Comprehensive Enzyme Profiling Services

CD Biosynsis offers expert Enzyme Profiling Services essential for the detailed characterization of biocatalysts, therapeutic targets, and novel enzyme variants. Accurate enzyme profiling is the foundational step for both rational engineering and regulatory submissions, providing quantitative data on function, stability, and inhibition. Our integrated platform covers everything from determining fundamental kinetic parameters like Km and Vmax, to mapping the entire substrate scope and rigorously testing inhibitor compounds. We provide detailed stability profiles across various industrial conditions (temperature, pH, solvent), ensuring you have the precise, high-quality data needed to understand your enzyme's full potential, guide optimization efforts, and validate performance metrics.

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Precision Data for Enzyme Function and Validation

Enzyme profiling moves beyond simple activity measurement to provide a full biochemical portrait of a protein. This characterization is crucial for defining the enzyme's mechanism, comparing wild-type performance against engineered variants, and meeting strict requirements for commercial application. We employ state-of-the-art automated assay technology and advanced data analysis software to deliver highly reproducible and accurate results. Whether you are identifying the most promising enzyme from a directed evolution campaign, assessing the off-target effects of a therapeutic enzyme, or determining the shelf-life under specific storage conditions, our services provide the granular detail required for informed decision-making in drug discovery, biocatalysis, and synthetic biology.

Core Enzyme Characterization Modules

Kinetic and Mechanistic Analysis Functional and Substrate Range Stability and Inhibition Assessment

Quantifying Catalytic Efficiency

Kinetic Profiling

Enzyme Kinetic Profiling Service

Precise determination of fundamental Michaelis-Menten parameters (Km, Vmax, kcat) and catalytic efficiency (kcat/Km) using continuous or discontinuous assays and global curve fitting.

Reaction Order and Mechanism

Analysis of multi-substrate enzymes to determine the order of substrate binding and product release (e.g., Ping Pong, Sequential mechanisms) crucial for cascade design.

pH and Temperature Optima

Mapping the optimal pH and temperature conditions for maximal activity to guide process development and storage recommendations.

Defining Substrate Scope and Selectivity

Substrate Profiling

Enzyme Substrate Profiling Service

Testing a panel of natural and non-natural substrates to fully map the enzyme's tolerance, specificity, and selectivity for rational engineering and industrial application.

High-Throughput Substrate Screening

Rapidly screen bespoke libraries of potential substrates using automated liquid handling and customized detection methods.

Stereoselectivity (ee) Analysis

High-precision analysis of enantiomeric excess (ee) via chiral HPLC or GC to validate the chiral purity of enzyme-catalyzed products.

Assessing Robustness and Regulation

Stability and Inhibitor Screening

Enzyme Stability Profiling Service

Determining thermal inactivation half-life (t1/2), Tm (melting temperature), and tolerance to organic solvents, detergents, and high salt concentrations.

Enzyme Inhibitor Screening Service

Screening compound libraries against the enzyme target to determine IC50 and Ki values, defining the inhibition mode (competitive, non-competitive, uncompetitive).

Product and Substrate Inhibition

Characterizing the degree to which accumulated product or high substrate concentrations inhibit enzyme activity, crucial data for reactor design.

Standardized Enzyme Profiling Workflow

Delivering reliable data from sample preparation to detailed reporting.

Enzyme Preparation and Quality Control

Assay Development and Calibration

Kinetic and Substrate Profiling

Data Analysis and Reporting

Sample Receipt: Receive and verify enzyme purity (typically >95 percent by SDS-PAGE or HPLC).

Concentration Determination: Accurately measure active site concentration via titration or Bradford/UV methods.

Assay Selection: Choose the most suitable detection method (spectrophotometric, fluorescent, TRF, or HPLC-based) for the specific reaction.

Standardization: Optimize reaction conditions (buffer, cofactors, pH) to ensure linear reaction rates and minimal background noise.

Execution: Perform multi-point kinetic assays (varying substrate concentrations) and comprehensive stability testing (thermal, pH).

Screening: Run high-throughput screens for substrate panels or inhibitor libraries using automated systems.

Data Fitting: Use non-linear regression software (e.g., GraphPad Prism) for accurate parameter fitting (Km, Vmax, IC50).
Comprehensive Report: Deliver raw data, analyzed curves, mechanistic insights, and detailed methodology protocols.

Precision and Expertise in Enzyme Characterization

High-Fidelity Kinetics

Precise determination of catalytic parameters (Km, kcat) using optimized methods and global fitting software.

Comprehensive Stability Mapping

Detailed profiles including t1/2, Tm, and tolerance to solvents/detergents critical for industrial use.

Advanced Selectivity Analysis

Expert use of chiral HPLC and GC for accurate enantiomeric excess (ee) determination and product purity.

High-Throughput Screening (HTS)

Automated platforms for rapid screening of large substrate or inhibitor libraries (IC50 and Ki determination).

Client Testimonials on Enzyme Profiling Services

"The kinetic profiling was indispensable. We used the precise kcat/Km data to validate our engineered variants, proving a 15-fold improvement in catalytic efficiency."

Dr. Emma Li, Synthetic Biology Lead

"The substrate profiling identified a previously unknown secondary activity, which expanded the commercial application of our enzyme into a new market segment."

Mr. David Cho, Biocatalysis R&D Director

"The detailed stability profiles across various pH and temperature points provided the exact parameters needed for us to confidently transition the enzyme from lab bench to 1000L fermenter."

Ms. Sarah Khan, Process Development Manager

FAQs about Enzyme Profiling Services

Still have questions?

What is the most critical parameter in enzyme kinetic profiling?

The catalytic efficiency (kcat/Km) is generally the most critical parameter as it measures the enzyme's overall performance under low substrate concentration, often reflecting its biological or industrial relevance.

How do you determine the melting temperature (Tm) in stability profiling?

We typically use techniques such as Differential Scanning Fluorimetry (DSF), Circular Dichroism (CD), or Thermal Shift Assays to monitor the unfolding of the enzyme structure as temperature increases.

Can you perform profiling on multi-substrate enzymes?

Yes, we can perform initial velocity studies at varying concentrations of both substrates to determine the kinetic mechanism (e.g., Random Sequential, Ordered Sequential, or Ping Pong) and derive all relevant parameters.

What is the typical output for an inhibitor screening service?

The primary output includes the half maximal inhibitory concentration (IC50), the inhibition constant (Ki), and a determination of the inhibition type (competitive, non-competitive, or uncompetitive).

How much does Metabolic Engineering services cost?

The cost of Metabolic Engineering services depends on the project scope, complexity of the target compound, the host organism chosen, and the required yield optimization. We provide customized quotes after a detailed discussion of your specific research objectives.

Do your engineered strains meet regulatory standards?

We adhere to high quality control standards in all strain construction and optimization processes. While we do not handle final regulatory approval, our detailed documentation and compliance with best laboratory practices ensure your engineered strains are prepared for necessary regulatory filings (e.g., GRAS, FDA).

What to look for when selecting the best gene editing service?

We provide various gene editing services such as CRISPR-sgRNA library generation, stable transformation cell line generation, gene knockout cell line generation, and gene point mutation cell line generation. Users are free to select the type of service that suits their research.

Does gene editing allow customisability?

Yes, we offer very customised gene editing solutions such as AAV vector capsid directed evolution, mRNA vector gene delivery, library creation, promoter evolution and screening, etc.

What is the process for keeping data private and confidential?

We adhere to the data privacy policy completely, and all customer data and experimental data are kept confidential.