Enzyme Activity Measurement for Hydrolases

inquiry

Hydrolases are one of the most widely studied and industrially applied classes of enzymes, catalyzing the cleavage of chemical bonds through hydrolysis. They play essential roles in metabolism, digestion, and biocatalysis. At Creative Enzymes, we provide Enzyme Activity Measurement Services for Hydrolases to deliver precise, reproducible, and insightful data that supports both academic research and industrial applications.

Understanding Hydrolases Activity Measurement

Hydrolases catalyze the hydrolytic cleavage of bonds such as ester, glycosidic, peptide, and other covalent linkages. Their broad functionality makes them indispensable in diverse areas, including:

Food and beverage industries (e.g., lipases, amylases)
Pharmaceutical and biotechnological applications (e.g., proteases, glycosidases)
Environmental science (e.g., esterases in pollutant degradation)
Clinical diagnostics and therapeutic enzyme development

Accurate measurement of hydrolase activity is critical to understanding enzyme specificity, efficiency, and potential industrial value.

Comprehensive Assay Services

How It Works

Step	Procedure	Details
1	Enzyme Preparation	Purification and preparation of hydrolase samples. Handling both crude extracts and recombinant enzymes.
2	Substrate Selection	Identification of appropriate natural or synthetic substrates. Consideration of substrate solubility, stability, and specificity.
3	Assay Development	Customized assays designed to match client requirements. Detection formats include: Spectrophotometric assays for absorbance-based monitoring. Fluorescent assays for sensitive, high-throughput detection. Chromatographic or titrimetric assays for detailed product analysis.
4	Enzyme Activity Determination	Real-time or endpoint measurement of bond hydrolysis. Rigorous quality control to ensure accuracy and reproducibility.
5	Data Analysis & Reporting	Comprehensive kinetic data, including parameters such as K_m, V_max, and k_cat. Expert interpretation to highlight substrate specificity and enzyme efficiency.

Contact Our Team

Explore Our Specialized Hydrolase Services

To address specific subclasses of hydrolases, Creative Enzymes offers focused services with optimized assays:

Enzyme Activity Measurement for Hydrolases Acting on Ester Bonds	This service specializes in enzymes such as esterases and lipases that cleave ester linkages. We provide robust assays to determine catalytic efficiency, substrate specificity, and reaction conditions—critical for industrial biocatalysis, food processing, and biofuel production.
Enzyme Activity Measurement for Glycosylases	Glycosylases catalyze the hydrolysis of glycosidic bonds in carbohydrates and glycoconjugates. Our assays offer precise characterization of their activity, providing valuable data for pharmaceutical research, metabolic engineering, and glycobiology.

Contact Our Team

Why Choose Creative Enzymes

Broad Expertise

Extensive experience across hydrolases acting on ester, glycosidic, peptide, and other bonds.

Advanced Technologies

Access to sensitive spectrophotometric, fluorescent, and chromatographic platforms.

Customizable Workflows

Flexible design to meet academic, industrial, or clinical research goals.

Reliable Data

Strict protocols and quality assurance ensure reproducibility and high accuracy.

Confidential Partnership

Client data and intellectual property are handled with the highest security.

Efficient Timelines

Streamlined workflows for timely delivery of results.

Representative Case Studies

Case 1: Magnetically Recoverable ZnFe₂O₄@MS for Lipase Immobilization

This study developed mesoporous zinc ferrite nanoparticles coated with amine-functionalized mesoporous silica (ZnFe₂O₄@MS) as an efficient nano-support for Candida rugosa lipase (CRL) immobilization. The hybrid system allowed easy magnetic recovery, high surface area, and strong covalent binding, nearly doubling enzyme activity (689 U/mg vs. 324 U/mg free). The immobilized enzyme-maintained stability at elevated temperatures, retaining >59% activity after five cycles. Additionally, ZnFe₂O₄@MS/CRL exhibited strong antibacterial effects, particularly against Staphylococcus aureus, and efficiently synthesized isoamyl acetate (banana flavor) with 64% yield. These results highlight its potential for sustainable, cost-effective biocatalysis and industrial applications.

Investigation of loading amount, biocatalytic activity, thermal and pH stabilities, and kinetic parameters of the enzymes of ZnFe2O4 Figure 1. Investigating chemical modifications and enzyme loading on the surface of ZnFe₂O₄@MS nanocomposite through FTIR spectra (A), the influence of the reaction medium pH (B) and temperature (C) on the stability and activity of free and immobilized lipase, and Lineweaver-Burk plot related to free and immobilized lipase enzyme (D). (Hosseinzadeh et al., 2024)

Case 2: Engineering Thermostable SS-BGL for Enhanced Ginsenoside Conversion

β-Glycosidase from Sulfolobus solfataricus (SS-BGL) is an efficient biocatalyst for producing compound K (CK) from protopanaxadiol ginsenosides. To improve its thermal stability, molecular dynamics simulations and dynamic cross-correlation mapping identified key binding residues and distal interacting networks. Rational mutagenesis targeting these sites yielded two optimized variants: SS-BGL-Q96E/N97D/N302D and SS-BGL-Q96E/N97D/N128D/N302D. These mutants exhibited 2.5- and 3.3-fold longer half-lives at 95 °C compared to the wild type, while maintaining or enhancing activity (161% and 116% relative ginsenoside conversion). This strategy demonstrates the power of computationally guided enzyme engineering for creating thermostable, high-performance biocatalysts.

Characterization of the enzymatic activity of wild-type and variants of S. solfataricus β-glycosidase Figure 2. Screening and enzymatic activity characterization of WT and variants. (A) Residual enzyme activity incubated at 95 °C for 30 min. (B) Relative enzymatic activity of variants with pNPG as substrate. (Shen et al., 2023)

FAQs

Q: Which types of hydrolases can you measure?

A: We provide activity measurement services for all subclasses of hydrolases, including esterases, lipases, proteases, glycosidases, nucleases, and more.
Q: What detection methods do you use?

A: Depending on enzyme type, we use spectrophotometric, fluorescent, chromatographic, or titrimetric assays, customized to client needs.
Q: Can you work with enzymes in crude extracts?

A: Yes, we can measure activity in crude extracts, partially purified, or purified samples depending on client requirements.
Q: Do you provide kinetic parameters in addition to activity measurements?

A: Yes, our services include determination of kinetic constants such as K_m, V_max, turnover number, and inhibition studies upon request.
Q: How long does a typical project take?

A: Most hydrolase activity measurement projects are completed within 2–4 weeks, depending on complexity.

References:

Hosseinzadeh H, Oveisi H, Meshkini A. Functionalized ZnFe2O4@Mesoporous silica nano-support for lipase enzyme immobilization: Enhanced biocatalysis and antibacterial activity for food industry applications. Food Bioscience. 2024;61:104985. doi:10.1016/j.fbio.2024.104985
Shen W, Dalby PA, Guo Z, Li W, Zhu C, Fan D. Residue effect-guided design: engineering of S. solfataricus β-glycosidase to enhance its thermostability and bioproduction of ginsenoside compound k. J Agric Food Chem. 2023;71(44):16669-16680. doi:10.1021/acs.jafc.3c04575

First Name:

Last Name:

Email *

Phone Number:

Company/Institution:

Country or Region:

Quantity:

Services & Products of Interested

Project Description:

For research and industrial use only, not for personal medicinal use.

Services

Online Inquiry