Difference between revisions of "Part:BBa K4274033"
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==Usage and biology== | ==Usage and biology== | ||
| − | <i>Cl</i>SS_S533A (C. lansium santalene synthase) is the enzyme that catalyzes the transformation from FPP (farnesyl diphosphate) to alpha-santalene. Its 533<sup>rd amino acid has been artificially mutated from serine to alanine for the sake of optimization. ERG20_F96W is the enzyme that catalyzes the formation of FPP (farnesyl diphosphate). The 86<sup>th amino acid was mutated from phenylalanine to tryptophan. FL refers to the flexible linker that we have added in between the two proteins. It was hoped that adding a flexible linker can maximize alpha-santalene production. However, our results demonstrate the opposite, that alpha-santalene production decreased after adding a flexible linker. | + | <i>Cl</i>SS_S533A (C. lansium santalene synthase) is the enzyme that catalyzes the transformation from FPP (farnesyl diphosphate) to alpha-santalene. Its 533<sup>rd</sup> amino acid has been artificially mutated from serine to alanine for the sake of optimization. ERG20_F96W is the enzyme that catalyzes the formation of FPP (farnesyl diphosphate). The 86<sup>th/sup> amino acid was mutated from phenylalanine to tryptophan. FL refers to the flexible linker that we have added in between the two proteins. It was hoped that adding a flexible linker can maximize alpha-santalene production. However, our results demonstrate the opposite, that alpha-santalene production decreased after adding a flexible linker. |
==Source== | ==Source== | ||
Revision as of 11:36, 12 October 2022
ptac-RiboJ-B0034-ClSS_S533A-FL-ERG20_F96W-B0015
BBa_K3552015, BBa_B0034, BBa_K4274003,BBa_B0015
Usage and Biology
This composite part is designed for the pathway IPP ⇄DMAPP → GPP → FPP → alpha-santalene. It is used as a trial to express enzymes ClSS and ERG20 to its maximized extent, hence reaching the improvement of alpha-santalene production. It starts with the promoter ptac-RiboJ (Part: BBa_K3552015), following the ribosome binding site B0034 (Part: B0034). Then, we added the coding sequence for ClSS_S533A, followed by a flexible linker, and then ERG20_F96W. The part ends with the terminator B0015 (Part: BBa_B0015).
This part falls under our collection that produces alpha-santalene. We also have two parts that optimizes the activity of FPP (farnesyl diphosphate), consisting of ERG20_F96W (Part: BBa_K4274002), and ptac-RiboJ-B0034-ClSS_S533A-B0034-ERG20-B0015 (Part: BBa_K4274030). Parts secreting ClSS and ERG20, CmR29M2_ClSS_S533A (PART: BBa_K4274001) and ptac-RiboJ-B0034-CmR29M2_ClSS_S533A-B0034-ERG20_F96W-B0015 (Part: BBa_K4274032). Finally, this part, exploring the effects of fusion protein on secretion and santalene production (Part: BBa_K4274033).
This part collection aims to optimize the production of alpha-santalene, the precursor of santalol.
Usage and biology
ClSS_S533A (C. lansium santalene synthase) is the enzyme that catalyzes the transformation from FPP (farnesyl diphosphate) to alpha-santalene. Its 533rd amino acid has been artificially mutated from serine to alanine for the sake of optimization. ERG20_F96W is the enzyme that catalyzes the formation of FPP (farnesyl diphosphate). The 86th/sup> amino acid was mutated from phenylalanine to tryptophan. FL refers to the flexible linker that we have added in between the two proteins. It was hoped that adding a flexible linker can maximize alpha-santalene production. However, our results demonstrate the opposite, that alpha-santalene production decreased after adding a flexible linker.
Source
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BglII site found at 3943
Illegal BglII site found at 4004 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 1541
Illegal AgeI site found at 3659 - 1000COMPATIBLE WITH RFC[1000]