Isoprenoids/Terpenoids Bioproduction Engineering Service

Isoprenoids (or Terpenoids) are the largest class of natural products, serving as vital ingredients in cosmetics , perfumes (e.g., Limonene, Farnesene), and pharmaceuticals (e.g., Artemisinin). Industrial sourcing faces two key constraints. Firstly, Natural plant extracts are inconsistent, low-purity , and the yield is highly dependent on climate , leading to supply chain instability. Secondly, conventional petrochemical synthesis involves multi-step processes with toxic reagents, leading to high cost fluctuations and environmental concerns . The key challenge for microbial production is ensuring sufficient supply of the universal Isoprenoid precursors, Isopentenyl Pyrophosphate (IPP) and Dimethylallyl Pyrophosphate (DMAPP) .

CD Biosynsis offers a comprehensive metabolic engineering solution, typically utilizing engineered E. coli (MEP pathway) or yeast (MVA pathway) as hosts. To overcome the precursor bottleneck, we implement Overexpress rate-limiting enzymes in the host's Mevalonate (MVA) or Methylerythritol Phosphate (MEP) pathways to dramatically increase the IPP /text{DMAPP precursor pool . For E. coli, this involves overexpressing DXS and IspF; for yeast, this involves HMG-CoA reductase. To direct this massive precursor flux to the desired Terpenoid product, we Introduce the target-specific Terpene Synthase (e.g., Farnesene Synthase) to channel the high precursor flux to the desired final product . This targeted engineering ensures the production of high-titer, pure Terpenoids independent of volatile natural or petrochemical sources.

Pain Points

Industrial Isoprenoid production faces these key challenges:

• Inconsistency and Low Purity from Natural Extracts: Plant extraction yields are highly variable due to climate and season, and the extract often contains a complex mixture of isomers and impurities , complicating downstream separation.
• High Cost Fluctuations and Environmental Concerns: Petrochemical synthesis routes are tied to oil prices, leading to high cost fluctuations . The processes involve toxic reagents and are generally environmentally unfriendly.
• Limiting Precursor Supply: The initial flux from primary metabolism to the universal precursors IPP and DMAPP is naturally low in microbial hosts, which limits the activity of the downstream Terpene Synthase, leading to low Terpenoid titer.
• Pathway Competition: IPP and DMAPP are essential for native cellular function (e.g., cell membrane sterols in yeast), leading to competitive consumption of the precursors and reduced target product yield.

A successful solution must ensure a stable, massive supply of the IPP /text{DMAPP precursor and direct it to the target product.

Solutions

CD Biosynsis utilizes advanced metabolic engineering to optimize Isoprenoid production:

Overexpress Rate-Limiting Enzymes (DXS, HMG-CoA Reductase)

           

We introduce highly active MVA or MEP genes and overexpress rate-limiting steps (e.g., HMG-CoA reductase in yeast), boosting the IPP /text{DMAPP pool size.

Introduce Target-Specific Terpene Synthase (e.g., Farnesene Synthase)

We select and optimize a Terpene Synthase (TS) gene to convert the high IPP /text{DMAPP flux into the single, desired final Terpenoid product.

Block Competitive Native Pathways

We use gene knockouts (e.g., ERG9 in yeast for Sterol synthesis) to prevent precursor diversion into non-Terpenoid products.

Subcellular Compartmentalization and Product Efflux

We target enzymes to specific organelles (e.g., peroxisome) and introduce efflux pumps to increase yield and reduce product toxicity to the host.

This systematic approach overcomes precursor limitations and precisely channels flux to the target Isoprenoid.

Advantages

Our Isoprenoid engineering service is dedicated to pursuing the following production goals:

High and Stable Yield

MVA /text{MEP pathway boosting ensures a massive and stable IPP /text{DMAPP supply, overcoming the low yield inconsistency of natural extraction.

High Purity and Single Isomer Icon

The highly selective Terpene Synthase produces primarily a single isomer (e.g., trans-Farnesene), simplifying purification compared to plant extracts.

Environmentally Friendly Production Icon

Microbial fermentation replaces chemical synthesis, eliminating toxic reagents and reducing dependency on petrochemicals.

Flexible Product Targeting Icon

By simply switching the Terpene Synthase gene, the same engineered precursor host can produce a wide range of Terpenoids (Mono-, Sesqui-, Diterpenes).

Cost Control and Stability Icon

Microbial production uses low-cost glucose feedstock, achieving stable cost control independent of volatile oil and plant markets.

We provide a sustainable, high-purity, and cost-stable platform for Isoprenoid manufacturing.

Process

Our Isoprenoid strain engineering service follows a rigorous, multi-stage research workflow:

• Precursor Pathway Engineering: Introduce and overexpress key genes in the MVA or MEP pathway (e.g., HMG1 in yeast, dxs in E. coli) to achieve high IPP /text{DMAPP titer.
• Terminal Enzyme Introduction: Integrate the target-specific Terpene Synthase gene (e.g., Limonene Synthase or Farnesene Synthase) and optimize its expression level.
• Native Pathway Blockade: Knockout genes responsible for competing endogenous products (e.g., ERG9 for Sterols in yeast) to maximize carbon flux to the target.
• Product Efflux Optimization: Engineer the host membrane and lipid droplets to facilitate the non-toxic export or storage of the final hydrophobic Terpenoid product.
• Titer and Purity Validation: Validate the engineered strain in fed-batch culture, measuring the final Isoprenoid titer (g/L) and the isomer purity (GC-MS).

Technical communication is maintained throughout the process, focusing on timely feedback regarding yield and product quality attributes.

Explore the potential for a high-titer, high-purity Isoprenoid supply. CD Biosynsis provides customized strain and process engineering solutions:

• Detailed IPP /text{DMAPP Intracellular Concentration Report , demonstrating the effect of pathway boosting.
• Consultation on optimized fermentation strategies (e.g., two-phase fermentation with organic solvent overlay) for volatile Terpenoid products.
• Experimental reports include complete raw data on Terpenoid titer, yield, and final isomer purity , crucial for fragrance and supplement applications.