Part:BBa_K3588001

CocE, Rhodococcus sp. (strain MB1 Bresler)

This part is the CDS of the enzyme Cocaine Esterase (CocE) from Rhodococcus sp. (strain MB1 Bresler). This enzyme hydrolyses cocaine faster than other known natural cocaine esterase.

Sequence and Features

Assembly Compatibility:

• 10
  COMPATIBLE WITH RFC[10]
• 12
  COMPATIBLE WITH RFC[12]
• 21
  INCOMPATIBLE WITH RFC[21]

  Illegal BamHI site found at 1188
  
• 23
  COMPATIBLE WITH RFC[23]
• 25
  COMPATIBLE WITH RFC[25]
• 1000
  COMPATIBLE WITH RFC[1000]

Synthesis

In trying to order this from IDT the genetic code was too complex and so we ran codon optimisation, the code included has been optimised for E. coli.

Function

The enzyme CocE catalyses the breakdown of cocaine into benzoate and ecgonine methyl ester

The enzyme hydrolyzes cocaine to benzoate and ecgonine methyl ester, endowing the bacteria with the ability to utilize cocaine as a sole source of carbon and energy for growth, as this bacterium lives in the rhizosphere of coca plants. Also efficiently hydrolyzes cocaethylene, a more potent cocaine metabolite that has been observed in patients who concurrently abuse cocaine and alcohol. Is able to prevent cocaine-induced convulsions and lethality in rat. The bacterial strain this enzyme originates from the rhizosphere of the coca plant (the plant that is used to produce cocaine).

Properties

Cocaine Substrate Kinetics

k_cat is 7.8 sec^-1 K_M=0.64 µM

Cocaethylene Substrate Kinetics

k_cat is 9.4 sec^-1 K_M=1.6 µM

Other Details

Half Life (at 37°C = ~6 mins)

pH dependence on cocE kinetics and stability.