Parts Collection
Our project focuses on adopting various strategies to enhance the efficiency of squalene synthesis by Yarrowia Lipolytica using fatty acids as substrates. The project aims to utilize low-cost and abundant WO (Waste oil) for squalene synthesis, thereby meeting the growing demand of the cosmetics industry for squalene, an excellent raw material. To address the feedback inhibition and interference affecting the MVA pathway in the cytoplasm, we have proposed a compartmentalization strategy.
To facilitate the application of this compartmentalization strategy in the actual production of squalene and the potential synthesis of other terpenoids in the future, we have developed a parts collection specifically for peroxisomal compartmentalization. (UUID:6a1f3d92-50e1-4192-bd4f-e652a0293224)This collection includes:
Overexpression parts for upregulating the beta-oxidation pathway that inherently exists in peroxisomes (BBa_25VW58NU );
A set of multi-gene integration parts for compartmentalizing the MVA pathway via the PTS1 sequence(BBa_25T8A2CI BBa_25KXC0C8 BBa_25P1VJS7);
Dynamic regulatory parts that increase the size and quantity of peroxisomes(BBa_25OGDY94).
By applying this series of parts, the compartmentalization process of the MVA pathway becomes simple and efficient. Against the backdrop of the increasing widespread application of squalene—as an antioxidant and moisturizing raw material—this compartmentalization scheme holds great promise. Meanwhile, it can serve as a cornerstone for almost all MVA pathway-related synthesis projects and is expected to become a key component of future solutions for terpenoid synthesis.
In the context where the cosmetics industry is placing greater emphasis on environmental protection and sustainability, the parts collection developed in our project addresses the issue that environmental requirements have restricted the widespread application of squalene. It also aims to reduce the cost of squalene and provides a feasible strategy for the biosynthetic production of other raw materials in the fashion industry.
We encourage and welcome other teams to use our parts collection for the compartmentalized synthesis of any products. Building on the foundation we have established, they can further characterize the enzyme properties in the pathway to optimize the compartment environment for improved reaction efficiency, or optimize the energy supply in peroxisomes and establish dynamic regulatory mechanisms. Such efforts will help refine this compartmentalization strategy.
Basic Parts
| Part Name | Registry Code | Part Type |
|---|---|---|
| NADH-specific HMGr | BBa_25ID9C9T | Coding |
| hp4d promoter | BBa_25Z23TWP | Coding |
| Truncated HMGr | BBa_25QI6SNO | Coding |
| mTLL | BBa_25PDYT4M | Coding |
| ERG20 farnesyl-pyrophosphate synthetase | BBa_25PKX45N | Coding |
| PEX10 Peroxisomal biogenesis factor 10 (Peroxin-10) | BBa_25QTETEL | Coding |
| crRNA(pex23) | BBa_256CNBY9 | gRNA_gene [SO:0001264] |
| crRNA(ERG1) | BBa_2527XAFW | gRNA_gene [SO:0001264] |
| crRNA(Hsp82 and Hsc 82) | BBa_255SF300 | gRNA_gene [SO:0001264] |
| EL222 | BBa_256XOZ39 | Coding |
| PhyB | BBa_2554ZTPY | Coding |
| PIF-AD(PIF3) | BBa_25XNB1XO | Coding |
| LexA | BBa_25GBW6WP | Coding |
| PIF-BP-minimal box promoter | BBa_25JBULLH | Promoter |
| PTS1 (codons optimized for Yarrowia lipolytica) | BBa_257EN8OW | Signalling |
| 5C120-8lexA operator-CYC1 minimal box promoter | BBa_25A851G4 | Promoter |
| IDI1 isopentenyl-diphosphate delta-isomerase | BBa_256KAMKD | Coding |
| ERG9 SQS1 Squalene synthase | BBa_25IB8LCD | Coding |
| ERG19 MVD1 mevalonate pyrophosphate decarboxylase | BBa_25ZJWX87 | Coding |
| ERG8 phosphomevalonate kinase | BBa_25Y0DD6A | Coding |
| ERG12 mevalonate kinase singleton | BBa_25FSSLG6 | Coding |
| ERG13 3-hydroxy-3-methylglutaryl-CoA (HMG-CoA) synthase | BBa_25T4M46S | Coding |
| ERG10 Acetyl-CoA C-acetyltransferase (acetoacetyl-CoA thiolase) | BBa_25HF9GLR | Coding |
| POT1 3-ketoacyl-CoA thiolase with broad chain length specificity | BBa_25K8LHYY | Coding |
| POX2 Fatty-acyl coenzyme A oxidase | BBa_25UJCQ2N | Coding |
| XPR2_term | BBa_25KOAW9U | Terminator [SO:0000141] |
| Kozak | BBa_K5073037 | Scar |
| ylMFE1 | BBa_K3909006 | Coding |
Composite Parts
| Part Name | Registry Code | Part Type |
|---|---|---|
| Hp4d-Kozak-IDI-PTS1-XPR2_term | BBa_25D2R2WL | Plasmid |
| Hp4d-Kozak-ERG9-PTS1-XPR2_term | BBa_25SV6BI3 | Plasmid |
| Hp4d-Kozak-ERG12-PTS1-XPR2_term | BBa_25HRKCCB | Plasmid |
| Hp4d-Kozak-ERG13-PTS1-XPR2_term | BBa_25UYC74A | Plasmid |
| Hp4d-Kozak-ERG10-PTS1-XPR2_term | BBa_25R8GVCZ | Plasmid |
| Hp4d-Kozak-ERG19-PTS1-XPR2_term | BBa_25AWQFNS | Plasmid |
| Hp4d-Kozak-tHMGr-XPR2_term | BBa_258W49OO | Plasmid |
| Hp4d-Kozak-NADH_HMGr -XPR2_term | BBa_256WCABY | Plasmid |
| HP4d-Kozak-PEX10-XPR2-term | BBa_25OGDY94 | Plasmid |
| Hp4d-Kozak-POX2-XPR2_term | BBa_25NF8WSH | Plasmid |
| Hp4d-Kozak-MFE2-XPR2_term | BBa_25P40PG4 | Plasmid |
| Hp4d-Kozak-POT1-XPR2_term | BBa_256UU7WP | Plasmid |
| Hp4d-Kozak-tHMGr-PTS1-XPR2_term | BBa_257C5P3N | Plasmid |
| Hp4d-Kozak-NADH_HMGr -PTS1-XPR2_term | BBa_25SUXQ01 | Plasmid |
| Hp4d-Kozak-ERG8-PTS1-XPR2_term | BBa_256XVAWC | Plasmid |
| Hp4d-Kozak-ERG20-PTS1-XPR2_term | BBa_25RNKI3F | Plasmid |
| Hp4d-Kozak-mTLL-XPR2_term | BBa_25UL4UCP | Plasmid |
| Hp4d-Kozak-POX2-XPR2_term-Hp4d-Kozak-MFE1-XPR2_term-Hp4d-Kozak-POT1-XPR2_term | BBa_25VW58NU | Plasmid |
| Hp4d-Kozak-ERG10-PTS1-XPR2_term-Hp4d-Kozak-ERG13-PTS1-XPR2_term-Hp4d-Kozak-tHMGR-PTS1-XPR2_term-Hp4d | BBa_25T8A2CI | Plasmid |
| Hp4d-Kozak-ERG12-PTS1-XPR2_term-Hp4d-Kozak-ERG8-PTS1-XPR2_term-Hp4d-Kozak-ERG19-PTS1-XPR2_term | BBa_25P1VJS7 | Plasmid |
| Hp4d-Kozak-E9-PTS1-XPR2_term-Hp4d-Kozak-IDI-PTS1-XPR2_term-Hp4d-Kozak-E20-PTS1-XPR2_term | BBa_25KXC0C8 | Plasmid |
| Modular Light-Controlled CRISPR | BBa_25A20MN1 | Plasmid |
| HP4d-PhyB- XPR2_term -hp4d-PIF-XPR2_term-PIF_BP-LexA- XPR2_term | BBa_25DPZ6QD | Plasmid |