Difference between revisions of "Part:BBa K3349000"
Kristiturton (Talk | contribs) |
Kristiturton (Talk | contribs) |
||
| (8 intermediate revisions by the same user not shown) | |||
| Line 3: | Line 3: | ||
<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 | + | |
| + | 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]. | ||
| + | |||
| + | 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. | ||
| + | |||
| + | <html> | ||
| + | <img src= "https://2020.igem.org/wiki/images/c/c0/T--Lethbridge_HS--Pnl_modelling.png" alt="pnl modelling" style="width:700px;height:300px;"> | ||
| + | |||
| + | </html> | ||
| + | |||
| + | <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. | ||
| + | |||
<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
===Usage and Biology=== | ===Usage and Biology=== | ||
| − | |||
| − | |||
| − | |||
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.
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]