Related Experiment Video
Updated: Jun 3, 2025

Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions
Published on: April 4, 2014
Emergence of ADM-mediated bioconjugate to enhance longevity and catalytic efficiency of urease
Shailja Rai1, Poorn Prakash Pande1, Krishna Kumar2
1Department of Chemistry and Environmental Science, Madan Mohan Malaviya University of Technology, Gorakhpur 273010, Uttar Pradesh, India.
Abstract:
The versatile nature of the urease enzyme makes it a valuable asset in biological and industrial contexts. The creation of bioconjugates using enzyme-polymer combinations has extended the shelf life and stability of urease. A triblock copolymer, PAM-co-PDPA-co-PMAA@urease (ADM@urease), was synthesized using acrylamide (AM), 2,5-dioxopyrrolidin-1-ylacrylate (DPA), methacrylic acid (MAA), and urease via the RAFT-Grafting-To polymerization method. This polymeric interface stabilizes the enzyme and enhances substrate binding and product release, significantly boosting enzymatic efficiency. To enhance pH's influence on urease activity, three ADM grades were developed by adjusting pH-responsive MAA levels, confirmed by GPC analysis. ADM micellized at acidic pH values of 6.47 or lower, with a critical micelle concentration (CMC) of at least 0.125 mg/mL. Kinetic evaluations using Berthelot reagents at various pH levels and temperatures compared free enzyme and urease encapsulated in ADM@urease. The Michaelis-Menten constant (Km) values, derived from the Lineweaver-Burk plot, were similar for both forms. The ADM@urease demonstrated optimal stability and catalytic efficacy with a Km value of 1.18 and Vmax of 1.92 at pH 4. By improving the stability, efficiency, and performance of urease, this encapsulation technology offers potential for sustainable, eco-friendly industrial applications and advancements in biotechnology.
Related Concept Videos
Introduction to Mechanisms of Enzyme Catalysis
Enzymes
Enzyme deficiencies can often translate into life-threatening diseases. For example, a genetic abnormality resulting in the deficiency of the enzyme G6PD...
Catalytically Perfect Enzymes
Most enzymes...
Olefin Metathesis Polymerization: Acyclic Diene Metathesis (ADMET)
Similar to cross-metathesis, ADMET also involves the formation of metallacyclobutane intermediate by [2+2] cycloaddition of one of the double bonds of a terminal diene with...
Introduction to Enzymes
Most enzymes are proteins that speed up biochemical reactions without being consumed. Enzymes contain one or more active sites that...
Allosteric Proteins-ATCase
Aspartate transcarbamoylase (ATCase) is a cytosolic enzyme that catalyzes the condensation of L-aspartate and carbamoyl phosphate to N-carbamoyl-L-aspartate. This reaction is the first step in pyrimidine biosynthesis. UTP and CTP, the end products of the pyrimidine synthesis...

