Difference between revisions of "Team:TU Darmstadt/Sustainable"

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<br> <br>
 
<br> <br>
<i>B.&nbsp;subtilis</i> is a gram-positive bacterium which is usually found in soil.
+
<i>B.&nbsp;subtilis</i> is a <b>gram-positive bacterium</b> which is usually found in soil.
  Gram-positive bacteria only have one cell membrane and are thus often used for protein secretion<sup id="cite_ref-1"><a href="#cite_note-1">[1]</a></sup>.   
+
  Gram-positive bacteria only have <b>one cell membrane</b> and are thus often used for <b>protein secretion</b><sup id="cite_ref-1"><a href="#cite_note-1">[1]</a></sup>.   
  On the one hand, secreting proteins is beneficial in protein production because no cell disruption is
+
  On the one hand, secreting proteins is <b>beneficial in protein production</b> because no cell disruption is
 
   necessary to collect the protein and the formation of inclusion bodies can be prevented<sup id="cite_ref-2"><a href="#cite_note-2">[2]</a></sup>. On the
 
   necessary to collect the protein and the formation of inclusion bodies can be prevented<sup id="cite_ref-2"><a href="#cite_note-2">[2]</a></sup>. On the
   other hand, and in our case specifically, we avoid the issue of bringing enzyme substrates across
+
   other hand, and in our case specifically, we <b>avoid the issue of bringing enzyme substrates across
     the membrane in the cell as well as potential cell toxicity of the substrate and degradation product.
+
     the membrane in the cell</b> as well as <b>potential cell toxicity</b> of the substrate and degradation product.
 
     We want to leverage the ability of <i>B.&nbsp;subtilis</i> to secret proteins to immobilize our target enzymes
 
     We want to leverage the ability of <i>B.&nbsp;subtilis</i> to secret proteins to immobilize our target enzymes
 
       in the biofilm matrix by fusing them to a matrix protein. This is easier to realize with
 
       in the biofilm matrix by fusing them to a matrix protein. This is easier to realize with
 
       <i>B.&nbsp;subtilis</i> as gram-positive model organism than with the gram-negative bacteria,
 
       <i>B.&nbsp;subtilis</i> as gram-positive model organism than with the gram-negative bacteria,
         e.g. <i>Escherichia&nbsp;coli</i>. In addition, <i>B.&nbsp;subtilis</i> is a facultative anaerobic
+
         e.g. <i>Escherichia&nbsp;coli</i>. In addition, <i>B.&nbsp;subtilis</i> is a <b>facultative anaerobic
         organism which means it can grow in oxygenic, as well as anaerobic conditions,
+
         organism</b> which means it can grow in oxygenic, as well as anaerobic conditions,
 
           which can be of advantage when using the biofilm underwater in the clarifier<sup id="cite_ref-3"><a href="#cite_note-3">[3]</a></sup>.
 
           which can be of advantage when using the biofilm underwater in the clarifier<sup id="cite_ref-3"><a href="#cite_note-3">[3]</a></sup>.
  
Line 104: Line 104:
 
In nature <i>B.&nbsp;subtilis</i> forms biofilms on plant roots as it needs a nutrient source. The
 
In nature <i>B.&nbsp;subtilis</i> forms biofilms on plant roots as it needs a nutrient source. The
 
  advantage for the plants is that <i>B.&nbsp;subtilis</i> protects it from pathogenic infections by funghi<sup id="cite_ref-4"><a href="#cite_note-4">[4]</a></sup>.
 
  advantage for the plants is that <i>B.&nbsp;subtilis</i> protects it from pathogenic infections by funghi<sup id="cite_ref-4"><a href="#cite_note-4">[4]</a></sup>.
   Since <i>B.&nbsp;subtilis</i> is a natural biofilm former, it has been used as model organism  
+
   Since <i>B.&nbsp;subtilis</i> is a <b>natural biofilm former</b>, it has been used as <b>model organism  
   for studying biofilms in the last years<sup id="cite_ref-4"><a href="#cite_note-4">[4]</a></sup>. By using this natural biofilm former, we do not have to
+
   for studying biofilms</b> in the last years<sup id="cite_ref-4"><a href="#cite_note-4">[4]</a></sup>. By using this natural biofilm former, we do not have to
 
   worry about how our organism will form a biofilm. Furthermore, <i>B.&nbsp;subtilis</i> is classified as
 
   worry about how our organism will form a biofilm. Furthermore, <i>B.&nbsp;subtilis</i> is classified as
     generally recognized as safe (GRAS) by the American&nbsp;Food&nbsp;and&nbsp;Drug&nbsp;Administration&nbsp;(FDA)<sup id="cite_ref-5"><a href="#cite_note-5">[5]</a></sup>, unlike other
+
     <b>generally recognized as safe (GRAS)</b> by the American&nbsp;Food&nbsp;and&nbsp;Drug&nbsp;Administration&nbsp;(FDA)<sup id="cite_ref-5"><a href="#cite_note-5">[5]</a></sup>, unlike other
 
     bacteria which are often pathogenic like <i>E.&nbsp;coli</i> or <i>Staphylococcus&nbsp;aureus</i><sup id="cite_ref-6"><a href="#cite_note-6">[6]</a></sup>. It is very  
 
     bacteria which are often pathogenic like <i>E.&nbsp;coli</i> or <i>Staphylococcus&nbsp;aureus</i><sup id="cite_ref-6"><a href="#cite_note-6">[6]</a></sup>. It is very  
 
     important for us to use an organism which is generally safe given the fact that we want to bring it  
 
     important for us to use an organism which is generally safe given the fact that we want to bring it  
Line 114: Line 114:
  
 
<br> <br>
 
<br> <br>
Another characteristic of <i>B.&nbsp;subtilis</i> is its ability to form endospores
+
Another characteristic of <i>B.&nbsp;subtilis</i> is its <b>ability to form endospores</b>
  when exposed to extreme environmental conditions<sup id="cite_ref-7"><a href="#cite_note-7">[7]</a></sup>. Spores are highly resistant
+
  when exposed to extreme environmental conditions<sup id="cite_ref-7"><a href="#cite_note-7">[7]</a></sup>. Spores are highly <b>resistant
   towards extreme temperature, desiccation, radiation and chemicals. This  
+
   towards extreme temperature, desiccation, radiation and chemicals</b>. This  
 
   resistance makes spores interesting for industrial processes, e.g. by displaying  
 
   resistance makes spores interesting for industrial processes, e.g. by displaying  
 
   enzymes on the surface of spores which can then be used in harsh reaction conditions. Furthermore,  
 
   enzymes on the surface of spores which can then be used in harsh reaction conditions. Furthermore,  
   they are metabolic inactive but can germinate once nutrients are sufficiently available or when they  
+
   they are <b>metabolic inactive</b> but <b>can germinate</b> once nutrients are sufficiently available or when they  
 
   experience high pressure<sup id="cite_ref-8"><a href="#cite_note-8">[8]</a></sup>.
 
   experience high pressure<sup id="cite_ref-8"><a href="#cite_note-8">[8]</a></sup>.
  

Revision as of 09:52, 17 October 2020

image/svg+xml - O O



Biofilms

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At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. [1]

Bacillus subtilis

We aim to convert micropollutants in wastewater treatment plants using an enzyme-producing biofilm and came to the decision that Bacillus subtilis is the best choice as our biofilm forming organism.

B. subtilis is a gram-positive bacterium which is usually found in soil. Gram-positive bacteria only have one cell membrane and are thus often used for protein secretion[1]. On the one hand, secreting proteins is beneficial in protein production because no cell disruption is necessary to collect the protein and the formation of inclusion bodies can be prevented[2]. On the other hand, and in our case specifically, we avoid the issue of bringing enzyme substrates across the membrane in the cell as well as potential cell toxicity of the substrate and degradation product. We want to leverage the ability of B. subtilis to secret proteins to immobilize our target enzymes in the biofilm matrix by fusing them to a matrix protein. This is easier to realize with B. subtilis as gram-positive model organism than with the gram-negative bacteria, e.g. Escherichia coli. In addition, B. subtilis is a facultative anaerobic organism which means it can grow in oxygenic, as well as anaerobic conditions, which can be of advantage when using the biofilm underwater in the clarifier[3].

In nature B. subtilis forms biofilms on plant roots as it needs a nutrient source. The advantage for the plants is that B. subtilis protects it from pathogenic infections by funghi[4]. Since B. subtilis is a natural biofilm former, it has been used as model organism for studying biofilms in the last years[4]. By using this natural biofilm former, we do not have to worry about how our organism will form a biofilm. Furthermore, B. subtilis is classified as generally recognized as safe (GRAS) by the American Food and Drug Administration (FDA)[5], unlike other bacteria which are often pathogenic like E. coli or Staphylococcus aureus[6]. It is very important for us to use an organism which is generally safe given the fact that we want to bring it into a wastewater treatment plant.

Another characteristic of B. subtilis is its ability to form endospores when exposed to extreme environmental conditions[7]. Spores are highly resistant towards extreme temperature, desiccation, radiation and chemicals. This resistance makes spores interesting for industrial processes, e.g. by displaying enzymes on the surface of spores which can then be used in harsh reaction conditions. Furthermore, they are metabolic inactive but can germinate once nutrients are sufficiently available or when they experience high pressure[8].

Micropollutants in Wastewater

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Laccases

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EreB

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Modularity

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Quorum Sensing

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At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet.

Kill Switches

Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua.
At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet. Lorem ipsum dolor sit amet, consetetur sadipscing elitr, sed diam nonumy eirmod tempor invidunt ut labore et dolore magna aliquyam erat, sed diam voluptua. At vero eos et accusam et justo duo dolores et ea rebum. Stet clita kasd gubergren, no sea takimata sanctus est Lorem ipsum dolor sit amet.

References

1. Liu, L.; Liu, Y.; Shin, H. D. Developing Bacillus Spp. as a Cell Factory for Production of Microbial Enzymes and Industrially Important Biochemicals in the Context of Systems and Synthetic Biology. Applied Microbiology and Biotechnology. Springer July 11, 2013, pp 6113–6127 Doi:10.1007/s00253-013-4960-4 2.Westers, L.; Westers, H.; Quax, W. J. Bacillus Subtilis as Cell Factory for Pharmaceutical Proteins: A Biotechnological Approach to Optimize the Host Organism. Biochimica et Biophysica Acta - Molecular Cell Research. Elsevier November 11, 2004, pp 299–310 Doi:10.1016/j.bbamcr.2004.02.011 3.Clements, L. D.; Miller, B. S.; Streips, U. N. Comparative Growth Analysis of the Facultative Anaerobes Bacillus Subtilis, Bacillus Licheniformis, and Escherichia Coli. Syst. Appl. Microbiol. 2002, 25 (2), 284–286 Doi:10.1078/0723-2020-00108 4. Vlamakis, H.; Chai, Y.; Beauregard, P. Sticking Together: Building a Biofilm the Bacillus Subtilis Way. Nature Reviews Microbiology. NIH Public Access March 2013, pp 157–168 Doi:10.1038/nrmicro2960 5. Microorganisms & Microbial-Derived Ingredients Used in Food (Partial List) | FDA https://www.fda.gov/food/generally-recognized-safe-gras/microorganisms-microbial-derived-ingredients-used-food-partial-list (accessed Aug 27, 2020) 6.Hobley, L.; Harkins, C.; MacPhee, C. E. Giving Structure to the Biofilm Matrix: An Overview of Individual Strategies and Emerging Common Themes. FEMS Microbiology Reviews. Oxford University Press June 8, 2015, pp 649–669 Doi:10.1093/femsre/fuv015 7. Karava, M.​; Gockel, P.; Kabisch, J. Bacillus Subtilis​ Spore Surface Display of Photodecarboxylase for the Transformation of Lipids to Hydrocarbons. bioRxiv 2020, 2020.08.30.273821 Doi:10.1101/2020.08.30.273821 8.Black, E. P.; Koziol-Dube, K.; Guan, D. Factors Influencing Germination of Bacillus Subtilis Spores via Activation of Nutrient Receptors by High Pressure. Appl. Environ. Microbiol. 2005, 71 (10), 5879–5887 Doi:10.1128/AEM.71.10.5879-5887.2005