Part:BBa_K4805002
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
- 10COMPATIBLE WITH RFC[10]
- 12INCOMPATIBLE WITH RFC[12]Illegal NheI site found at 676
Illegal NheI site found at 2581 - 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 79
Illegal BamHI site found at 1984 - 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]
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