Difference between revisions of "Part:BBa K4805009:Design"

 
 
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<partinfo>BBa_K4805009 SequenceAndFeatures</partinfo>
 
<partinfo>BBa_K4805009 SequenceAndFeatures</partinfo>
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==Description==
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BmeTC_Y167A, D373C refers to a variant of the Tetraprenyl-beta-curcumene Cyclase from Bacillus megaterium (Genbank Accession: CP001982.1, 2130781–2132658) 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, it has been used in cell-free system with squalene as substrate, and can produce the highest yield of ambrein in all single enzyme reactions. (Tsutomu S. et al, 2020)
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Our part can help and inspire other future teams to build and perfect the pathway of producing ambrein from squalene in yeast, with the regulation of pTDH3 and tTDH1. It belongs to the part collection we have established for the production of santalol and ambrein in S. cerevisiae, which includes BBa_K4805000-BBa_K4805012.
  
  
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DNA synthesis
 
DNA synthesis
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===References===
 

Latest revision as of 15:24, 12 October 2023


pTDH3-BmeTC-FL-BmeTC_Y167A,D373C-tTDH1


Assembly Compatibility:
  • 10
    COMPATIBLE WITH RFC[10]
  • 12
    INCOMPATIBLE WITH RFC[12]
    Illegal NheI site found at 1357
    Illegal NheI site found at 3262
  • 21
    INCOMPATIBLE WITH RFC[21]
    Illegal BamHI site found at 760
    Illegal BamHI site found at 2665
    Illegal XhoI site found at 4472
  • 23
    COMPATIBLE WITH RFC[23]
  • 25
    COMPATIBLE WITH RFC[25]
  • 1000
    COMPATIBLE WITH RFC[1000]

Description

BmeTC_Y167A, D373C refers to a variant of the Tetraprenyl-beta-curcumene Cyclase from Bacillus megaterium (Genbank Accession: CP001982.1, 2130781–2132658) 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, it has been used in cell-free system with squalene as substrate, and can produce the highest yield of ambrein in all single enzyme reactions. (Tsutomu S. et al, 2020)

Our part can help and inspire other future teams to build and perfect the pathway of producing ambrein from squalene in yeast, with the regulation of pTDH3 and tTDH1. It belongs to the part collection we have established for the production of santalol and ambrein in S. cerevisiae, which includes BBa_K4805000-BBa_K4805012.


Design Notes

Its DNA sequences have been optimized which can be expressed in S. cerevisiae.


Source

DNA synthesis