Thermostability Enhancement of Tagatose 4-Epimerase through Protein Engineering and Whole-Cell Immobilization

Journal of Agricultural and Food Chemistry, Journal Year: 2025, Volume and Issue: unknown

Published: Jan. 3, 2025

d-Tagatose, a rare sugar endowed with low-calorie property, superior taste quality, and probiotic functionality, has garnered significant research attention. However, the prevailing biological production methods relying on β-galactosidase l-arabinose isomerase face challenges including high cost suboptimal conversion efficiency. Consequently, it is of great significance to find efficient alternative routes for d-tagatose synthesis. Previously, Thermotoga petrophila tagaturonate 3-epimerase was modified function as tagatose 4-epimerase (T4E) enabling direct d-fructose d-tagatose. In this study, T4E further engineered through directed evolution, specifically targeting enhancement its thermostability application. This endeavor yielded promising variants superiority over those original enzyme. I430P exhibits half-life (t1/2) at 70 °C that 1.83-fold T4E, an increased melting temperature (Tm) 5.1 compared T4E. Additionally, G90S/T272A/I430P demonstrated 21.4% increase in specific activity At °C, t1/2 1.69-fold Tm 2.9 higher than Furthermore, whole-cell immobilization integrating these into robust biocatalytic system employed. innovative approach not only underscores practical feasibility modifying enzymes evolution but also establishes foundation cost-effective, large-scale

Language: Английский

---

Direct Electron Transfer–Type Oxidoreductases for Biomedical Applications

Annual Review of Biomedical Engineering, Journal Year: 2024, Volume and Issue: 26(1), P. 357 - 382

Published: March 1, 2024

Among the various types of enzyme-based biosensors, sensors utilizing enzymes capable direct electron transfer (DET) are recognized as most ideal. However, only a limited number redox DET with electrodes, that is, dehydrogenases harboring subunit or domain functions specifically to accept electrons from cofactor catalytic site and external acceptor. Such subunits domains act built-in mediators for between electrodes; consequently, such enable electrodes designated DET-type enzymes. fall into several categories, including cofactors reactions, by their protein hierarchic structures, oxidoreductases oligomeric structures subunits, monomeric domains. In this review, we cover science biomedical applications. First, introduce structural biology current understanding enzyme reactions. Next, describe recent technological developments based on applications, biosensors biochemical energy harvesting self-powered medical devices. Finally, after discussing how further engineer create enzymes, address future prospects in engineering.

Language: Английский

---

Rapid Prediction of Energy Level Alignment and Conductance of Single‐Molecule Junctions Through Intramolecular Dipole Moment

Advanced Functional Materials, Journal Year: 2024, Volume and Issue: unknown

Published: April 4, 2024

Abstract How substituents affect the conductance of single‐molecule junctions has been a subject ongoing debate. Here, comparative study on transport properties across broader range experimentally proposed and representative π‐conjugated molecular systems, each characterized by prototypical backbone motifs with distinct terminal groups featuring is conducted. Employing non‐equilibrium Green's function method within density functional theory, this investigation reveals significant impact junction due to asymmetric characteristics arising from both electronic spatial effects induced di‐substituents phenanthrene analogs C═C or B─N motifs. Remarkably, linear correlation uncovered, either negative positive, between dipole moment molecules conductance, contingent upon whether governed highest occupied orbital (HOMO) lowest unoccupied (LUMO) states. Furthermore, validity descriptor substantiated various substituted series, including 2,7‐dipyridylfluorene, oligo(phenylene ethynylene), 1,4‐diaminobenzene. These findings underscore pivotal role individual moment, easily determinable, in offering precise predictive insight into intricate junctions.

Language: Английский

---

Structural and electrochemical elucidation of biocatalytic mechanisms in direct electron transfer-type D-fructose dehydrogenase

Electrochimica Acta, Journal Year: 2024, Volume and Issue: 490, P. 144271 - 144271

Published: April 14, 2024

Language: Английский

---

Structural bioelectrochemistry of direct electron transfer-type multimeric dehydrogenases: Basic principle and rational strategies

Bioelectrochemistry, Journal Year: 2025, Volume and Issue: unknown, P. 108973 - 108973

Published: March 1, 2025

Language: Английский

---

Impacts of electrogenerated chemiluminescence mechanism on the emission spectra and intensities

Analytical Sciences, Journal Year: 2025, Volume and Issue: unknown

Published: Feb. 25, 2025

Abstract In this mini-review, recent progress in electrogenerated chemiluminescence (ECL) reported by our group is mainly introduced. First, the general principles of ECL and standard molecules for are described, then properties thermally activated delayed fluorescence (TADF) molecules, pyrrolopyrrole aza-BODIPYs (PPABs), pyrene (Py) its derivative, Eu(III) Tb(III) complexes The efficiency TADF increased spin up-conversion from T 1 to S state, that PPABs incremented kinetically due slower rate state formation than formation. excimer mechanism Py derivative discussed. It shown very sharp spectra can be obtained complexes. Furthermore, intense binary mixed systems using Ir(ppy) 3 electron donor or acceptor described. These results provide insight into how affects emission intensity, efficiency, spectrum. Graphical abstract

Language: Английский

---

Quantitative Elucidation of Catalytic Reaction of Truncated Aldehyde Dehydrogenase Based on Linear Free Energy Relationship

ACS Catalysis, Journal Year: 2025, Volume and Issue: unknown, P. 7283 - 7295

Published: April 18, 2025

Language: Английский

---

Enhancement of direct electron transfer by aromatic thiol modification with truncated d-fructose dehydrogenase

Electrochimica Acta, Journal Year: 2024, Volume and Issue: 502, P. 144804 - 144804

Published: Aug. 4, 2024

Language: Английский

---

Electrode Nanopatterning for Bioelectroanalysis and Bioelectrocatalysis

Electrochemistry, Journal Year: 2024, Volume and Issue: 92(2), P. 022005 - 022005

Published: Jan. 23, 2024

Although intensive research in the bioelectroanalysis domain last twenty years led to great improvements protein-based electrode and device performances, their large application remains limited by stability overtime under operational resting conditions.One under-studied issue is role played spatial distribution of enzymes on electrocatalysis.Actually, high fluxes metabolic pathways involve compartmentalization organization active biomolecules.In a mimicking way, it can be expected that controlled localization proteins surfaces may play overall electron transfer processes bioelectrocatalysis performances.In this short review, we will discuss recent developments surface patterning allowing tune manner density surface.We investigate how mixed functional layers, biological materials serve as protein platforms provide such patterning.

Language: Английский

---

The effect of number of layers of nanoporous gold films on their electrochemical behaviour

Electrochimica Acta, Journal Year: 2024, Volume and Issue: 489, P. 144233 - 144233

Published: April 7, 2024

Nanostructured material based electrodes are frequently used in electrochemical analysis and catalysis for their multifarious favourable properties. The deep interior surfaces of these are, however, not often well addressed due to diffusion constrains, leading a poor materials economy. present work demonstrates thickness control manipulation dealloyed nanoporous gold (NPG) using layer-by-layer method. viability the method is confirmed by electron microscopy characterisation. effect number NPG layers (from one five, thicknesses range 100-500 nm) on behaviour evaluated, 1) redox diffusing probe ferrocenemethanol, 2) dopamine undergoing proton-coupled two-electron transfer, 3) surface-confined osmium complex modified polymer, as 4) bioelectrocatalysis an enzyme, fructose dehydrogenase (FDH). Notably, results show that best performance achieved intermediate layers, suggesting tedious efforts fabricating deep/ thick nanostructures can be optimised.

Language: Английский

---