Nitrilases / Nitrile Hydratases Engineering Services

Nitrilases and Nitrile Hydratases (NHases) are crucial biocatalysts for the green synthesis of chiral carboxylic acids and amides, key intermediates in the food, agricultural, and pharmaceutical sectors, notably in Chiral Acid/Amide Synthesis (e.g., L-Aspartic Acid). Addressing the challenges of traditional methods—such as low stereoselectivity, product inhibition, and environmental sensitivity—requires advanced protein modification techniques.

CD Biosynsis offers specialized enzyme engineering services and powerful technology platforms to tackle these limitations for your specific Nitrilase or NHase project. Our capabilities are focused on achieving Rational Design for enhanced enantioselectivity (> 99% ee); Increased stability in harsh industrial environments; Improved catalytic rate and Km value. We invite you to consult with our experts to design a customized strategy that transforms your enzyme's performance for your unique industrial needs.

Challenges in Nitrilase/NHase Development

The successful industrial deployment of Nitrilases/NHases is limited by these key factors, which our services are designed to overcome:

• Low Stereoselectivity: Many enzymes can yield racemic mixtures, lacking the specificity required for high-purity chiral intermediates.
• Product Inhibition: High concentrations of acid or amide products frequently inhibit the enzyme, forcing lower titer and higher cost.
• Environmental Sensitivity: Enzymes often show poor stability against industrial stresses like non-optimal pH and temperature variations.
• Suboptimal Catalytic Rate: For cost-effective scaling, the turnover rate kcat and substrate affinity Km must be optimized far beyond wild-type levels.

Our engineering platforms focus on addressing these molecular bottlenecks.

Engineering Focus

CD Biosynsis applies integrated protein engineering strategies to enhance your target Nitrilase/NHase:

Stereoselectivity Refinement Service

           

We offer services to engineer the enzyme's chiral selectivity, aiming for enantiomeric excess e.e. > 99\% by precisely modifying the active site.

Stability and Tolerance Enhancement Service

We provide advanced engineering services to increase the enzyme stability against high titer, pH excursions, and thermal stress.

Kinetic Parameter Optimization

Our services focus on optimizing kcat and Km values through site-specific mutagenesis, targeting higher turnover and substrate affinity.

Rational and Directed Approaches

We integrate Rational Design and Directed Evolution workflows to explore the mutational landscape for optimal variants rapidly and systematically.

Our experts are ready to apply these capabilities to your specific enzyme candidate.

Technology Platforms

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

High-Throughput Screening

Platforms like Fluorescence-Activated Droplet Sorting (FADS) and microtiter plate screening enable rapid evaluation of millions of enzyme variants for desired activity and selectivity.

Computational Rational Design

Using Computer-Aided Rational Design (CARD) services, we can predict and prioritize key mutations that maximize substrate fit and chiral bias before lab work begins at the active site or allosteric sites.

Automated Evolution Technologies

Platforms such as Phage-Assisted Continuous Evolution (PACE) accelerate the evolutionary process to generate optimized variants in a fraction of the traditional time.

Comprehensive Kinetic Profiling

We offer full enzyme profiling services to accurately determine kinetic parameters Km, Vmax and operational stability under your specific industrial conditions.

Enzyme Immobilization and Formulation

Beyond molecular design, we offer services in stabilization and immobilization to ensure the engineered enzyme is ready for continuous flow or reusable reactor systems at scale up stage of production process.

Partner with us to harness these platforms for your next biocatalysis breakthrough.

Project Flow

Our enzyme optimization projects follow a flexible, milestone-driven workflow customized to your target goals:

• Consultation and Goal Definition: Initial discussion with our scientific team to define precise kcat, Km, and e.e. targets for your Nitrilase/NHase.
• Design Strategy Proposal: We propose a tailored strategy involving CARD or Directed Evolution, outlining the predicted timeframe and resources.
• Library Construction and Screening: We execute the mutagenesis and employ our HTS platforms to identify lead variants that meet intermediate milestones.
• Iterative Optimization: Successive rounds of evolution are performed under increasingly stringent conditions e.g. high product or suboptimal pH to build robustness.
• Final Deliverables: Delivery of the final enzyme variant along with detailed characterization and performance reports ready for your scale-up process.

Technical communication is maintained throughout the project. We encourage potential clients to initiate a consultation to discuss their specific Nitrilase/NHase requirements and explore how our technologies can achieve their desired yield and purity targets.

CD Biosynsis provides comprehensive support, including:

• Detailed Kinetic Data, Enantiomeric Excess e.e., Product Tolerance, and Stability Reports.
• Consultation on process development and integration of the engineered enzyme into existing chemical or fermentation routes.
• Experimental reports include complete raw data on mutagenesis libraries, screening results, and chiral HPLC traces.