Ketoreductases (KREDs) Engineering Services

Ketoreductases (KREDs), also known as Alcohol Dehydrogenases (ADHs), are indispensable biocatalysts for the highly selective reduction of prochiral ketones to synthesize Chiral Alcohols. These chiral building blocks are vital intermediates in the production of high-value pharmaceuticals and fine chemicals. While chemical reduction methods often rely on expensive metal catalysts, the utility of native KREDs is often hindered by limited access to specific stereoisomers (R or S), pronounced substrate inhibition at high concentrations, and poor stability in non-aqueous solvent systems necessary for solubility.

Our specialized enzyme engineering services leverage integrated Rational Design and Directed Evolution to overcome these bottlenecks. Our focus areas include: inverting stereoselectivity (e.g., from R-selective to S-selective or vice versa), enhancing stability in industrial organic solvents, and improving activity towards bulky or non-natural ketone substrates. Consult with our experts to design a customized strategy that guarantees the efficient, high-purity synthesis of your target chiral alcohol.

Challenges in Ketoreductase (KRED) Biocatalysis

The successful industrial deployment of KREDs is limited by the following intrinsic factors, which our services are specifically designed to overcome:

• Limited Stereoisomer Access: The vast majority of available KREDs yield only one specific enantiomer (R or S), lacking the flexibility to produce the required alternative stereoisomer.
• Substrate/Product Inhibition: High concentrations of ketone substrates or chiral alcohol products frequently inhibit the enzyme, leading to lower conversion rates and increased cost.
• Low Stability in Organic Systems: For water-insoluble or bulky ketone substrates, non-aqueous or co-solvent systems are required, in which most KREDs exhibit poor stability and activity.
• Activity Towards Non-Natural Substrates: Wild-type KREDs often show low catalytic efficiency (k_cat) towards bulky or synthetically relevant non-natural ketones.

Our engineering platforms focus on addressing these molecular and operational bottlenecks.

Engineering Focus: Stereocontrol and Industrial Robustness

We apply integrated protein engineering strategies to enhance your target Ketoreductase:

Stereoselectivity Inversion Service

             

We engineer the active site to invert the stereopreference, allowing the precise synthesis of the previously inaccessible (R)- or (S)-chiral alcohol with e.e. > 99%.

Solvent Stability Enhancement

Advanced stability engineering services to enhance tolerance and activity in non-aqueous and high-concentration organic co-solvent systems.

Activity and Promiscuity Improvement

Services focused on optimizing catalytic efficiency and substrate range for bulky, challenging, or non-natural ketone substrates.

Inhibition Tolerance Design

We use Rational Design and Directed Evolution to select for variants with reduced sensitivity to substrate and product inhibition at high titers.

Our experts are ready to apply these integrated capabilities to your specific KRED biocatalysis project.

Technology Platforms for KRED Engineering

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

Active Site Engineering for Stereocontrol

Using Active Site Engineering and the Prelog Rule, we modify the large/small substrate pockets to flip the stereoselectivity.

Structural Bioinformatics and Modeling

We use Structural Bioinformatics to model solvent-enzyme interactions and guide the rational design of stability-enhancing mutations.

Integrated Directed Evolution

We apply integrated evolution workflows, utilizing high-throughput screening to identify variants with enhanced performance under challenging conditions (e.g., high organic solvent content).

Comprehensive Enzyme Profiling

We offer full kinetic profiling to accurately determine K_m, V_max, inhibition constants, and solvent/thermal stability under your specific industrial conditions.

Enzyme Immobilization & Formulation

Beyond molecular design, we offer services in stabilization and formulation to ensure the engineered KRED is ready for continuous reactor systems at scale, often involving non-aqueous solvents.

Partner with us to harness these platforms for your next chiral alcohol synthesis breakthrough.

Project Flow: KRED Optimization Workflow

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

• Consultation and Goal Definition: Initial discussion to define precise stereoselectivity (R or S), k_cat, and solvent stability targets for your KRED.
• Design Strategy Proposal: We propose a tailored strategy involving Rational Design (Active Site Engineering) and/or Directed Evolution, outlining the predicted timeframe and resources.
• Library Construction and Screening: We execute mutagenesis (library generation) and employ HTS platforms to identify lead variants that achieve intermediate stereocontrol and stability milestones.
• Iterative Optimization: Successive rounds of evolution are performed under increasingly stringent conditions (e.g., higher organic solvent percentage, high substrate concentration) to build industrial robustness.
• Final Deliverables: Delivery of the final KRED enzyme variant along with detailed stereochemical analysis, performance, and stability 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 KRED requirements and explore how our technologies can achieve their desired enantiopurity targets.

We provide comprehensive support, including:

• Detailed Kinetic Data, Enantiomeric Excess (e.e.), Solvent Tolerance, and Thermal Stability Reports.
• Consultation on process development, including cofactor regeneration system design and integration of the engineered KRED.
• Experimental reports include complete raw data on mutagenesis libraries, screening results, and chiral HPLC traces.