Difference between revisions of "Part:BBa K2242920"

 
 
Line 1: Line 1:
  
 +
__NOTOC__
 +
<partinfo>BBa_K2242920 short</partinfo>
 +
 +
Ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE] is a key intermediate for the production of chiral drugs, including choles- terol-lowering HMG-CoA reductase inhibitors such as Lipitor. Therefore, a more practical way to synthesize highly optical active of (S)-CHBE (>99.9% ee) is of great interest. Compared with conventional chemical synthesis, the asymmetric bioreduction of ethyl 4-chloro-3-oxobutanoate (COBE), which is inexpensive and easily synthesized, is an economical approach to the production of (S)-CHBE. From recently research, we find that there is an enzyme can efficiently catalyze this reduction reaction. In addition, this reductase is a NADH-dependent reductase, which perfectly fit our expectation——find an efficient, NADH-dependent reductase as our project can only increase the concentration of NADH inside of the cytoplasm. So, to prove our project’s feasibility, this substrate and product will be a perfect model. We introduce this enzyme to our engineered E.coli to catalyze this reaction.
 +
To induce the gene expression, we use a promotor called pBAD, which is induced by arabinose.
 +
 +
<!-- Add more about the biology of this part here
 +
===Usage and Biology===
 +
 +
<!-- -->
 +
<span class='h3bb'>Sequence and Features</span>
 +
<partinfo>BBa_K2242920 SequenceAndFeatures</partinfo>
 +
 +
 +
<!-- Uncomment this to enable Functional Parameter display
 +
===Functional Parameters===
 +
<partinfo>BBa_K2242920 parameters</partinfo>
 +
<!-- -->

Latest revision as of 02:13, 12 October 2017


araC+pBAD+CmCR

Ethyl (S)-4-chloro-3-hydroxybutanoate [(S)-CHBE] is a key intermediate for the production of chiral drugs, including choles- terol-lowering HMG-CoA reductase inhibitors such as Lipitor. Therefore, a more practical way to synthesize highly optical active of (S)-CHBE (>99.9% ee) is of great interest. Compared with conventional chemical synthesis, the asymmetric bioreduction of ethyl 4-chloro-3-oxobutanoate (COBE), which is inexpensive and easily synthesized, is an economical approach to the production of (S)-CHBE. From recently research, we find that there is an enzyme can efficiently catalyze this reduction reaction. In addition, this reductase is a NADH-dependent reductase, which perfectly fit our expectation——find an efficient, NADH-dependent reductase as our project can only increase the concentration of NADH inside of the cytoplasm. So, to prove our project’s feasibility, this substrate and product will be a perfect model. We introduce this enzyme to our engineered E.coli to catalyze this reaction. To induce the gene expression, we use a promotor called pBAD, which is induced by arabinose.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1205
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 1144
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 1676
    Illegal AgeI site found at 979
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 1935
    Illegal SapI site found at 961