Enzyme Activity Measurement for Transferases

Transferases are a vital class of enzymes that catalyze the transfer of functional groups between molecules, driving a wide range of biological processes from metabolism to biosynthesis. Accurate activity measurement of transferases is essential for understanding their roles, optimizing their industrial applications, and advancing enzyme engineering. Creative Enzymes offers reliable, precise, and customized Enzyme Activity Measurement Services for Transferases, providing clients with high-quality data and expert insights to support both research and applied projects.

Why Is Enzyme Activity Measurement for Transferases Matter

Transferases are responsible for transferring functional groups such as amino, phosphate, methyl, glycosyl, and acyl groups from one substrate to another. These enzymes are essential in biochemical pathways like energy metabolism, signal transduction, protein modification, and drug metabolism. Because their function is often highly specific, measuring the activity of transferases requires careful selection of substrates, optimized assay conditions, and rigorous data analysis.

Applications of transferase activity measurement include:

• Drug discovery and pharmacological studies
• Enzyme engineering and directed evolution
• Biocatalysis for chemical and pharmaceutical industries
• Metabolic pathway engineering and synthetic biology

Comprehensive Service Offerings

How It Works

Explore Our Specialized Transferase Activity Measurement Services

To provide more targeted support for your research and industrial applications, Creative Enzymes offers specialized services within the transferase family. Each service is designed with optimized assay systems and tailored workflows to capture the unique activity of different subclasses of transferases.

Enzyme Activity Measurement for Phosphorus Transferases

Phosphorus transferases, including kinases and phosphotransferases, are central to energy metabolism and signal transduction. Our service uses sensitive assays—such as spectrophotometric and fluorescent methods—to accurately quantify phosphate group transfer. These measurements provide insights into enzyme kinetics, substrate specificity, and regulatory roles, supporting applications in drug discovery, metabolic engineering, and synthetic biology.

Enzyme Activity Measurement for Glycosyl, Hexosyl, and Pentosyl Transferases

This service focuses on enzymes that catalyze sugar group transfers, essential for glycosylation, polysaccharide biosynthesis, and carbohydrate metabolism. We develop customized assays to measure transferase activity with precision, revealing key information on enzyme efficiency, sugar donor specificity, and reaction rates. The results are valuable for pharmaceutical research, industrial biocatalysis, and glycobiology studies.

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Why Chose Creative Enzymes

Case Studies and Real-World Applications

Case 1

Case 1: Catalytic activity profile of polyP:AMP phosphotransferase from Myxococcus xanthus

Myxococcus xanthus produces polyphosphates (polyPs) during starvation, with polyphosphate kinase (Ppk) driving fruiting body formation. This study characterized polyP:AMP phosphotransferase (Pap), a class II Ppk2 enzyme. Pap activity was enhanced by Mn²⁺/Mg²⁺, showing higher affinity for long-chain polyPs but maximum catalytic efficiency with polyP60–70. Interestingly, Pap sequentially generated ADP and ATP from AMP, revealing additional polyP:ADP phosphotransferase activity. Starvation increased Pap activity 2.3–2.4-fold, and recombinant Pap cooperated with Ppk1 or adenylate kinase (AdkA) to produce ATP from AMP, ADP, and polyP. These findings highlight Pap's crucial role in energy regeneration during nutrient limitation.

Figure 1. Effects of reaction conditions on Pap activity. (A–C) Effects of pH (A), temperature (B), and metal ions (C) on ADP synthesis by Pap. These reactions were performed as described in Materials and methods. The data are expressed as the percentage of Pap activity relative to that at pH 8.0 (A), at 50 °C (B), or in the presence of 5 mM Mn²⁺ (C), set as 100%. (D) Substrate specificity of Pap. (Kimura and Kamatani, 2021)

Case 2

Case 2: YM155 Inhibits NleB and SseK Arginine Glycosyltransferase Activity

Type III secretion system effectors NleB and SseK are glycosyltransferases that modify protein substrates on arginine residues. A high-throughput screen of 42,498 compounds identified YM155 (sepantronium bromide) as an inhibitor of E. coli NleB1, Citrobacter rodentium NleB, and Salmonella enterica SseK1/2. YM155 showed no toxicity to mammalian cells or cross-reactivity with host O-linked N-acetylglucosaminyltransferase. In cell assays, YM155 reduced Salmonella survival in mouse macrophage-like cells without affecting bacterial growth, highlighting its potential as a selective anti-virulence agent and mechanistic probe to study NleB/SseK-mediated glycosylation in enteric pathogens.

Figure 2. Primary screening results. (A) Distribution of Z' scores across all plates in NleB1 inhibition assays. An average Z'-score of 0.77 +/− 0.08 was obtained. (B) High-throughput screening (HTS) results. (Zhu et al., 2021)

FAQs

Q: What types of transferases can you measure?

A: We work with all subclasses of transferases, including glycosyltransferases, kinases, methyltransferases, acyltransferases, aminotransferases, and more.

Q: Which assay formats do you provide for activity measurement?

A: Depending on enzyme type and project requirements, we use spectrophotometric, fluorescent, electrochemical, or radioisotope-based assays.

Q: Can you measure activity in crude extracts, or is purification required?

A: Both options are available. While purification is recommended for precise measurements, we can design assays for crude or partially purified enzyme preparations if needed.

Q: How long does a typical project take?

A: Most transferase activity measurement projects are completed within 2–4 weeks, depending on assay complexity and enzyme type.

Q: Do you provide kinetic analysis (K_m, V_max, etc.) in addition to activity measurement?

A: Yes, we provide complete kinetic profiling, including K_m, V_max, turnover numbers, and inhibition studies if requested.

Q: Do you provide enzyme activity measurement for other enzyme types?

A: Yes. In addition to transferases, we offer comprehensive activity measurement services for all major enzyme classes, including oxidoreductases, hydrolases, lyases, isomerases, and ligases.