Difference between revisions of "Part:BBa K2242521"

 
 
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__NOTOC__
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<partinfo>BBa_K2242521 short</partinfo>
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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.
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We use a promoter pTAC to control the gene CmCR’s expression, which is induced by IPTG.
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===Usage and Biology===
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<span class='h3bb'>Sequence and Features</span>
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<partinfo>BBa_K2242521 SequenceAndFeatures</partinfo>
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===Functional Parameters===
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<partinfo>BBa_K2242521 parameters</partinfo>
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Latest revision as of 04:01, 12 October 2017


placI+lacI+pTAC+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. We use a promoter pTAC to control the gene CmCR’s expression, which is induced by IPTG.

Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    COMPATIBLE WITH RFC[12]
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BglII site found at 1353
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    INCOMPATIBLE WITH RFC[25]
    Illegal NgoMIV site found at 3280
  • 1000
    INCOMPATIBLE WITH RFC[1000]
    Illegal BsaI site found at 3539