Team:BGU ISRAEL/Basic Part


Basic Part

Protocatechuate Degradation Pathway

Pseudomonas putida is a bacterium capable of utilizing protocatechuate, a toxic molecule for most bacteria. The protocatechuate degradation pathway converts protocatechuate into 3-oxoadipate, which is further metabolized by the bacteria and enters the TCA cycle.
We chose to submit the genes encoding the enzymes in this pathway. We also characterized the utilization of protocatechuate by bacterium using M9 minimal medium with protocatechuate as a sole carbon source. We have not been able to submit the pcaB gene, encoding the 3-Carboxymuconate Cycloisomerase, since the sequence contained 4 PstI restriction cut sites, which we did not have time to alter using point mutations.

Part Name Type Description Designer Length
BBa_K2091000 Coding Protocatechuate 3,4-dioxygenase Alpha Subunit B. Vaknin 606
BBa_K2091001 Coding Protocatechuate 3,4-dioxygenase Beta Subunit B. Vaknin 720
BBa_K2091002 Coding 4-Carboxymuconolactone Decarboxylase B. Vaknin 393
BBa_K2091003 Coding 3-Oxoadipate enol-lactonase B. Vaknin 792

LC-Cutinase Protein Variants

LC-cutinase is an enzyme found to have the ability to degrade PET. We have chosen to improve this enzyme using rational mutagenesis. 4 mutant variants of the LC-cutinase protein were produced using the PROSS algorithm and one Codon-optimized version of the protein was designed using the Genome Compiler software.
We have tested and characterized these proteins using kinetic trials, bacterial growth assays and scanning electron microscopy imaging.

Part Name Type Description Designer Length
BBa_K2091004 Coding LC-Cutinase Codon Optimized I. Bariah, E. Zajfman, I. Segal 948
BBa_K2091005 Coding LC-Cutinase Variant F4 I. Bariah, E. Zajfman, I. Segal 948
BBa_K2091006 Coding LC-Cutinase Variant F7 I. Bariah, E. Zajfman, I. Segal 948
BBa_K2091007 Coding LC-Cutinase Variant R4 I. Bariah, E. Zajfman, I. Segal 948
BBa_K2091008 Coding LC-Cutinase Variant R7 I. Bariah, E. Zajfman, I. Segal 948


Ben-Gurion University of the Negev
Ben Gurion 1, Beer Sheva 8410501, Israel


Connect With Us!