Search Results (1,175)

Heterocyclic scaffolds are often present in many natural and non-natural products with important biological activity, such as synthetic intermediates used to synthesise many drugs. Among others, heterocycles based on a pyrimidine ring may have antioxidant, antibacterial, antiviral, antifungal, antituberculosis, and anti-inflammatory properties. The present study investigated commercially available microbial biocatalysts in the enzymatic desymmetrization reaction of bulky–bulky ketones derived from pyrimidine bases. The influence of some parameters on the efficiency of biocatalysis, i.e., the substrate concentration and pH of the reaction medium, was evaluated. In the one-step bioreduction catalysed by Saccharomyces cerevisiae, secondary alcohols with a defined absolute configuration were obtained with high enantiomeric excess up to 99% ee and moderate conversion. Biocatalysis offers economic and environmental benefits as an alternative to conventional methods, becoming a powerful tool in the synthesis of crowded alcohols. Full article

Extracellular electron transfer (EET) is key to the success of microbial fuel cells (MFCs). Clostridium sp. often occurs in MFC anode communities, but its ability to perform EET remains controversial. We have employed Clostridium pasteurianum DSM 525 as a biocatalyst in a glycerol-fed MFC, designated MFC_DSM. We have also followed the EET of this biocatalyst in the presence of a mediator, namely soluble neutral red (NR), soluble methyl viologen (MV), neutral red film (FNR), or methyl viologen film (FMV). MFC_DSM provided power and current densities (j) of 0.39 μW·cm⁻² and 2.47 μA·cm⁻², respectively, which evidenced that the biocatalyst performs direct electron transfer (DET). Introducing 150.0 µM NR or MV into the MFC_DSM improved the current density by 7.0- and 3.7-fold (17.05 and 8.45 μA·cm⁻²), respectively. After 20 cyclic voltammetry (CV) cycles, the presence of FNR in the MFC_DSM anodic chamber provided an almost twofold higher current density (30.76 µA·cm⁻²) compared to the presence of NR in the MFC_DSM. Introducing MV or FMV into the MFC_DSM anodic chamber gave practically the same current density after 10 CV cycles. The MFC_DSM anodic electrode might interact with FMV weakly than with FNR, so FNR is more promising to enhance C. pasteurianum DSM 525 EET within MFC_DSM. Full article

Since the 2005 discovery of the first enzyme capable of depolymerizing polyethylene terephthalate (PET), an aromatic polyester once thought to be enzymatically inert, extensive research has been undertaken to identify and engineer new biocatalysts for plastic degradation. This effort was directed toward developing efficient enzymatic recycling technologies that could overcome the limitations of mechanical and chemical methods. These enzymes are versatile molecules obtained from microorganisms living in various environments, including soil, compost, surface seawater, and extreme habitats such as hot springs, hydrothermal vents, deep-sea regions, and Antarctic seawater. Among various plastics, PET and polylactic acid (PLA) have been the primary focus of enzymatic depolymerization research, greatly enhancing our knowledge of enzymes that degrade these specific polymers. They often display unique catalytic properties that reflect their particular ecological niches. This review explores recent advancements in marine-derived enzymes that can depolymerize synthetic plastic polymers, emphasizing their structural and functional features that influence the efficiency of these catalysts in biorecycling processes. Current status and future perspectives of enzymatic plastic depolymerization are also discussed, with a focus on the underexplored marine enzymatic resources. Full article

The increasing disposal of emerging contaminants in the environment is a worldwide concern due to environmental impacts, such as toxicity, hormonal disorders, and bioaccumulation. The persistence of these pollutants in water bodies makes conventional pollutant removal techniques inefficient or partial, thus requiring the development of new, more effective, sustainable remediation technologies. Therefore, chitosan-based materials have emerged as a promising alternative for application in catalysis and contaminant removal. The biopolymer has functional properties that make it an excellent adsorbent capable of removing more specific pollutants, such as pharmaceuticals, microplastics, agricultural pesticides, and perfluoroalkyl and poly-fluoroalkyl substances, which are increasingly in evidence today. Therefore, this review of recent and advanced research into using chitosan to manufacture catalytic and adsorption materials offers an innovative approach to treating contaminants in aqueous environments, significantly reducing their presence and impact. It discusses the advantages of using chitosan as an adsorbent and catalyst and its role as a support for catalysts and biocatalysts. In addition, the review highlights the diversity of the physical forms of chitosan, such as particles, membranes, and hydrogels, and its possible chemical modifications, highlighting its effectiveness in catalytic applications and the removal of a wide range of emerging contaminants. Full article

Nucleoside phosphorylases (NPs) are pivotal enzymes in the salvage pathway, catalyzing the reversible phosphorolysis of nucleosides to produce nucleobases and α-D-ribose 1-phosphate. Due to their efficiency in catalyzing nucleoside synthesis from purine or pyrimidine bases, these enzymes hold significant industrial importance in the production of nucleoside-based drugs. Given that the thermodynamic equilibrium for purine NPs (PNPs) is favorable for nucleoside synthesis—unlike pyrimidine NPs (PyNPs, UP, and TP)—multi-enzymatic systems combining PNPs with PyNPs, UPs, or TPs are commonly employed in the synthesis of nucleoside analogs. In this study, we report the first development of two engineered bifunctional fusion enzymes, created through the genetic fusion of purine nucleoside phosphorylase I (PNP I) and thymidine phosphorylase (TP) from Thermus thermophilus. These fusion constructs, PNP I/TP-His and TP/PNP I-His, provide an innovative one-pot, single-step alternative to traditional multi-enzymatic synthesis approaches. Interestingly, both fusion enzymes retain phosphorolytic activity for both purine and pyrimidine nucleosides, demonstrating significant activity at elevated temperatures (60–90 °C) and within a pH range of 6–8. Additionally, both enzymes exhibit high thermal stability, maintaining approximately 80–100% of their activity when incubated at 60–80 °C over extended periods. Furthermore, the transglycosylation capabilities of the fusion enzymes were explored, demonstrating successful catalysis between purine (2′-deoxy)ribonucleosides and pyrimidine bases, and vice versa. To optimize reaction conditions, the effects of pH and temperature on transglycosylation activity were systematically examined. Finally, as a proof of concept, these fusion enzymes were successfully employed in the synthesis of various purine and pyrimidine ribonucleoside and 2′-deoxyribonucleoside analogs, underscoring their potential as versatile biocatalysts in nucleoside-based drug synthesis. Full article

The growing demand for sustainable chemical production has spurred significant interest in biocatalysis. This study is framed within the biocatalytic production of 1,3-dihydroxyacetone (DHA) from glycerol, a byproduct of biodiesel manufacturing. The main goal of this study is to address the challenge of identifying the optimal operating conditions. To achieve this, catalytic potential, a lumped parameter that considers both the activity and stability of immobilized biocatalysts, was used to guide the design of a multi-enzymatic system. The multi-enzymatic system comprises glycerol dehydrogenase (GlyDH) and NADH oxidase (NOX). The enzymatic oxidation of glycerol to DHA catalyzed by GlyDH requires the cofactor NAD+. The integration of NOX into a one-pot reactor allows for the in situ regeneration of NAD+, enhancing the overall efficiency of the process. Furthermore, immobilization on Ni⁺² agarose chelated supports, combined with post-immobilization modifications (glutaraldehyde crosslinking for GlyDH), significantly improved the stability and activity of both enzymes. The catalytic potential enabled the identification of the optimal operating conditions, which were 30 °C and pH 7.5, favoring NOX stability. This work establishes a framework for the rational design and optimization of multi-enzymatic systems. It highlights the crucial interplay between individual enzyme properties and process conditions to achieve efficient and sustainable biocatalytic transformations. Full article

Methylotrophic yeast Ogataea polymorpha BKM Y-2559 was immobilized in organosilicon sol–gel matrices using precursors isobutyltriethoxysilane (iBTES) and tetraethoxysilane (TEOS) to create an effective biocatalyst. The analytical and metrological performance of the biosensor permitted the determination of the optimum ratio of iBTES and TEOS, which was found to be 20/80 vol.%. The results of the scanning electron microscopy method demonstrated the formation of organosilicon material around microorganisms, as well as the ease with which metabolic products of yeast cells and substrates could diffuse through the obtained pores. A laboratory model of the biofilter was developed, exhibiting an oxidative capacity that varied from 0.14 to 1.25 gO₂/(m³ × cycle) in accordance with the initial level of water pollution and the degree of purification of moderately polluted water. The latter was found to be 20%, which aligns with the norm for drip biofilters operating in cyclic mode. Full article

This study explores the reasons behind the variations in the enantioselectivity of the sulfoxidation of methyl phenyl sulfide by marine-derived vanadium-dependent haloperoxidases (VHPOs). Twelve new VHPOs of marine organisms were overexpressed, purified, and tested for their ability to oxidize sulfide. Most of these marine enzymes exhibited nonenantioselective behavior, underscoring the uniqueness of AnVBPO from the brown seaweed Ascophyllum nodosum and CpVBPO from the red seaweed Corallina pilulifera, which produce (R)- and (S)-sulfoxides, respectively. The enantioselective sulfoxidation pathway is likely due to direct oxygen transfer within the VHPO active site. This was demonstrated through molecular docking and molecular dynamics simulations, which revealed differences in the positioning of sulfide within AnVBPO and CpVBPO, thus explaining their distinct enantioselectivities. Nonenantioselective VHPOs probably follow a different oxidation pathway, initiating with sulfide oxidation to form a positively charged radical. Further insights were gained from studying the catalytic effect of VO₄³⁻ on H₂O₂-driven sulfoxidation. This research improves the understanding of VHPO-mediated sulfoxidation and aids in developing biocatalysts for sulfoxide synthesis. Full article

The present study is integrated in a global effort to capitalize waste cooking oil (WCO) into versatile compounds by introducing an oxirane ring into the unsaturated carbon chain of fatty acid residues (the epoxidation of double bound). Therefore, an enzymatic method was set up for the epoxidation of artificially adulterated WCO (SFw) and WCO under real conditions (SFr) derived from sunflower biomass. Commercial lipase (Novozyme, NZ) was used as a biocatalyst for generating the peracid requested by the epoxidation pathway. Optimum experimental conditions (e.g., 1.5 wt% NZ, 1:1:0.5 = H₂O₂/double bonds/peracid precursor (molar ratio) and 12 h reaction time) allowed for the conversion of 90% of the SFw substrate into products with an oxirane ring. Octanoic acid was selected as the best peracid precursor. The versatility of the developed system was tested for olive, milk thistle, hemp and linseed oils as both fresh and WCO samples. The characterization of the oil samples before and after the enzymatic epoxidation allowed for the evaluation of the system performance. SFw/SFr exhibited a better susceptibility to enzymatic epoxidation. In addition, the reusability of the biocatalytic system was investigated. Furthermore, different strategies, such as biocatalyst coating and the addition of organic solvents/buffers were applied, limiting enzyme leaching, for the better recovery of the biocatalyst activity. Full article

Alginate lyase is an attractive biocatalyst that can specifically degrade alginate to produce oligosaccharides, showing great potential for industrial and medicinal applications. Herein, an alginate-degrading strain HB236076 was isolated from Sargassum sp. in Qionghai, Hainan, China. The low 16S rRNA gene sequence identity (<98.4%), ANI value (<71.9%), and dDDH value (<23.9%) clearly indicated that the isolate represented a potential novel species of the genus Vibrio. The genome contained two chromosomes with lengths of 3,007,948 bp and 874,895 bp, respectively, totaling 3,882,843 bp with a G+C content of 46.5%. Among 3482 genes, 3332 protein-coding genes, 116 tRNA, and 34 rRNA sequences were predicted. Analysis of the amino acid sequences showed that the strain encoded 73 carbohydrate-active enzymes (CAZymes), predicting seven PL7 (Alg1–7) and two PL17 family (Alg8, 9) alginate lyases. The extracellular alginate lyase from strain HB236076 showed the maximum activity at 50 °C and pH 7.0, with over 90% activity measured in the range of 30–60 °C and pH 6.0–10.0, exhibiting a wide range of temperature and pH activities. The enzyme also remained at more than 90% of the original activity at a wide pH range (3.0–9.0) and temperature below 50 °C for more than 2 h, demonstrating significant thermal and pH stabilities. Fe²⁺ had a good promoting effect on the alginate lyase activity at 10 mM, increasing by 3.5 times. Thin layer chromatography (TLC) and electrospray ionization mass spectrometry (ESI-MS) analyses suggested that alginate lyase in fermentation broth could catalyze sodium alginate to produce disaccharides and trisaccharides, which showed antimicrobial activity against Shigella dysenteriae, Aeromonas hydrophila, Staphylococcus aureus, Streptococcus agalactiae, and Escherichia coli. This research provided extended insights into the production mechanism of alginate lyase from Vibrio sp. HB236076, which was beneficial for further application in the preparation of pH-stable and thermo-stable alginate lyase and alginate oligosaccharides. Full article

Protein dynamics play a crucial role in biological function, encompassing motions ranging from atomic vibrations to large-scale conformational changes. Recent advancements in experimental techniques, computational methods, and artificial intelligence have revolutionized our understanding of protein dynamics. Nuclear magnetic resonance spectroscopy provides atomic-resolution insights, while molecular dynamics simulations offer detailed trajectories of protein motions. Computational methods applied to X-ray crystallography and cryo-electron microscopy (cryo-EM) have enabled the exploration of protein dynamics, capturing conformational ensembles that were previously unattainable. The integration of machine learning, exemplified by AlphaFold2, has accelerated structure prediction and dynamics analysis. These approaches have revealed the importance of protein dynamics in allosteric regulation, enzyme catalysis, and intrinsically disordered proteins. The shift towards ensemble representations of protein structures and the application of single-molecule techniques have further enhanced our ability to capture the dynamic nature of proteins. Understanding protein dynamics is essential for elucidating biological mechanisms, designing drugs, and developing novel biocatalysts, marking a significant paradigm shift in structural biology and drug discovery. Full article

Biodiesel has received tremendous attention as a sustainable energy source. This review presents an overview of various catalysts utilized in biodiesel production and compares their potential for producing biodiesel. Presented here are the excellent features of the various catalysts while highlighting their drawbacks. For instance, production of biodiesel with homogeneous base catalysts is easy but it can only be used with refined oils having low levels of free fatty acid (FFAs). When homogeneous acid is used in esterification, it causes reactor corrosion. Water and FFAs do not affect heterogeneous acid catalysts. Thus, transesterification of triglycerides into biodiesel and converting FFAs into biodiesel through esterification can be catalyzed more efficiently using a heterogeneous acid catalyst. Biocatalysts are also being used to produce biodiesel from oils with high FFAs. However, heterogeneous acid catalysts and biocatalysts are not suitable for industrial application due to serious mass transfer limitations. Biodiesel yield and conversion were compared over various catalysts in this paper. Also presented are the effects of different reaction parameters on biodiesel yield over different catalysts. The correct interplay of factors like reaction temperature, time, alcohol-to-oil molar ratio, and catalyst loading produces optimal process conditions that give the highest biodiesel yield. Full article

Phenylpropanoid sucrose esters are a large and important group of natural substances with significant therapeutic potential. This work describes a pilot study of the enzymatic hydroxycinnamoylation of sucrose and its derivatives which was carried out with the aim of obtaining precursors of natural phenylpropanoid sucrose esters, e.g., vanicoside B. In addition to sucrose, some chemically prepared sucrose acetonides and substituted 3′-O-cinnamates were subjected to enzymatic transesterification with vinyl esters of coumaric, ferulic and 3,4,5-trimethoxycinnamic acid. Commercial enzyme preparations of Lipozyme TL IM lipase and Pentopan 500 BG exhibiting feruloyl esterase activity were tested as biocatalysts in these reactions. The substrate specificity of the used biocatalysts for the donor and acceptor as well as the regioselectivity of the reactions were evaluated and discussed. Surprisingly, Lipozyme TL IM catalyzed the cinnamoylation of sucrose derivatives more to the 1′-OH and 4′-OH positions than to the 6′-OH when the 3′-OH was free and the 6-OH was blocked by isopropylidene. In this case, Pentopan reacted comparably to 1′-OH and 6′-OH positions. If sucrose 3′-O-coumarate was used as an acceptor, in the case of feruloylation with Lipozyme in CH₃CN, 6-O-ferulate was the main product (63%). Pentopan feruloylated sucrose 3′-O-coumarate comparably well at the 6-OH and 6′-OH positions (77%). When a proton-donor solvent was used, migration of the 3′-O-cinnamoyl group from fructose to the 2-OH position of glucose was observed. The enzyme hydroxycinnamoylations studied can shorten the targeted syntheses of various phenylpropanoid sucrose esters. Full article

This study explored the enantiocomplementary bioreduction of substituted 1-(arylsulfanyl)propan-2-ones in batch mode using four wild-type yeast strains and two different recombinant alcohol dehydrogenases from Lactobacillus kefir and Rhodococcus aetherivorans. The selected yeast strains and recombinant alcohol dehydrogenases as whole-cell biocatalysts resulted in the corresponding 1-(arylsulfanyl)propan-2-ols with moderate to excellent conversions (60–99%) and high selectivities (ee > 95%). The best bioreductions—in terms of conversion (>90%) and enantiomeric excess (>99% ee)—at preparative scale resulted in the expected chiral alcohols with similar conversion and selectivity to the screening reactions. Full article

The ever-increasing presence of micropollutants necessitates the development of environmentally friendly bioremediation strategies. Inspired by the remarkable versatility and potent catalytic activities of microbial enzymes, researchers are exploring their application as biocatalysts for innovative environmental cleanup solutions. Microbial enzymes offer remarkable substrate specificity, biodegradability, and the capacity to degrade a wide array of pollutants, positioning them as powerful tools for bioremediation. However, practical applications are often hindered by limitations in enzyme stability and reusability. Enzyme immobilization techniques have emerged as transformative strategies, enhancing enzyme stability and reusability by anchoring them onto inert or activated supports. These improvements lead to more efficient pollutant degradation and cost-effective bioremediation processes. This review delves into the diverse immobilization methods, showcasing their success in degrading various environmental pollutants, including pharmaceuticals, dyes, pesticides, microplastics, and industrial chemicals. By highlighting the transformative potential of microbial immobilized enzyme biocatalysts, this review underscores their significance in achieving a cleaner and more sustainable future through the mitigation of micropollutant contamination. Additionally, future research directions in areas such as enzyme engineering and machine learning hold immense promise for further broadening the capabilities and optimizing the applications of immobilized enzymes in environmental cleanup. Full article