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 | + | 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.
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
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]