Difference between revisions of "Part:BBa K3349000"

 
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<partinfo>BBa_K3349000 short</partinfo>
 
<partinfo>BBa_K3349000 short</partinfo>
  
This Pectin Lyase is from the organism <i> Paenibacillus amylolyticus </i>. Pnl cleaves at methylated sites of pectin or homogalacturonan substrates at high affinity [1]. Used in conjuction with Pectin lyases PelB and PelC, pectin can be effectively cleaved [1]. A hexa-histidine tag was added to the C-terminus of the coding sequence for nickel affinity purification.  
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This Pectin Lyase is from the organism <i> Paenibacillus amylolyticus </i>. Pnl cleaves at methylated sites of pectin or homogalacturonan substrates at high affinity [1]. Used in conjuction with Pectin lyases PelB and PelC, pectin can be completely degraded [1]. Keggi and Doran-Peterson found that this pectinase had a Km of 0.13 mg/ml and a Vmax of 144.6+/- 1.894 iU/mg when a 80% methylated substrate was used. Optimal reaction temperature was found to be 55°C and optimal pH between 9-10 [1].
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A hexa-histidine tag was added to the C-terminus of the coding sequence for nickel affinity purification. For more information, please visit our website https://2020.igem.org/Team:Lethbridge_HS
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For building thermostable variants of this Pnl enzyme we first did homology modelling as the crystal structure has not been determined.
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<img src= "https://2020.igem.org/wiki/images/c/c0/T--Lethbridge_HS--Pnl_modelling.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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<!-- Add more about the biology of this part here
 
<!-- Add more about the biology of this part here
 
===Usage and Biology===
 
===Usage and Biology===
 
For building thermostable variants of this Pnl enzyme we first did some homology modelling as the crystal structure has not been determined.
 
  
  
[T--Lethbridge_HS--Pnl modelling.png]
 
  
  

Latest revision as of 15:02, 7 October 2020


Pectin Lyase Pnl


This Pectin Lyase is from the organism Paenibacillus amylolyticus . Pnl cleaves at methylated sites of pectin or homogalacturonan substrates at high affinity [1]. Used in conjuction with Pectin lyases PelB and PelC, pectin can be completely degraded [1]. Keggi and Doran-Peterson found that this pectinase had a Km of 0.13 mg/ml and a Vmax of 144.6+/- 1.894 iU/mg when a 80% methylated substrate was used. Optimal reaction temperature was found to be 55°C and optimal pH between 9-10 [1].

A hexa-histidine tag was added to the C-terminus of the coding sequence for nickel affinity purification. For more information, please visit our website https://2020.igem.org/Team:Lethbridge_HS

For building thermostable variants of this Pnl enzyme we first did homology modelling as the crystal structure has not been determined.

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
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]