Part:BBa_K3215002

phnG optimized

This part provides a codon optimized phnG CDS from E. coli K-12, since its translation efficiency can be improved in relation to the natural CDS.

Usage and Biology

The phn Operon is one of the nine transcriptional units that are present in the Phosphate Regulon (Pho), a global regulatory mechanism involved in bacterial Pi management. It is responsible for the catabolism of phosphates and it has 14 cistrons (phnCDEFGHIJKLMNOP) that, together, are responsible for controlling the Carbon-Phosphorus (C-P) lyase activity. The C-P lyase pathway is activated upon conditions of phosphate starvation and it is responsible for converting phosphonates into 5-phosphoribosyl-α-1-diphosphate (PRPP), utilizing ATP.

phnGHIJKLM

It forms the C-P lyase core complex and is responsible for the coupling of phosphonate to ATP and C-P bond cleavage. In this complex, the PhnI, a nucleosidase capable of deglycosylating ATP and GTP to ribose 5-triphosphate, is supported by phnG, phnH, and phnL in the catalysis of the transfer of phosphonate moiety to a 5′-triphosphate alkyl phosphonate intermediate, according to the following reaction:

After that, PhnM catalyses the pyrophosphate release through the reaction:

Following, phnJ is allowed to cleave the C-P lyase bond via an S-adenosyl methionine (SAM)-dependent radical mechanism, thus converting the resulting ribose cyclic phosphate into PRPP. This reaction is possible because of the interaction of phnK trough the via its α helices 3 and 4 with PhnJ, exposing its active site.

Sequence and Features

Assembly Compatibility:

• 10
  INCOMPATIBLE WITH RFC[10]

  Illegal PstI site found at 209
  
• 12
  INCOMPATIBLE WITH RFC[12]

  Illegal PstI site found at 209
  
• 21
  COMPATIBLE WITH RFC[21]
• 23
  INCOMPATIBLE WITH RFC[23]

  Illegal PstI site found at 209
  
• 25
  INCOMPATIBLE WITH RFC[25]

  Illegal PstI site found at 209
  
• 1000
  COMPATIBLE WITH RFC[1000]