Evidence of CT complex formation between a probe and unreacted methylimidazole in imidazolium cation based ionic liquids: sensing by functionalised 2-benzyledinemalononitrile.

Here we report for the first time that unreacted N-methylimidazole (MIM) which is often a common impurity, can corrupt the results of a photophysical investigation of a probe molecule in imidazolium cation based RTILs. In addition, we propose a new type of sensor (functionalised 2-benzyledinemalononitrile derivative) for easy colorimetric detection and quantitative estimation of unreacted colourless MIM in RTILs.

pH-rate profiles of L-arabinitol 4-dehydrogenase from Hypocrea jecorina and its application in L-xylulose production.

l-Arabinitol 4-dehydrogenase (LAD) from Hypocrea jecorina (HjLAD) was cloned and overexpressed in Escherichia coli BL21 (DE3). The kinetics of l-arabinitol oxidation by NAD(+), catalyzed by HjLAD, was studied within the pH range of 7.0-9.5 at 25°C. The turnover number (kcat) and the catalytic efficiency (kcat/Km) were 4200min(-1) and 290mM(-1)min(-1), respectively. HjLAD showed the highest turnover number and catalytic efficiency among all previously characterized LADs. In further application of HjLAD, rare l-sugar l-xylulose was produced by the enzymatic oxidation of arabinitol to give a yield of approximately 86%.

Immobilization of L-arabinitol dehydrogenase on aldehyde-functionalized silicon oxide nanoparticles for L-xylulose production.

L-Xylulose is a potential starting material for therapeutics. However, its translation into clinical practice has been hampered by its inherently low bioavailability. In addition, the high cost associated with the production of L-xylulose is a major factor hindering its rapid deployment beyond the laboratory. In the current study, L-arabinitol 4-dehydrogenase from Hypocrea jecorina (HjLAD), which catalyzes the conversion of L-arabinitol into L-xylulose, was immobilized onto various carriers, and the immobilized enzymes were characterized. HjLAD covalently immobilized onto silicon oxide nanoparticles showed the highest immobilization efficiency (94.7 %). This report presents a comparative characterization of free and immobilized HjLAD, including its thermostability and kinetic parameters. The thermostability of HjLAD immobilized on silicon oxide nanoparticles was more than 14.2-fold higher than free HjLAD; the t1/2 of HjLAD at 25 °C was enhanced from 190 min (free) to 45 h (immobilized). In addition, the immobilized HjLAD retained 94 % of its initial activity after 10 cycles. When the immobilized HjLAD was used to catalyze the biotransformation of L-arabinitol to L-xylulose, 66 % conversion and a productivity of 7.9 g · h(-1) · L(-1) were achieved. The enhanced thermostability and reusability of HjLAD suggest that immobilization of HjLAD onto silicon oxide nanoparticles has the potential for use in the industrial production of rare sugars.

Role of surface residue 184 in the catalytic activity of NADH oxidase from Streptococcus pyogenes.

Nicotinamide adenine dinucleotide (NADH) oxidase from Streptococcus pyogenes (SpNox) is a flavoprotein harboring one molecule of noncovalently bound flavin adenine dinucleotide. It catalyzes the oxidation of NADH by reducing molecular O2 to H2O directly through a four-electron reduction. In this study, we selected the lysine residues on the surface of SpNox and mutated them into arginine residues to study the effect on the enzyme activity. A single-point mutation (K184R) at the surface of SpNox enhanced NADH oxidase activity by approximately 50 % and improved thermostability with 46.6 % longer half life at 30 °C. Further insights into the function of residue K184 were obtained by substituting it with other nonpolar, polar, positively charged, and negatively charged residues. To elucidate the role of this residue, computer-assisted molecular modeling and substrate docking were performed. The results demonstrate that even a single mutation at the surface of the enzyme induces changes in the interaction at the active site and affects the activity and stability. Additionally, the data also suggest that the K184R mutant can be used as an effective biocatalyst for NAD(+) regeneration in L-rare sugar production.

CARDPSoML: Comparative approach to analyze and predict cardiovascular disease based on medical report data and feature fusion approach.

Background and Aims: Diabetes patients are at high risk for cardiovascular disease (CVD), which makes early identification and prompt management essential. To diagnose CVD in diabetic patients, this work attempts to provide a feature-fusion strategy employing supervised learning classifiers. Methods: Preprocessing patient data is part of the method, and it includes important characteristics connected to diabetes including insulin resistance and blood glucose levels. Principal component analysis and wavelet transformations are two examples of feature extraction techniques that are used to extract pertinent characteristics. The supervised learning classifiers, such as neural networks, decision trees, and support vector machines, are then trained and assessed using these characteristics. Results: Based on the area under the receiver operating characteristic curve, sensitivity, specificity, and accuracy, these classifiers' performance is closely evaluated. The assessment findings show that the classifiers have a good accuracy and area under the receiver operating characteristic curve value, suggesting that the suggested strategy may be useful in diagnosing CVD in patients with diabetes. Conclusion: The recommended method shows potential as a useful tool for developing clinical decision support systems and for the early detection of CVD in diabetes patients. To further improve diagnostic skills, future research projects may examine the use of bigger and more varied datasets as well as different machine learning approaches. Using an organized strategy is a crucial first step in tackling the serious problem of CVD in people with diabetes.

Role of conserved glycine in zinc-dependent medium chain dehydrogenase/reductase superfamily.

The medium-chain dehydrogenase/reductase (MDR) superfamily consists of a large group of enzymes with a broad range of activities. Members of this superfamily are currently the subject of intensive investigation, but many aspects, including the zinc dependence of MDR superfamily proteins, have not yet have been adequately investigated. Using a density functional theory-based screening strategy, we have identified a strictly conserved glycine residue (Gly) in the zinc-dependent MDR superfamily. To elucidate the role of this conserved Gly in MDR, we carried out a comprehensive structural, functional, and computational analysis of four MDR enzymes through a series of studies including site-directed mutagenesis, isothermal titration calorimetry, electron paramagnetic resonance (EPR), quantum mechanics, and molecular mechanics analysis. Gly substitution by other amino acids posed a significant threat to the metal binding affinity and activity of MDR superfamily enzymes. Mutagenesis at the conserved Gly resulted in alterations in the coordination of the catalytic zinc ion, with concomitant changes in metal-ligand bond length, bond angle, and the affinity (K(d)) toward the zinc ion. The Gly mutants also showed different spectroscopic properties in EPR compared with those of the wild type, indicating that the binding geometries of the zinc to the zinc binding ligands were changed by the mutation. The present results demonstrate that the conserved Gly in the GHE motif plays a role in maintaining the metal binding affinity and the electronic state of the catalytic zinc ion during catalysis of the MDR superfamily enzymes.

Molecular cloning and characterization of a GH11 endoxylanase from Chaetomium globosum, and its use in enzymatic pretreatment of biomass.

An endo-1,4-ß-xylanase gene, xylcg, was cloned from Chaetomium globosum and successfully expressed in Escherichia coli. The complete gene of 675 bp was amplified, cloned into the pET 28(a) vector, and expressed. The optimal conditions for the highest activity of the purified recombinant XylCg were observed at a temperature of 40 °C and pH of 5.5. Using oat-spelt xylan, the determined K m, V max, and k cat/K m values were 0.243 mg ml⁻¹, 4,530 U mg⁻¹ protein, and 7,640 ml s⁻¹ mg⁻¹, respectively. A homology model and sequence analysis of XylCg, along with the biochemical properties, confirmed that XylCg belongs to the GH11 family. Rice straw pretreated with XylCg showed 30 % higher conversion yield than the rice straw pretreated with a commercial xylanase. Although xylanases have been characterized from fungal and bacterial sources, C. globosum XylCg is distinguished from other xylanases by its high catalytic efficiency and its effectiveness in the pretreatment of lignocellulosic biomass.

From protein engineering to immobilization: promising strategies for the upgrade of industrial enzymes.

Enzymes found in nature have been exploited in industry due to their inherent catalytic properties in complex chemical processes under mild experimental and environmental conditions. The desired industrial goal is often difficult to achieve using the native form of the enzyme. Recent developments in protein engineering have revolutionized the development of commercially available enzymes into better industrial catalysts. Protein engineering aims at modifying the sequence of a protein, and hence its structure, to create enzymes with improved functional properties such as stability, specific activity, inhibition by reaction products, and selectivity towards non-natural substrates. Soluble enzymes are often immobilized onto solid insoluble supports to be reused in continuous processes and to facilitate the economical recovery of the enzyme after the reaction without any significant loss to its biochemical properties. Immobilization confers considerable stability towards temperature variations and organic solvents. Multipoint and multisubunit covalent attachments of enzymes on appropriately functionalized supports via linkers provide rigidity to the immobilized enzyme structure, ultimately resulting in improved enzyme stability. Protein engineering and immobilization techniques are sequential and compatible approaches for the improvement of enzyme properties. The present review highlights and summarizes various studies that have aimed to improve the biochemical properties of industrially significant enzymes.

Advancements in lanthanide-based perovskite oxide semiconductors for gas sensing applications: a focus on doping effects and development.

Lanthanide-based perovskite oxide semiconductors have garnered significant attention due to their exceptional electrical and sensing properties, making them promising candidates for gas sensing applications. This review paper focuses on developments and the impact of doping in lanthanide-based perovskite oxide semiconductors for gas sensing purposes. The review explores the factors influencing gas sensing performance, such as operating temperature, dopant selection, and target gas species. The role of dopants in enhancing gas sensing selectivity, sensitivity, response/recovery times, and stability is discussed in detail. Comparisons are drawn between doped perovskite oxide semiconductors, undoped counterparts, and other gas-sensing materials. Practical applications of lanthanide-based perovskite oxide semiconductor gas sensors are outlined, including environmental monitoring, industrial process control, and healthcare. The review also identifies current challenges and future perspectives in the field, such as the exploration of novel doping strategies and integration with emerging technologies like the Internet of Things (IoT). The findings emphasize the potential of these materials in advancing gas sensing technology and the importance of continued research in this field.

Molecular determinants of the cofactor specificity of ribitol dehydrogenase, a short-chain dehydrogenase/reductase.

Ribitol dehydrogenase from Zymomonas mobilis (ZmRDH) catalyzes the conversion of ribitol to d-ribulose and concomitantly reduces NAD(P)(+) to NAD(P)H. A systematic approach involving an initial sequence alignment-based residue screening, followed by a homology model-based screening and site-directed mutagenesis of the screened residues, was used to study the molecular determinants of the cofactor specificity of ZmRDH. A homologous conserved amino acid, Ser156, in the substrate-binding pocket of the wild-type ZmRDH was identified as an important residue affecting the cofactor specificity of ZmRDH. Further insights into the function of the Ser156 residue were obtained by substituting it with other hydrophobic nonpolar or polar amino acids. Substituting Ser156 with the negatively charged amino acids (Asp and Glu) altered the cofactor specificity of ZmRDH toward NAD(+) (S156D, [k(cat)/K(m)(,NAD)]/[k(cat)/K(m)(,NADP)] = 10.9, where K(m)(,NAD) is the K(m) for NAD(+) and K(m)(,NADP) is the K(m) for NADP(+)). In contrast, the mutants containing positively charged amino acids (His, Lys, or Arg) at position 156 showed a higher efficiency with NADP(+) as the cofactor (S156H, [k(cat)/K(m)(,NAD)]/[k(cat)/K(m)(,NADP)] = 0.11). These data, in addition to those of molecular dynamics and isothermal titration calorimetry studies, suggest that the cofactor specificity of ZmRDH can be modulated by manipulating the amino acid residue at position 156.

Probing the role of sigma π interaction and energetics in the catalytic efficiency of endo-1,4-ß-xylanase.

Chaetomium globosum endo-1,4-ß-xylanase (XylCg) is distinguished from other xylanases by its high turnover rate (1,860 s(-1)), the highest ever reported for fungal xylanases. One conserved amino acid, W48, in the substrate binding pocket of wild-type XylCg was identified as an important residue affecting XylCg's catalytic efficiency.

Characterization of H2O-forming NADH oxidase from Streptococcus pyogenes and its application in l-rare sugar production.

A nicotinamide adenine dinucleotide (NADH) oxidase from Streptococcus pyogenes MGAS10394 (SpNox) was cloned and overexpressed in Escherichia coli BL21 (DE3). The purified SpNox enzyme had optimal pH and temperature of 7.0 and 55°C, respectively, with a K(m) of 27.0µM and a k(cat)/K(m) of 1.1×10(7)s(-1)M(-1). SpNox showed the highest activity among all known NADH oxidases, and site-directed mutagenesis and docking analysis shed light on the molecular basis of its unusually high activity. The characteristics of SpNox may prove to be useful for NAD(+) regeneration in the production of l-rare sugar.

Mechanistic studies on the flavin:NADH reductase (PrnF) from Pseudomonas fluorescens involved in arylamine oxygenation.

We report the mechanistic studies of a FAD:NADH reductase (PrnF) involved in arylamine oxygenation. PrnF catalyzes the reduction of FAD via a sequential ordered bi-bi mechanism with NADH as the first substrate to bind and FADH(2) as the first product to be released. The residues Asp145 and His146 are proposed as catalytic acid/base residues for PrnF based on pH profile and molecular dynamics simulation studies. These studies provide the first detailed account of the mechanism of the flavin reductase involved in arylamine oxygenation.

Characterization of a recombinant aryl ß-glucosidase from Neosartorya fischeri NRRL181.

An isolated gene from Neosartorya fischeri NRRL181 encoding a ß-glucosidase (BGL) was cloned, and its nucleotide sequence was determined. DNA sequence analysis revealed an open reading frame of 1,467 bp, capable of encoding a polypeptide of 488 amino acid residues. The gene was over-expressed in Escherichia coli, and the protein was purified using nickel-nitrilotriacetic acid chromatography. The purified recombinant BGL showed a high level of catalytic activity, with V (max) of 886 µmol min(-1) mg-protein(-1) and a K (m) of 68 mM for p-nitrophenyl-ß-D: -glucopyranoside (pNPG). The optimal temperature for enzyme activity was about 40°C, and the optimal pH was about 6.0. A homology model of N. fischeri BGL1 was constructed based on the X-ray crystal structure of Phanerochaete chrysosporium BGLA. Molecular dynamics simulation studies of the enzyme with the pNPG and cellobiose shed light on the unique substrate specificity of N. fischeri BGL1 only towards pNPG.

Transition metal oxides as a cathode for indispensable Na-ion batteries.

The essential requirement to harness well-known renewable energy sources like wind energy, solar energy, etc. as a component of an overall plan to guarantee global power sustainability will require highly efficient, high power and energy density batteries to collect the derived electrical power and balance out variations in both supply and demand. Owing to the continuous exhaustion of fossil fuels, and ever increasing ecological problems associated with global warming, there is a critical requirement for searching for an alternative energy storage technology for a better and sustainable future. Electrochemical energy storage technology could be a solution for a sustainable source of clean energy. Sodium-ion battery (SIB) technology having a complementary energy storage mechanism to the lithium-ion battery (LIB) has been attracting significant attention from the scientific community due to its abundant resources, low cost, and high energy densities. Layered transition metal oxide (TMO) based materials for SIBs could be a potential candidate for SIBs among all other cathode materials. In this paper, we discussed the latest improvement in the various structures of the layered oxide materials for SIBs. Moreover, their synthesis, overall electrochemical performance, and several challenges associated with SIBs are comprehensively discussed with a stance on future possibilities. Many articles discussed the improvement of cathode materials for SIBs, and most of them have pondered the use of Na x MO2 (a class of TMOs) as a possible positive electrode material for SIBs. The different phases of layered TMOs (Na x MO2; TM = Co, Mn, Ti, Ni, Fe, Cr, Al, V, and a combination of multiple elements) show good cycling capacity, structural stability, and Na+ ion conductivity, which make them promising cathode material for SIBs. This review discusses and summarizes the electrochemical redox reaction, structural transformations, significant challenges, and future prospects to improve for Na x MO2. Moreover, this review highlights the recent advancement of several layered TMO cathode materials for SIBs. It is expected that this review will encourage further development of layered TMOs for SIBs.

Correction: Transition metal oxides as a cathode for indispensable Na-ion batteries.

[This corrects the article DOI: 10.1039/D2RA03601K.].

Enhancement of CO gas sensing performance by Mn-doped porous ZnSnO3 microspheres.

This work reports the synthesis of Mn-doped ZnSnO3 microspheres (Zn1-x Mn x SnO3) using a simple co-precipitation method with (x = 0 to 0.15) and characterized for structural, morphological, surface area, and sensing properties. X-ray diffraction (XRD) analysis revealed the face-centered cubic structure of Mn-doped ZnSnO3 samples. Brunauer-Emmett-Teller (BET) analysis demonstrated the variation in surface area from 15.229 m2 g-1 to 42.999 m2 g-1 with x = 0 to 0.15 in Zn1-x Mn x SnO3. XPS indicates the change in the defect levels by Mn doping, which plays a crucial role in chemical sensors. Indeed a significant increase (≈311.37%) in CO gas sensing response was observed in the x = 0.10 sample compared to pure ZnSnO3 with a simultaneous reduction in operating temperature from 250 to 200 °C. Moreover, remarkable enhancements in response/recovery times (≈6.6/34.1 s) were obtained in the x = 0.10 sample. The Mn-doped ZnSnO3 could be a promising candidate for CO gas sensing devices used for maintaining air quality.

Further biochemical studies on aminopyrrolnitrin oxygenase (PrnD).

Active site modeling of dimerization interface in combination with site-directed mutagenesis indicates that the electron in the PrnD Rieske oxygenase can be transferred by either of two pathways, one involving Asp183' and the other involving Asn180'. In addition, the overexpression of the isc operon involved in the assembly of iron-sulfur clusters increased the catalytic activity of PrnD in Escherichia coli by a factor of at least 4.

Covalent immobilization of recombinant Rhizobium etli CFN42 xylitol dehydrogenase onto modified silica nanoparticles.

Rare sugars have many applications in food industry, as well as pharmaceutical and nutrition industries. Xylitol dehydrogenase (XDH) can be used to synthesize various rare sugars enzymatically. However, the immobilization of XDH has not been performed to improve the industrial production of rare sugars. In this study, silica nanoparticles which have high immobilization efficiency were selected from among several carriers for immobilization of recombinant Rhizobium etli CFN42 xylitol dehydrogenase (ReXDH) and subjected to characterization. Among four different chemical modification methods to give different functional groups, the silica nanoparticle derivatized with epoxy groups showed the highest immobilization efficiency (92%). The thermostability of ReXDH was improved more than tenfold by immobilization on epoxy-silica nanoparticles; the t(1/2) of the ReXDH was enhanced from 120 min to 1,410 min at 40 °C and from 30 min to 450 min at 50 °C. The K(m) of ReXDH was slightly altered from 17.9 to only 19.2 mM by immobilization. The immobilized ReXDH had significant reusability, as it retained 81% activity after eight cycles of batch conversion of xylitol into L-xylulose. A∼71% conversion and a productivity of 10.7 g h(-1)l(-1) were achieved when the immobilized ReXDH was employed to catalyze the biotransformation of xylitol to L-xylulose, a sugar that has been used in medicine and in the diagnosis of hepatitis. These results suggest that immobilization of ReXDH onto epoxy-silica nanoparticles has potential industrial application in rare sugar production.