Difference between revisions of "Part:BBa K3349007"

Revision as of 15:16, 7 October 2020

Pectin Lyase PelB

This Pectin Lyase is a protein from Paenibaciullus amylolyticus. With Pnl and PelC, PelB is able to cleave and therefore breakdown pectin or homogalacturonan substrates. This protein has been characterized to cleave methylated pectin (at unmethylated sites) and polygalacturonic acid [2]. Studies by Keggi and Doran-Peterson, PelB has a Km of 50.1+/- 140 mg/ml and a Vmax of 745 +/-2024 IU/mg with a 80% methylated substrate. The optimal temperature for PelB activity was determined to be 70°C [1].

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

Although this enzyme is already fairly thermostable we decided to do homology modelling if we were to apply our modelling protocol to make a new PelB thermostable variant.

pnl modelling

Figure 2: Homology modelling of PelB 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
INCOMPATIBLE WITH RFC[21]
Illegal BglII site found at 333
Illegal BamHI site found at 1218
23
COMPATIBLE WITH RFC[23]
25
COMPATIBLE WITH RFC[25]
1000
COMPATIBLE WITH RFC[1000]

@@ Line 3: / Line 3: @@
 <partinfo>BBa_K3349007 short</partinfo>
-This Pectin Lyase is a hypothetical protein from <i> Paenibaciullus amylolyticus </i>. With Pnl and PelC, PelB is able to cleave and therefore breakdown pectin or homogalacturonan substrates. This protein has been characterized to cleave methylated pectin (at unmethylated sites) and polygalacturonic acid [2]. Studies by Keggi and Doran-Peterson, PelB has a Km of 50.1+/- 140 mg/ml and a Vmax of 745 +/-2024 IU/mg with a 80% methylated substrate (0 and 55% methylation was also studied). The optimal temperature for PelB activity was determined to be 70°C [1].
+This Pectin Lyase is a protein from <i> Paenibaciullus amylolyticus</i>. With Pnl and PelC, PelB is able to cleave and therefore breakdown pectin or homogalacturonan substrates. This protein has been characterized to cleave methylated pectin (at unmethylated sites) and polygalacturonic acid [2]. Studies by Keggi and Doran-Peterson, PelB has a Km of 50.1+/- 140 mg/ml and a Vmax of 745 +/-2024 IU/mg with a 80% methylated substrate. The optimal temperature for PelB activity was determined to be 70°C [1].
-A hexahistidine tag was added onto the C-terminus for nickel affinity purification. For more information, please visit our website https://2020.igem.org/Team:Lethbridge_HS.
+A hexa-histidine tag was added onto the C-terminus for nickel affinity purification. For more information, please visit our website https://2020.igem.org/Team:Lethbridge_HS.
 Although this enzyme is already fairly thermostable we decided to do homology modelling if we were to apply our modelling protocol to make a new PelB thermostable variant.
@@ Line 15: / Line 15: @@
 </html>
-<b>Figure 2:</b> Homology modelling of PelB 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.
+<b>Figure 2:</b> Homology modelling of PelB 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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