Part:BBa_K4763007

AtPCS (phytochelatin synthase) coding sequence

Phytochelatin synthase (AtPCS) coding sequence from Arabidopsis thaliana. This enzyme catalyzes the final step in the biosynthesis of phytochelatins (PCs).

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal XbaI site found at 649
  
• 12
  COMPATIBLE WITH RFC[12]
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal XbaI site found at 649
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal XbaI site found at 649
  
• 1000
  COMPATIBLE WITH RFC[1000]

Usage and Biology

The enzyme chosen for the generation of the biopart was phytochelatin synthase or PCS (EC 2.3.2.15), which catalyzes the synthesis of glutathione (GSH) polymers called phytochelatins (PCs) (Figure 1). These are produced as a mechanism of resistance and accumulation in algae, yeast, plants, and worms in response to heavy metal stress. Phytochelatins bind to metals intracellularly and render them inactive. This is because the substrate of PCS is GSH (as aforementioned) and other metal-bis-glutathione complexes. Additionally, the enzyme activity requires the presence of specific heavy metals (depending on the species under study), with cadmium being the main inducer in plants and other organisms. The complexes formed by the metal and PCs can be subsequently compartmentalized into vacuoles, chloroplasts, and mitochondria (García-García et al., 2014; 2020).

The PCS enzyme from the model organism Arabidopsis thaliana (AtPCS1) was selected for the creation of this biopart due to its more extensive characterization in the synthesis of phytochelatins.

File:Phytochelatin synthesis diagram.png

Figure 1. Phytochelatin synthesis from GSH, catalyzed by PCS.

Characterization

Cloning into pET28(+)

Colony PCR

Digestion

Protein overexpression