Ancestral Sequence Reconstruction for Designing Biocatalysts and Investigating their Functional Mechanisms.

Biocatalysis has emerged as a green approach for efficient and sustainable production in various industries. In recent decades, numerous advancements in computational and predictive approaches, including ancestral sequence reconstruction (ASR) have sparked a new wave for protein engineers to improve and expand biocatalyst capabilities. ASR is an evolution-based strategy that uses phylogenetic relationships among homologous extant sequences to probabilistically infer the most likely ancestral sequences.

Multiple-Temperature-Responsive Double- and Triple-Network Hydrogels.

Temperature-responsive hydrogels which are reversibly activated attract much attention in the field of smart materials. Herein, double- and triple-network hydrogels exhibiting dual- or triple-volume transitions upon changes of temperature are fabricated. Interestingly, the incorporation of superhydrophilic polymer network leads to a significant difference swelling or shrinkage of the multiple-network hydrogels.

Mechanistic insights into allosteric regulation of the reductase component of -hydroxyphenylacetate 3-hydroxylase by -hydroxyphenylacetate: a model for effector-controlled activity of redox enzymes.

Understanding how an enzyme regulates its function through substrate or allosteric regulation is crucial for controlling metabolic pathways. Some flavin-dependent monooxygenases (FDMOs) have evolved an allosteric mechanism to produce reduced flavin while minimizing the use of NADH and the production of harmful hydrogen peroxide (HO). In this work, we investigated in-depth mechanisms of how the reductase component (C1) of -hydroxyphenylacetate (HPA) 3-hydroxylase (HPAH) from is allosterically controlled by the binding of HPA, which is a substrate of its monooxygenase counterpart (C2).

Spatiotemporal Retention of Structural Color and Induced Stiffening in Crosslinked Hydroxypropyl Cellulose Beads.

Hydroxypropyl cellulose (HPC) is known for its ability to form cholesteric liquid crystalline phases displaying vivid structural colors. However, these vibrant colors tend to fade over time when the material dries. This issue is a major bottleneck to finding practical applications for these materials.

CO fixation: cycloaddition of CO to epoxides using practical metal-free recyclable catalysts.

The conversion of CO into valuable chemicals is a crucial field of research. Cyclic organic carbonates have attracted great interest because they can be prepared under mild conditions and because of their structural versatility which enables a large variety of applications. Therefore, there is a need for potent and yet practical catalysts for the cycloaddition of CO to cyclic carbonates that are able to combine availability, low cost and an adequate performance.

Insights into the Synthesis of a Semiconducting Nickel Bis(dithiolene) Coordination Polymer.

Nickel bis(dithiolene) complexes are promising candidates for novel n-type semiconductors, which are air-stable and highly conductive. A key issue for further development is that their synthesis often yields undesired products, greatly limiting the degree of polymerization as well as purity and adversely affecting their electronic properties. Crucially, there is a lack of in-depth identification of these species and understanding of the reaction mechanism.

Economically Scalable Cu-based MOFs: Vital Role of Structural Integrity towards Selectivity on Azeotropic Ethanol Dehydration.

A facile, green, and economical method for the scalable synthesis of hydrophilic copper-triazole metal-organic frameworks (Cu-trz) is demonstrated. Numerous open metal sites within the highly crystalline porous structure of Cu-trz are generated through mild thermal activation, enabling its application in liquid-phase ethanol dehydration under ambient conditions. The frameworks with distinct crystallinity and particle sizes were achieved by modifying the synthesis process.

Abietane Diterpenoids from the Rhizomes of and Their Anti-Inflammatory Activities.

Six new abietane diterpenes, roscoeananes A-F (-), along with two known compounds (-) were isolated from the rhizomes of . The structures of all compounds were elucidated by analysis of spectroscopic data, and the absolute configurations were assigned by a comparison of the theoretical and experimental electronic circular dichroism (ECD) spectra and a comparison with literature values. The unreported compound is an ether-linked dimer of roscoeanane B ().

Unraveling the Nature of Interfacial Behavior in the LiTFSI-LiCl Aqueous Biphasic System.

Aqueous biphasic systems (ABSs) with water-in-salt electrolytes are gaining significant attention for their role in aqueous biphasic interphase studies, particularly in energy storage devices. Aqueous salt-salt biphasic electrolytes are considered a promising alternative to replace traditional liquid electrolytes commonly used in battery technologies, for example, membrane-less redox flow batteries, owing to their low cost and high ionic conductivity. However, the stability of the interphase over time must be considered, as it can impact the long-term electrochemical performance in various applications.

Integrating Genomic Data with the Development of CRISPR-Based Point-of-Care-Testing for Bacterial Infections.

Purpose Of Review: Bacterial infections and antibiotic resistance contribute to global mortality. Despite many infections being preventable and treatable, the lack of reliable and accessible diagnostic tools exacerbates these issues. CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)-based diagnostics has emerged as a promising solution.

Molecular models of PM6 for non-fullerene acceptor organic solar cells: How DAD and ADA structures impact charge separation and charge recombination.

The quadrupole moment of a non-fullerene acceptor (NFA) generated by the constituent electron donor (D) and acceptor (A) units is a significant factor that affects the charge separation (CS) and charge recombination (CR) processes in organic photovoltaics (OPVs). However, its impact on p-type polymer domains remains unclear. In this study, we synthesized p-type molecules, namely acceptor-donor-acceptor (ADA) and donor-acceptor-donor (DAD), which are components of the benchmark PM6 polymer (D: benzodithiophene and A: dioxobenzodithiophene).

Codon and Signal Peptide Optimization to Enhance Therapeutic Antibody Production from CHO Cells.

A simple and rapid method for reproducible transient expression experiments to quickly identify the best conditions before the development of a stable CHO cell line is described, using adalimumab as a model antibody. Firstly, the signal peptide (SP) is optimized using different in silico programs, of which those having a D score of more than 0.85 will be selected.

Pre-training strategy for antiviral drug screening with low-data graph neural network: A case study in HIV-1 K103N reverse transcriptase.

Graph neural networks (GNN) offer an alternative approach to boost the screening effectiveness in drug discovery. However, their efficacy is often hindered by limited datasets. To address this limitation, we introduced a robust GNN training framework, applied to various chemical databases to identify potent non-nucleoside reverse transcriptase inhibitors (NNRTIs) against the challenging K103N-mutated HIV-1 RT.

Nanodots functionalized with chitooligosaccharides for blocking chitoporins.

We report the synthesis of functionalized nanodots as potential powerful blockers of solute transport through a chitoporin. Ultrasmall silica nanocapsules with a diameter of ∼ 6 nm were coated with chitooligosaccharides to be used as a "lid" binding to the opening of the chitoporin VhChiP of Vibrio campbellii. Efficient blocking is attributed to the adequate size of the nanodots and their functionalization with oligochitosan, which has strong affinity towards the Vibrio chitoporin.

Designed functions of oxide/hydroxide nanosheets elemental replacement/doping.

Partial replacement of one structural element in a solid with another of a similar size was conducted to impart functionality to the solids and modify their properties. This phenomenon is found in nature in coloured gemstones and clay minerals and is used in materials chemistry and physics, endowing materials with useful properties that can be controlled by incorporated heteroelements and their amounts. Depending on the area of research (or expected functions), the replacement is referred to as "isomorphous substitution", "doping", Herein, elemental replacement in two-dimensional (2D) oxides and hydroxides (nanosheets or layered materials) is summarised with emphasis on the uniqueness of their preparation, characterisation and application compared with those of the corresponding bulk materials.

Enhancement of tryptophan 2-monooxygenase thermostability by semi-rational enzyme engineering: a strategic design to minimize experimental investigation.

Tryptophan 2-monooxygenase (TMO) is an FAD-bound flavoenzyme which catalyzes the oxidative decarboxylation of l-tryptophan to produce indole-3-acetamide (IAM) and carbon dioxide. The reaction of TMO is the first step of indole-3-acetic acid (IAA) biosynthesis. Although TMO is of interest for mechanistic studies and synthetic biology applications, the enzyme has low thermostability and soluble expression yield.

A 3D printed dual screen-printed electrode separation device for twin electrochemical mini-cell establishment.

We describe a tiny 3D-printed polymethyl-methacrylate-based plastic sleeve that houses two disposable screen-printed electrodes (SPE) and enables each of the working electrodes (WEs) to work independently, on a different side of a thin barrier, in its own electrochemical (EC) mini-cell, while the SPE counter and reference units are shared for electroanalysis. Optical and EC performance tests proved that the plastic divider between WE1 and WE2 efficiently inhibited solution mixing between the mini-cells. The two neighboring, independently operating mini-cells enabled matched differential measurements in the same sample solution, a tactic designed for elimination of electrochemical interference in complex samples.

Efficient dye removal using manganese oxide-modified nanocellulosic films from sugarcane bagasse.

Removing toxic dyes from industrial wastewater is crucial for environmental protection. This research introduced novel composite films of manganese oxide (MnO)-modified nanocellulose (MCel) and unmodified nanocellulose (Cel) derived from sugarcane bagasse for dye removal. Nanocellulose was extracted from sugarcane bagasse and subsequently transformed into MCel through in-situ MnO synthesis.

characterization techniques of protein corona around nanomaterials.

Nanoparticles (NPs) inevitably interact with proteins upon exposure to biological fluids, leading to the formation of an adsorption layer known as the "protein corona". This corona imparts NPs with a new biological identity, directly influencing their interactions with living systems and dictating their fates . Thus, gaining a comprehensive understanding of the dynamic interplay between NPs and proteins in biological fluids is crucial for predicting therapeutic effects and advancing the clinical translation of nanomedicines.

Glucose Oxidase-Based Glucose Biosensing with a Simple Dual Ag/AgCl Probe Conductivity Readout.

We describe a conductometric assay of the enzymatic conversion of glucose to gluconic acid by dissolved glucose oxidase (GOx), using the generation of proton and gluconate from the reaction product dissociation for glucose detection. Simple basics of ionic conductivity, a silver/silver chloride wire pair, and a small applied potential translate glucose-dependent GOx activity into a scalable cell current. Enzyme immobilization and complex sensor design, involving extra nanomaterials or microfabrication of electrode structures, are entirely avoided, in contrast to all modern electrochemical glucose biosensors.

An expanded molecular and systems toolbox for imaging, mapping, and controlling local translation.

Localized protein translation occurs through trafficking of mRNAs and protein translation machineries to different compartments of the cell, leading to rapid on-site synthesis of proteins in response to signaling cues. The spatiotemporally precise nature of the local translation process necessitates continual developments of technologies reviewed herein to visualize and map biomolecular components and the translation process with better spatial and temporal resolution and with fewer artifacts. We also discuss approaches to control local translation, which can serve as a design paradigm for subcellular genetic devices for eukaryotic synthetic biology.

Point-of-care CRISPR-based Diagnostics with Premixed and Freeze-dried Reagents.

Molecular diagnostics by Clustered Regularly Interspaced Short Palindromic Repeats (CRISPR)-based detection have high diagnostic accuracy and attributes that are suitable for use at point-of-care settings such as fast turnaround times for results, convenient simple readouts, and no requirement of complicated instruments. However, the reactions can be cumbersome to perform at the point of care due to their many components and manual handling steps. Herein, we provide a step-by-step, optimized protocol for the robust detection of disease pathogens and genetic markers with recombinase-based isothermal amplification and CRISPR-based reagents, which are premixed and then freeze-dried in easily stored and ready-to-use formats.