Difference between revisions of "Team:Stuttgart/Design"

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    <h1>LIGHT UP THE PIPE</h1>
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<h1> LIGHT UP THE PIPE - IV Parts for a better flow </h1>
  
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<h2>Part I - Keratinases</h2>
  
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<h2>Keratinases</h2>
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<p>Blablabla</p>
  
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    <p>Blablabla</p>
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BILD
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<h2>Part II - Esterases</h2>
  
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<p>In our project we used esterases for degrading the layer of grease and waxes on hair which increases the accessibility for the keratinases. We investigated two different esterases for their enzyme activity. One esterase from the registry (EstCS2 BBa_K1149002) and one esterase (LipB) supplied by Dr. Eggert from Evoxx were compared.  <p>
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<h2>Esterases</h2>
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    <p> In our project we used esterases for degrading the layer of grease and waxes on hair which increases the accessibility for the keratinases. We investigated two different esterases for their enzyme activity. One esterase from the registry (EstCS2 BBa_K1149002) and one esterase (LipB) supplied by Dr. Eggert from Evoxx were compared.  <p>
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<h3>EstCS2</h3>
 
<h3>EstCS2</h3>
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<h2>Lipases</h2>
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<h2>Part III - Lipases</h2>
  
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    <p>Blablabla</p>
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<p>Blablabla</p>
  
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<h2>Insertion of fragrances</h2>
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<h2>Part IV - The rose fragrance </h2>
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    <p>The microbial synthesis of natural flavor compounds has become a very attractive alternative to the chemical production.1 In recent years microorganisms such as E.coli and Yeast have been metabolically engineered to produce different flavors like limonene, geraniol or rose.1,2,3 For our project we discussed different approaches and choose two different scents: limonene and rose. </p>
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<h3>The limonene scent</h3>
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<p>The microbial synthesis of natural flavor compounds has become a very attractive alternative to the chemical production. In recent years microorganisms such as E.coli and Yeast have been metabolically engineered to produce different flavors like limonene, geraniol or rose. For our project we discussed different approaches and choose two different scents: limonene and rose. As special fragrance we want to install a lovely scent of rose in our microbial system. Hair are commonly made of Keratin (90%) and small amounts of amino acids, such as L-phenylalanine. This amino acid can be used as substrate for the production of 2-Phenylethylacetate (2-PEAc), which has a rose-like odor. Therefor this odor can act as an indicator for keratin degradation.
 +
In recent studies from Guo et al the 2-PEAc biosynthetic pathway was successfully designed and expressed in E.coli.1 This pathway comprised four steps (Fig.1): Aminotransferase (ARO 8) for transamination of L-phenylalanine to phenylpyruvate, 2-keto acid decarboxylase KDC for the decarboxylation of the phenylpyruvate to phenylacetaldehyde, aldehyde reductase YjgB for the reduction of phenylacetaldehyde to 2-Phenylethanol, alcohol acetyltransferase ATF1 for the esterification of 2-PE to 2-PEAc.</p>
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<h2>Part IV.2 The limonene fragrance </h2>
  
 
<p>Limonene is a well-known cyclic monoterpene which can occur in two optical forms.2 (D)-Limonene is one of the most important and widespread terpenes in the flavor and fragrance industry, for example in citrus-flavored products such as soft drinks and candy.2 The (L)-Limonene form has a more harsh turpentine-like odor with a lemon-note.2 For our project we choose an enzyme-cascade, beginning with acetyl-coA and leading to the product (L)-limonene. This biosynthetic pathway was designed and inserted in E.coli.</p>
 
<p>Limonene is a well-known cyclic monoterpene which can occur in two optical forms.2 (D)-Limonene is one of the most important and widespread terpenes in the flavor and fragrance industry, for example in citrus-flavored products such as soft drinks and candy.2 The (L)-Limonene form has a more harsh turpentine-like odor with a lemon-note.2 For our project we choose an enzyme-cascade, beginning with acetyl-coA and leading to the product (L)-limonene. This biosynthetic pathway was designed and inserted in E.coli.</p>
 
<br>
 
<br>
 +
</table>
  
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<h3>The rose scent</h3>
 
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<p>As second fragrance we want to install a lovely scent of rose in our microbial system. Hair are commonly made of Keratin (90%) and small amounts of amino acids, such as L-phenylalanine. This amino acid can be used as substrate for the production of 2-Phenylethylacetate (2-PEAc), which has a rose-like odor.1 Therefor this odor can act as an indicator for keratin degradation.
 
−
In recent studies from Guo et al the 2-PEAc biosynthetic pathway was successfully designed and expressed in E.coli.1 This pathway comprised four steps (Fig.1): Aminotransferase (ARO 8) for transamination of L-phenylalanine to phenylpyruvate, 2-keto acid decarboxylase KDC for the decarboxylation of the phenylpyruvate to phenylacetaldehyde, aldehyde reductase YjgB for the reduction of phenylacetaldehyde to 2-Phenylethanol, alcohol acetyltransferase ATF1 for the esterification of 2-PE to 2-PEAc.</p>
 
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    <img src=https://static.igem.org/mediawiki/2017/0/02/Pathwayrose.png />
 
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(1)Metabolic engineering of Escherichia coli for production of 2-Phenylethylacetate from L-phenylalanine (2017). Daoyi Guo, Lihua Zhang, Hong Pan and Xun Li.
 
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(2) Biotechnological production of limonene in microorganisms (2016). Esmer Jongedijk, Katarina Cankar, Markus Buchhaupt, Jens Schrader, Harro Bouwmeester and Jules Beekwilder.
 
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(3)Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli (2015). Wei Liu, Rubing Zhang, Ning Tian, Xin Xu, Yujing Cao, Mo Xian and Huizhou Liu.
 
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(4)Rose Scent: Genomics Approach to Discovering Novel Floral Fragrance–Related Genes (2002). Inna Guterman, Moshe Shalit, Naama Menda, Dan Piestun, Mery Dafny-Yelin, Gil Shalev, Einat Bar, Olga Davydov, Mariana Ovadis, Michal Emanuel, Jihong Wang, Zach Adam, Eran Pichersky, Efraim Lewinsohn, Dani Zamir, Alexander Vainstein, and David Weiss.
 
  
  
 +
<foot>
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(1) Metabolic engineering of Escherichia coli for production of 2-Phenylethylacetate from L-phenylalanine (2017). D. Guo and L. Zhang et. al.
 +
(2) Biotechnological production of limonene in microorganisms (2016). E. Jongedijk and K. Cankar et. al.
 +
(3) Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli (2015). W. Liu and R. Zhang et. al.
 +
(4) Rose Scent: Genomics Approach to Discovering Novel Floral Fragrance–Related Genes (2002). I. Guterman and M. Shalit et. al.
 +
</foot>
 
</div>
 
</div>
  

Revision as of 19:28, 13 October 2017

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JUDGING FORM

LIGHT UP THE PIPE - IV Parts for a better flow

Part I - Keratinases

Blablabla

 BILD

Part II - Esterases

In our project we used esterases for degrading the layer of grease and waxes on hair which increases the accessibility for the keratinases. We investigated two different esterases for their enzyme activity. One esterase from the registry (EstCS2 BBa_K1149002) and one esterase (LipB) supplied by Dr. Eggert from Evoxx were compared.


EstCS2

EstCS2 from the iGEM Imperial College 2013 was proved to be active. In their project the cells expressing these construct were grown and lysed by sonication and were utilized in a colourimetric assay with the substrate analog para-Nitrophenyl butyrate. In our project we didn’t purify the esterases but used the supernatant for the enzyme activity assay.


LipB

EstCS2 from the iGEM Imperial College 2013 was proved to be active. LipB showed an enzyme activity of 2,8 U/mL in the supernatant. First, we repeated the enzyme activity assay from the iGEM TU Darmstadt 2012 to determine the esterase with the highest enzyme activity.


 BILD

Part III - Lipases

Blablabla

Part IV - The rose fragrance

The microbial synthesis of natural flavor compounds has become a very attractive alternative to the chemical production. In recent years microorganisms such as E.coli and Yeast have been metabolically engineered to produce different flavors like limonene, geraniol or rose. For our project we discussed different approaches and choose two different scents: limonene and rose. As special fragrance we want to install a lovely scent of rose in our microbial system. Hair are commonly made of Keratin (90%) and small amounts of amino acids, such as L-phenylalanine. This amino acid can be used as substrate for the production of 2-Phenylethylacetate (2-PEAc), which has a rose-like odor. Therefor this odor can act as an indicator for keratin degradation. In recent studies from Guo et al the 2-PEAc biosynthetic pathway was successfully designed and expressed in E.coli.1 This pathway comprised four steps (Fig.1): Aminotransferase (ARO 8) for transamination of L-phenylalanine to phenylpyruvate, 2-keto acid decarboxylase KDC for the decarboxylation of the phenylpyruvate to phenylacetaldehyde, aldehyde reductase YjgB for the reduction of phenylacetaldehyde to 2-Phenylethanol, alcohol acetyltransferase ATF1 for the esterification of 2-PE to 2-PEAc.



 BILD

Part IV.2 The limonene fragrance

Limonene is a well-known cyclic monoterpene which can occur in two optical forms.2 (D)-Limonene is one of the most important and widespread terpenes in the flavor and fragrance industry, for example in citrus-flavored products such as soft drinks and candy.2 The (L)-Limonene form has a more harsh turpentine-like odor with a lemon-note.2 For our project we choose an enzyme-cascade, beginning with acetyl-coA and leading to the product (L)-limonene. This biosynthetic pathway was designed and inserted in E.coli.


(1) Metabolic engineering of Escherichia coli for production of 2-Phenylethylacetate from L-phenylalanine (2017). D. Guo and L. Zhang et. al. (2) Biotechnological production of limonene in microorganisms (2016). E. Jongedijk and K. Cankar et. al. (3) Utilization of alkaline phosphatase PhoA in the bioproduction of geraniol by metabolically engineered Escherichia coli (2015). W. Liu and R. Zhang et. al. (4) Rose Scent: Genomics Approach to Discovering Novel Floral Fragrance–Related Genes (2002). I. Guterman and M. Shalit et. al.