Difference between revisions of "Part:BBa K3349000"

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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 effectively cleaved [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
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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 effectively cleaved [1]. Keggi and Doran-Peterson found 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 between a pH of 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
  
 
For building thermostable variants of this Pnl enzyme we first did homology modelling as the crystal structure has not been determined.  
 
For building thermostable variants of this Pnl enzyme we first did homology modelling as the crystal structure has not been determined.  

Revision as of 22:41, 30 September 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 effectively cleaved [1]. Keggi and Doran-Peterson found 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 between a pH of 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]