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Hot Biological Catalysis: Isothermal Titration Calorimetry to Characterize Enzymatic Reactions

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PEI-Treated Eggshell and Onion Membranes as Natural Supports for Urease Immobilization: A Performance and Stability

Abdullah Said Okuşluk¹, Abdulhalim Kılıç¹

¹Department of Molecular Biology and Genetics, Istanbul Technical University, 34469 Istanbul, Turkey.

|November 10, 2025

Summary

This summary is machine-generated.

Immobilizing Jack Bean urease (JBU) on waste-derived eggshell (ESM) and onion membranes (OM) enhances stability and reduces costs. ESM offers better enzyme loading and reusability, while OM provides superior operational stability.

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Area of Science:

Biotechnology
Biomaterials Science
Enzyme Engineering

Background:

Jack Bean urease (JBU) possesses significant biotechnological potential but faces challenges with stability and recovery costs in its free form.
Immobilization onto sustainable, waste-derived materials like eggshell membrane (ESM) and onion membrane (OM) presents a viable solution.

Purpose of the Study:

To systematically evaluate and compare the immobilization of JBU onto PEI-pretreated ESM and OM.
To assess the morphology, enzyme kinetics, stability, and heavy metal resistance of the immobilized JBU systems.

Main Methods:

Scanning Electron Microscopy (SEM) for morphological analysis.
Enzyme kinetic studies to determine apparent K_m and V_max.
Stability assessments including thermal, dry storage, reusability, and resistance to Cu²⁺ inhibition.

Main Results:

ESM exhibited higher enzyme loading due to its fibrous structure compared to OM.
Immobilization led to increased apparent K_m and V_max, with OM showing a greater increase in V_max.
OM-JBU demonstrated enhanced thermal (up to 80 °C) and dry storage stability.
ESM-JBU offered superior reusability, retaining 50% activity after 4 cycles.
Both immobilized systems showed significant protection against Cu²⁺ inhibition, retaining over 70% activity.

Conclusions:

A performance trade-off exists: ESM excels in enzyme loading and reusability, while OM offers better operational stability.
Waste-derived membranes (ESM and OM) are effective, cost-efficient platforms for developing robust immobilized enzyme systems.
These findings highlight the potential of utilizing waste biomaterials for advanced biotechnological applications.