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The Characterization and Application of Enzymes in Bioprocesses

A special issue of International Journal of Molecular Sciences (ISSN 1422-0067). This special issue belongs to the section "Biochemistry".

Deadline for manuscript submissions: 20 March 2025 | Viewed by 1789

Special Issue Editor


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Guest Editor
Research Institute on Terrestrial Ecosystems-IRET-CNR, Via Pietro Castellino 111, 80131 Naples, Italy
Interests: protein biochemistry; enzyme and protein purification; enzyme characterization; extremozymes; extremophilic microorganisms; biotechnology for sustainability; biocatalysts; biorefinery; biomass and waste valorization; bioresources; bioactive molecules for well-being

Special Issue Information

Dear Colleagues,

Enzymes are protein catalysts that are found in all forms of organisms and play a crucial role in several bioprocesses; this is a vital area of biotechnology that utilizes living organisms and their enzymes to produce various types of products. The aim of biocatalysts is to accelerate the speed and efficiency of chemical reactions with high substrate specificity and without being consumed. In recent years, enzymes and biological molecules have garnered increased research interest due to the importance and versatility of their utilization. They are employed as catalysts in a variety of fields, and for their application in the technological, medical, environmental and industrial domains, they require an enhanced capacity in order to fulfil the operating conditions of such processes. To determine the feasibility of applying enzymes in various fields, they must be characterized, and their function and application studied via multidisciplinary approaches. The objective of this Special Issue is to collect original research articles, reviews, communications, and so on addressing the study, characterization and application of biocatalysts of different origins. Studies on the bioprocessing, biotechnological design, optimization and application (diagnostic/medical, industrial, agro-industrial, environmental, etc.) of biocatalysts are welcome.

Dr. Loredana Marcolongo
Guest Editor

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Keywords

  • biocatalysts
  • enzymatic characterization
  • enzymatic action
  • protein engineering
  • biocatalysts application
  • bioprocessing

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Published Papers (1 paper)

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Research

20 pages, 3533 KiB  
Article
Exploring the Kinetics and Thermodynamics of a Novel Histidine Ammonia-Lyase from Geobacillus kaustophilus
by Francisco Manuel Salas-Garrucho, Alba Carrillo-Moreno, Lellys M. Contreras, Felipe Rodríguez-Vico, Josefa María Clemente-Jiménez and Francisco Javier Las Heras-Vázquez
Int. J. Mol. Sci. 2024, 25(18), 10163; https://doi.org/10.3390/ijms251810163 - 21 Sep 2024
Viewed by 1231
Abstract
Histidine ammonia-lyase (HAL) plays a pivotal role in the non-oxidative deamination of L-histidine to produce trans-urocanic, a crucial process in amino acid metabolism. This study examines the cloning, purification, and biochemical characterization of a novel HAL from Geobacillus kaustophilus (GkHAL) [...] Read more.
Histidine ammonia-lyase (HAL) plays a pivotal role in the non-oxidative deamination of L-histidine to produce trans-urocanic, a crucial process in amino acid metabolism. This study examines the cloning, purification, and biochemical characterization of a novel HAL from Geobacillus kaustophilus (GkHAL) and eight active site mutants to assess their effects on substrate binding, catalysis, thermostability, and secondary structure. The GkHAL enzyme was successfully overexpressed and purified to homogeneity. Its primary sequence displayed 40.7% to 43.7% similarity with other known HALs and shared the same oligomeric structure in solution. Kinetic assays showed that GkHAL has optimal activity at 85 °C and pH 8.5, with high thermal stability even after preincubation at high temperatures. Mutations at Y52, H82, N194, and E411 resulted in a complete loss of catalytic activity, underscoring their essential role in enzyme function, while mutations at residues Q274, R280, and F325 did not abolish activity but did reduce catalytic efficiency. Notably, mutants R280K and F325Y displayed novel activity with L-histidinamide, expanding the substrate specificity of HAL enzymes. Circular dichroism (CD) analysis showed minor secondary structure changes in the mutants but no significant effect on global GkHAL folding. These findings suggest that GkHAL could be a promising candidate for potential biotechnological applications. Full article
(This article belongs to the Special Issue The Characterization and Application of Enzymes in Bioprocesses)
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