Alkyl Synthases/Alkane Monooxygenases Engineering Services

Alkyl Synthases and Alkane Monooxygenases (e.g., in P450 systems) are key enzymatic players in the sustainable production of fuels, particularly in the synthesis of Bio-jet Fuel and other valuable Alkane Production pathways. These biocatalysts offer a clean route to medium- to long-chain hydrocarbons. However, their industrial viability is severely limited by low chain-length specificity (resulting in mixed products), poor yield of the desired alkane/alcohol length (e.g., C10-C16), and the high cost/inefficiency of co-factor regeneration (e.g., NADPH for P450s).

Our specialized enzyme engineering services are dedicated to resolving these selectivity and cost challenges. Our core objectives include: Rational Redesign of the active site to dictate specific chain termination (improving C10-C16 purity), optimizing P450 systems for highly efficient NADPH use, and improving catalytic activity towards long-chain fatty acid precursors. Consult with our experts to design a customized strategy that elevates the specificity and efficiency of your alkane biosynthesis pathway.

Challenges in Alkane Biosynthesis Biocatalysis

The transition of Alkyl Synthases and Monooxygenases to commercial scale is hindered by these key issues:

• Low Chain-Length Specificity: Enzymes often produce a mixture of short, medium, and long-chain alkanes/alcohols, dramatically complicating downstream purification and lowering the yield of the target fuel fraction (e.g., C10-C16 jet fuel).
• Low Yields: The overall catalytic efficiency (turnover) towards long-chain fatty acid precursors is often suboptimal, resulting in low final titers in microbial hosts.
• Co-factor Cost and Regeneration: Monooxygenases, such as P450 systems, require expensive co-factors like NADPH, making efficient regeneration and utilization essential for economic viability.
• Active Site Substrate Access: Achieving efficient access and processing of hydrophobic long-chain substrates within the active site tunnel can be challenging.

Our engineering platforms are dedicated to resolving these critical specificity and efficiency limitations.

Engineering Focus: Chain-Length Precision and Co-factor Efficiency

We apply integrated protein engineering strategies to enhance your target Alkyl Synthase or Monooxygenase:

Targeted Chain-Length Specificity

             

We use active site engineering to redesign the hydrophobic tunnel, enforcing specific chain termination for C10-C16 production.

Catalytic Activity and Yield Improvement

Advanced optimization services to improve the k_cat towards long-chain fatty acids, thereby increasing the final alkane yield.

Co-factor Optimization (NADPH)

We optimize P450 systems through cofactor engineering and fusion proteins to maximize the efficiency of expensive NADPH utilization.

Solvent Tolerance and Stability

Engineering for high stability and activity in the presence of the hydrocarbon product and any necessary organic solvents for recovery, using stability engineering.

Our experts are ready to apply these integrated capabilities to your specific bio-jet fuel or alkane production pathway.

Technology Platforms for Alkane Enzyme Engineering

We leverage a suite of cutting-edge platforms to deliver highly functional enzyme variants:

Computational Rational Design (CARD)

Using structural modeling and simulation, we precisely redesign the active site tunnel size to dictate the terminal length of the hydrocarbon product.

Directed Evolution for Selectivity

We implement HTS screening using GC-MS or other analytical methods to rapidly select variants with narrow product chain-length distribution.

High-Throughput Enzyme Discovery

We leverage computational mining and functional screening to identify novel Alkyl Synthases with natural preference for C10-C16 substrates.

Comprehensive Enzyme Profiling

We offer full kinetic profiling, including detailed product distribution analysis and efficiency of fatty acid substrate conversion.

Integrated Enzyme Production

Specialized custom production services to achieve high yield and purity for membrane-associated Monooxygenases (P450s) in suitable host systems.

Partner with us to harness these platforms for your bio-jet fuel innovation.

Project Flow: Alkyl Synthase Optimization Workflow

Our enzyme optimization projects follow a flexible, milestone-driven workflow:

• Consultation and Goal Definition: Initial discussion to define the precise chain-length target (e.g., > 90% C14-C16) and required co-factor utilization efficiency.
• Design Strategy Proposal: We propose a tailored strategy involving Rational Design (focused on the catalytic pocket geometry) and/or Directed Evolution under selective pressure, outlining the predicted timeframe.
• Library Construction and Screening: We execute targeted mutagenesis and employ HTS coupled with sensitive analytical techniques to identify lead variants with enhanced chain-length specificity.
• Iterative Optimization & Profiling: Successive rounds of evolution focus on maximizing activity towards long-chain fatty acids and optimizing enzyme expression within the target microbial host.
• Final Deliverables: Delivery of the final enzyme variant along with detailed product distribution, kinetic, and co-factor utilization reports ready for pathway integration.

Technical communication is maintained throughout the project. We encourage potential clients to initiate a consultation to discuss their specific hydrocarbon production requirements and explore how our technologies can achieve high purity and yield.

We provide comprehensive support, including:

• Detailed Kinetic Data, Product Distribution Analysis (e.g., GC/MS), and Co-factor Utilization Efficiency Reports.
• Consultation on metabolic pathway integration and optimization of upstream fatty acid supply.
• Experimental reports include complete raw data on mutagenesis libraries, screening results, and product analysis traces.