Part:BBa_K3349000

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.

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]