Advances in Enzyme Inhibition: Biophysical and Experimental Approaches

A special issue of Biophysica (ISSN 2673-4125).

Deadline for manuscript submissions: 31 August 2025 | Viewed by 388

Special Issue Editor

Special Issue Information

Dear Colleagues,

Enzymes are major targets in drug development due to their critical roles in biological processes. Investigating enzyme characteristics through purification, kinetic studies, stability profiling, and structural analysis is essential for understanding their functions and for guiding the design of more effective inhibitors. Both experimental and computational approaches—such as molecular docking and molecular dynamics—are crucial for achieving a comprehensive understanding.

This Special Issue, “Advances in Enzyme Inhibition: Biophysical and Experimental Approaches, aims to highlight recent progress in experimental and/or computational strategies for enzyme inhibition. Contributions that utilize biophysical methods or integrated approaches are especially encouraged.

Key words for the topics: Enzyme purification, enzyme stabilization, enzyme inhibitors, Mechanistic studies of enzyme activity, Structural and conformational analyses, computational modeling of enzyme-inhibitor interactions.

We invite original research and reviews that advance our understanding of enzyme inhibition.

Prof. Dr. Sung-Kun (Sean) Kim
Guest Editor

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Keywords

  • enzyme purification
  • enzyme stabilization
  • enzyme inhibitors
  • mechanistic studies of enzyme activity
  • structural and conformational analyses
  • computational modeling of enzyme-inhibitor interactions

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Published Papers (1 paper)

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Research

8 pages, 1000 KiB  
Article
A Novel Purification Process of Sardine Lipases Using Protein Ultrafiltration and Dye Ligand Affinity Chromatography
by Juan Antonio Noriega-Rodríguez, Armando Tejeda-Mansir and Hugo Sergio García
Biophysica 2025, 5(3), 35; https://doi.org/10.3390/biophysica5030035 - 10 Aug 2025
Viewed by 188
Abstract
Protein purification is often performed for various applications. However, enzyme purification processes typically involve multiple steps that reduce yield and increase production costs. To overcome these challenges, we developed a novel three-step process to purify a lipase from whole sardine viscera (WSV), leveraging [...] Read more.
Protein purification is often performed for various applications. However, enzyme purification processes typically involve multiple steps that reduce yield and increase production costs. To overcome these challenges, we developed a novel three-step process to purify a lipase from whole sardine viscera (WSV), leveraging protein properties and the structural affinity of lipases for dye ligands. A crude extract of the viscera (CEV) was obtained by grinding the whole viscera in 50 mM phosphate buffer (pH 7.0, Solution B) followed by centrifugation (6000× g; 30 min, 0 °C). Lipolytic activity (3.3 U/mg) was recorded only in the supernatant. The purification process began with ammonium sulfate fractionation (30–50% saturation), increasing lipolytic activity in the precipitate (PF30-50) to 32.9 U/mg. PF30-50 was then ultrafiltered using a 30 KDa MWCO membrane, where 5% of semi-purified lipases (SPLSV) was retained with an activity of 156.5 U/mg (UF30). Finally, the SPLSV was injected into a column packed with dye ligand affinity adsorbent, pre-equilibrated with 1.0 M ammonium sulfate in buffer A. The WSV lipase was eluted using a step gradient to progressively reduce salt concentration. SDS-PAGE analysis revealed a single band of purified lipase from sardine viscera (PLSV) corresponding to a molecular weight of 123.4 kDa, with a specific activity of 266.4 U/mg. The combination of ammonium sulfate precipitation, ultrafiltration, and dye-ligand affinity chromatography provides a scalable and reproducible approach with potential industrial relevance, particularly in biocatalysis and waste valorization contexts. Full article
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