Difference between revisions of "Part:BBa K3588001"
 
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<partinfo>BBa_K3588001 short</partinfo>
 
<partinfo>BBa_K3588001 short</partinfo>
  
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<h1> hello </h1)
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This part is the CDS of the enzyme Cocaine Esterase (CocE) from <i>Rhodococcus sp. (strain MB1 Bresler)</i>. This enzyme hydrolyses cocaine faster than other known natural cocaine esterase.
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This part is the wildtype ezyme
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===Usage and Biology===
 
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<span class='h3bb'>Sequence and Features</span>
 
<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K3588001 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K3588001 SequenceAndFeatures</partinfo>
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===Synthesis===
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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.
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===Function===
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[[File:CocE Catalytic Activity.png|none|800px|thumb| The enzyme CocE catalyses the breakdown of cocaine into benzoate and ecgonine methyl ester <ref>http://dx.doi.org/10.1128/aem.66.3.904-908.2000</ref>]]
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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.<ref>https://faseb.onlinelibrary.wiley.com/doi/abs/10.1096/fasebj.5.12.1916095</ref> 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).<ref>https://aem.asm.org/content/66/3/904</ref>
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===Properties===
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<h5>Cocaine Substrate Kinetics</h5>
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k<sub>cat</sub> is 7.8 sec<sup>-1</sup>
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K<sub>M</sub>=0.64 µM<ref>http://dx.doi.org/10.1021/bi026131p</ref>
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<h5>Cocaethylene Substrate Kinetics</h5>
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k<sub>cat</sub> is 9.4 sec<sup>-1</sup>
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K<sub>M</sub>=1.6 µM<ref>http://dx.doi.org/10.1021/bi026131p</ref>
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<h5>Other Details</h5>
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Half Life (at 37°C = ~6 mins) <ref>https://pubs.acs.org/doi/10.1021/cb500257s</ref>
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[[File:CocE Caracterisation.png|none|400px|thumb| ''pH dependence on cocE kinetics and stability.''<ref>scihub.wikicn.top/10.1021/bi026131p</ref>]]
  
  

Latest revision as of 03:06, 28 October 2020


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 [1]


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.[2] 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).[3]

Properties

Cocaine Substrate Kinetics

kcat is 7.8 sec-1 KM=0.64 µM[4]

Cocaethylene Substrate Kinetics

kcat is 9.4 sec-1 KM=1.6 µM[5]

Other Details

Half Life (at 37°C = ~6 mins) [6]

pH dependence on cocE kinetics and stability.[7]


  1. ↑ http://dx.doi.org/10.1128/aem.66.3.904-908.2000
  2. ↑ https://faseb.onlinelibrary.wiley.com/doi/abs/10.1096/fasebj.5.12.1916095
  3. ↑ https://aem.asm.org/content/66/3/904
  4. ↑ http://dx.doi.org/10.1021/bi026131p
  5. ↑ http://dx.doi.org/10.1021/bi026131p
  6. ↑ https://pubs.acs.org/doi/10.1021/cb500257s
  7. ↑ scihub.wikicn.top/10.1021/bi026131p