Difference between revisions of "Part:BBa K3349002"
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The PelC Pectin Lyase is originally from <i> Paenabacillus amylolyticus </i>. This particular pectin lyase is more specific to methylated pectin or homogalacturonan substrates rather than polygalacturonic acid [1]. This protein also works with both PelB(BBa_K3349001) and Pnl(BBa_K3349001) in this organism to effectively break down pectin. PelC has an optimal activity at 55°C and a pH of 10. The Km in in terms of 80% methylated substrates is 0.41 +/-0.06mg/ml. Calcium is a cofactor for this protein and is therefore necessary within the buffer conditions [1].   
 
The PelC Pectin Lyase is originally from <i> Paenabacillus amylolyticus </i>. This particular pectin lyase is more specific to methylated pectin or homogalacturonan substrates rather than polygalacturonic acid [1]. This protein also works with both PelB(BBa_K3349001) and Pnl(BBa_K3349001) in this organism to effectively break down pectin. PelC has an optimal activity at 55°C and a pH of 10. The Km in in terms of 80% methylated substrates is 0.41 +/-0.06mg/ml. Calcium is a cofactor for this protein and is therefore necessary within the buffer conditions [1].   
  
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For purification purposes, this gene has a C-terminal hexa-histidine tag. For more information on our design and use please see our webpage https://2020.igem.org/Team:Lethbridge_HS.  
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For purification purposes, this gene has a C-terminal hexa-histidine tag. For more information on our design and use please see our webpage https://2020.igem.org/Team:Lethbridge_HS.
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In the beginning of our project we decided to do homology modelling of our pectin lyase enzymes in order to inform our thermostability modelling.
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<img src= "https://2020.igem.org/wiki/images/2/22/T--Lethbridge_HS--PelCmodelling.png" alt="pnl modelling" style="width:700px;height:300px;">
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</html>
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<b>Figure 1:</b> Homology modelling of Pnl from <i>P. amylolyticus </i>. The protein structure was done using iTASSER [2] and SWISS-Model [3]. The two structures were then aligned and compared. A more in depth analysis can be found on our webpage.
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===Usage and Biology===
 
===Usage and Biology===

Revision as of 22:19, 1 October 2020


Pectin Lyase PelC

The PelC Pectin Lyase is originally from Paenabacillus amylolyticus . This particular pectin lyase is more specific to methylated pectin or homogalacturonan substrates rather than polygalacturonic acid [1]. This protein also works with both PelB(BBa_K3349001) and Pnl(BBa_K3349001) in this organism to effectively break down pectin. PelC has an optimal activity at 55°C and a pH of 10. The Km in in terms of 80% methylated substrates is 0.41 +/-0.06mg/ml. Calcium is a cofactor for this protein and is therefore necessary within the buffer conditions [1].

For purification purposes, this gene has a C-terminal hexa-histidine tag. For more information on our design and use please see our webpage https://2020.igem.org/Team:Lethbridge_HS.


In the beginning of our project we decided to do homology modelling of our pectin lyase enzymes in order to inform our thermostability modelling.


pnl modelling

Figure 1: Homology modelling of Pnl from P. amylolyticus . The protein structure was done using iTASSER [2] and SWISS-Model [3]. The two structures were then aligned and compared. A more in depth analysis can be found on our webpage.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    COMPATIBLE WITH RFC[21]
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal AgeI site found at 829
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI.rc site found at 215