Difference between revisions of "Part:BBa K4805002"

 
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__NOTOC__
 
 
<partinfo>BBa_K4805002 short</partinfo>
 
<partinfo>BBa_K4805002 short</partinfo>
 +
==description==
 +
BmeTC is  a tetraprenyl-beta-curcumene Cyclase from Bacillus megaterium (Genbank Accession: CP001982.1, 2130781–2132658). BmeTC_Y167A, D373C refers to a variant of BmeTC that contains two specific mutations: Y167A and D373C. Although BmeTC can only catalyze the bicyclic structure at the end of squalene, its variant BmeTC_D373C and BmeTC_Y167A, D373C possess the ability of catalyzing both monocyclic and bicyclic structure, which can produce ambrein. So far, the synergistic effect of BmeTC_Y167A, D373C and BmeTC in the cell-free system can produce the highest yield of ambrein with squalene as substrate. (Tsutomu S. et al, 2020)
 +
Thus we tried to construct a fusion protein with a flexible linker to tie BmeTC to BmeTC_Y167A, D373C, hoping that this strategy can improve the specifity of the cyclase. Our part can help and inspire other future teams to build and perfect the pathway of producing ambrein from squalene. It belongs to the terpenoids part collection we have established for the production of santalol and ambrein in S. cerevisiae, which includes BBa_K4805000-BBa_K4805012.
  
ATGATAATTTTATTAAAGGAAGTGCAACTTGAAATTCAAAGAAGGATAGCGTACTTAAGACCAACTCAGAAGAATGACGGATCCTTCAGATATTGTTTTGAAACTGGTGTGATGCCAGACGCTTTTCTGATCATGTTGTTACGTACATTCGATTTGGATAAAGAGGTCCTAATCAAGCAATTGACAGAGAGAATTGTCAGCTTACAAAACGAAGATGGTTTATGGACTTTATTTGACGATGAAGAACATAATCTTTCCGCCACTATCCAGGCCTATACGGCTTTGTTATACTCCGGCTATTATCAAAAGAACGACCGTATCTTAAGAAAGGCTGAGAGATACATTATTGATAGTGGTGGGATTTCCCGTGCACATTTCTTGACTAGGTGGATGTTATCTGTGAACGGGCTTTATGAATGGCCAAAATTGTTCTATCTACCTTTATCTTTATTATTAGTGCCTACTTACGTGCCACTTAACTTTTATGAACTGTCTACTTACGCAAGAATCCACTTTGTACCAATGATGGTGGCTGGGAATAAGAAGTTCTCACTGACTTCTCGTCACACTCCATCCTTATCACATCTTGATGTGAGGGAACAAAAGCAAGAAAGTGAAGAAACTACCCAGGAAAGTAGGGCAAGTATTTTTTTGGTCGACCACTTGAAGCAATTGGCTAGCCTGCCCTCATACATACATAAGTTGGGTTACCAAGCTGCTGAAAGATATATGTTGGAAAGGATCGAAAAAGATGGCACTTTGTATTCTTATGCAACGTCCACATTCTTCATGATATACGGCCTTCTTGCTTTAGGTTACAAAAAGGACTCCTTTGTTATACAAAAAGCAATTGATGGCATATGTTCACTGTTATCTACTTGTTCTGGTCATGTTCATGTTGAAAACAGTACTtGTACCGTGTGGGACACCGCTTTGTTATCCTACGCATTACAAGAAGCAGGAGTACCTCAGCAAGATCCTATGATAAAAGGCACGACGAGATACTTAAAAAAAAGACAACACACTAAATTGGGCGATTGGCAATTTCACAATCCTAACACAGCTCCTGGTGGGTGGGGTTTTTCAGATATCAACACTAACAACCCCGACTTAGACGATACCTCAGCTGCTATTAGGGCTTTATCCAGGAGAGCCCAAACTGACACTGACTACTTGGAATCCTGGCAAAGAGGCATTAATTGGCTATTGTCTATGCAAAACAAGGACGGGGGTTTCGCTGCTTTCGAAAAGAATACCGATTCTATACTTTTCACTTATCTGCCGCTAGAGAATGCTAAAGATGCTGCGACAGATCCTGCCACTGCTGACCTAACTGGTAGAGTTCTTGAATGCTTGGGAAATTTTGCAGGCATGAACAAATCACACCCCTCTATAAAAGCTGCCGTCAAGTGGTTGTTTGATCACCAACTTGACAACGGTAGTTGGTACGGCAGATGGGGTGTATGCTACATTTATGGTACATGGGCTGCCATAACAGGCTTAAGAGCTGTAGGCGTGTCTGCTTCTGATCCCAGAATTATTAAGGCTATCAATTGGCTAAAGTCAATCCAACAAGAAGATGGTGGGTTCGGGGAATCTTGTTATTCTGCCTCTTTGAAGAAATATGTGCCGCTGAGTTTTAGTACGCCCTCTCAGACAGCCTGGGCTTTAGACGCCCTGATGACAATATGTCCATTGAAGGACCGTTCAGTGGAAAAAGGTATTAAGTTCCTGTTAAACCCGAATTTGACAGAGCAGCAGACCCATTATCCAACGGGTATAGGTTTACCGGGACAATTTTATATACAGTATCACAGCTATAATGACATTTTTCCTTTGCTTGCTCTGGCTCATTACGCCAAAAAGCATTCCAGTggtggcggaggttcgggtggcggtgGATCTATGATAATTTTATTAAAGGAAGTGCAACTTGAAATTCAAAGAAGGATAGCGTACTTAAGACCAACTCAGAAGAATGACGGATCCTTCAGATATTGTTTTGAAACTGGTGTGATGCCAGACGCTTTTCTGATCATGTTGTTACGTACATTCGATTTGGATAAAGAGGTCCTAATCAAGCAATTGACAGAGAGAATTGTCAGCTTACAAAACGAAGATGGTTTATGGACTTTATTTGACGATGAAGAACATAATCTTTCCGCCACTATCCAGGCCTATACGGCTTTGTTATACTCCGGCTATTATCAAAAGAACGACCGTATCTTAAGAAAGGCTGAGAGATACATTATTGATAGTGGTGGGATTTCCCGTGCACATTTCTTGACTAGGTGGATGTTATCTGTGAACGGGCTTTATGAATGGCCAAAATTGTTCTATCTACCTTTATCTTTATTATTAGTGCCTACTTACGTGCCACTTAACTTTTATGAACTGTCTACTgctGCAAGAATCCACTTTGTACCAATGATGGTGGCTGGGAATAAGAAGTTCTCACTGACTTCTCGTCACACTCCATCCTTATCACATCTTGATGTGAGGGAACAAAAGCAAGAAAGTGAAGAAACTACCCAGGAAAGTAGGGCAAGTATTTTTTTGGTCGACCACTTGAAGCAATTGGCTAGCCTGCCCTCATACATACATAAGTTGGGTTACCAAGCTGCTGAAAGATATATGTTGGAAAGGATCGAAAAAGATGGCACTTTGTATTCTTATGCAACGTCCACATTCTTCATGATATACGGCCTTCTTGCTTTAGGTTACAAAAAGGACTCCTTTGTTATACAAAAAGCAATTGATGGCATATGTTCACTGTTATCTACTTGTTCTGGTCATGTTCATGTTGAAAACAGTACTtGTACCGTGTGGGACACCGCTTTGTTATCCTACGCATTACAAGAAGCAGGAGTACCTCAGCAAGATCCTATGATAAAAGGCACGACGAGATACTTAAAAAAAAGACAACACACTAAATTGGGCGATTGGCAATTTCACAATCCTAACACAGCTCCTGGTGGGTGGGGTTTTTCAGATATCAACACTAACAACCCCGACTTAGACtgTACCTCAGCTGCTATTAGGGCTTTATCCAGGAGAGCCCAAACTGACACTGACTACTTGGAATCCTGGCAAAGAGGCATTAATTGGCTATTGTCTATGCAAAACAAGGACGGGGGTTTCGCTGCTTTCGAAAAGAATACCGATTCTATACTTTTCACTTATCTGCCGCTAGAGAATGCTAAAGATGCTGCGACAGATCCTGCCACTGCTGACCTAACTGGTAGAGTTCTTGAATGCTTGGGAAATTTTGCAGGCATGAACAAATCACACCCCTCTATAAAAGCTGCCGTCAAGTGGTTGTTTGATCACCAACTTGACAACGGTAGTTGGTACGGCAGATGGGGTGTATGCTACATTTATGGTACATGGGCTGCCATAACAGGCTTAAGAGCTGTAGGCGTGTCTGCTTCTGATCCCAGAATTATTAAGGCTATCAATTGGCTAAAGTCAATCCAACAAGAAGATGGTGGGTTCGGGGAATCTTGTTATTCTGCCTCTTTGAAGAAATATGTGCCGCTGAGTTTTAGTACGCCCTCTCAGACAGCCTGGGCTTTAGACGCCCTGATGACAATATGTCCATTGAAGGACCGTTCAGTGGAAAAAGGTATTAAGTTCCTGTTAAACCCGAATTTGACAGAGCAGCAGACCCATTATCCAACGGGTATAGGTTTACCGGGACAATTTTATATACAGTATCACAGCTATAATGACATTTTTCCTTTGCTTGCTCTGGCTCATTACGCCAAAAAGCATTCCAGTTGA
 
  
  

Revision as of 15:08, 12 October 2023

BmeTC-FL-BmeTC Y167A,D373C

description

BmeTC is a tetraprenyl-beta-curcumene Cyclase from Bacillus megaterium (Genbank Accession: CP001982.1, 2130781–2132658). BmeTC_Y167A, D373C refers to a variant of BmeTC that contains two specific mutations: Y167A and D373C. Although BmeTC can only catalyze the bicyclic structure at the end of squalene, its variant BmeTC_D373C and BmeTC_Y167A, D373C possess the ability of catalyzing both monocyclic and bicyclic structure, which can produce ambrein. So far, the synergistic effect of BmeTC_Y167A, D373C and BmeTC in the cell-free system can produce the highest yield of ambrein with squalene as substrate. (Tsutomu S. et al, 2020) Thus we tried to construct a fusion protein with a flexible linker to tie BmeTC to BmeTC_Y167A, D373C, hoping that this strategy can improve the specifity of the cyclase. Our part can help and inspire other future teams to build and perfect the pathway of producing ambrein from squalene. It belongs to the terpenoids part collection we have established for the production of santalol and ambrein in S. cerevisiae, which includes BBa_K4805000-BBa_K4805012.


Sequence and Features


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 676
    Illegal NheI site found at 2581
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 79
    Illegal BamHI site found at 1984
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]