be performed by the introduction of a single key enzyme of the Aspergillus pathway.

The cis-aconitate decarboxylase from Aspergillus terreus catalyzes the reaction of cis-aconitate to itaconic acid under loss of carbon dioxide. cis-aconitate is an intermediate of the TCA-cycle that occurs in most organisms. For that reason there are a lot of different possible hosts. We decided to use Escherichia coli which contains the whole TCA-cycle and is also well characterized.

 

 

 

 

  1. Huang X, Lu X, Li Y, Li X, Li JJ. Improving itaconic acid production through genetic engineering of an industrial Aspergillus terreus strain. Microb Cell Fact. 2014;13:119.
  2. Huang X, Chen M, Lu X, Li Y, Li X, Li JJ. Direct production of itaconic acid from liquefied corn starch by genetically engineered Aspergillus terreus. Microb Cell Fact. 2014;13:108.
  3. Willke T, Vorlop KD. Biotechnological production of itaconic acid. Appl Microbiol Biotechnol. 2001;56(3-4):289-95.