Composite

Part:BBa_K3349009

Designed by: Kristi Turton   Group: iGEM20_Lethbridge_HS   (2020-09-28)


Pectin Lyase Pnl-GFP

This Pectin lyase is originally from Paenabacillus amylolyticus. Pnl cleaves at methylated sites of pectin or homogalacturonan substrates at high affinity [1]. Used in conjunction 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 a pH of 9-10 [1]. For more information, please visit our website https://2020.igem.org/Team:Lethbridge_HS or more analysis of the pnl gene see part BBa_K3349000.

A GFP reporter has been fused via a GC linker to the C terminal end of the protein. As well a C-terminal his tag was also added.

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]


[edit]
Categories
Parameters
None