Enzyme immobilization can be carried out in either batch or flow mode. Developing flow-through enzymatic reactions requires integrating upstream flow bioprocesses, such as flow immobilization, to minimize downtime, reduce labor requirements, and support the development of sustainable bioprocesses. In this study, Burkholderia cepacia lipase (BCL) was immobilized on Immobead 150 P, and flow esterification of 5-hydroxymethylfurfural (HMF) with oleic acid was performed to synthesize 5-hydroxymethylfurfural oleate. For batch immobilization, the optimal enzyme loading was determined to be 7 mg protein per gram of support, yielding an activity yield (IY%) of 99.9% and a protein yield (IP%) of 98.6%. In flow mode, the best immobilization results were achieved at a flow rate of 0.05 mL/min, yielding an IY of 98% and an IP of 80% within 130 min. The resulting biocatalyst exhibited high operational stability, maintaining activity after five reaction cycles. Batch synthesis of 5-hydroxymethylfurfural oleate achieved conversions of 97% and 45% using 30 mM and 50 mM HMF, respectively, over 24 h. In flow esterification of HMF and oleic acid, conversions of 84% (30 mM HMF) and 75% (50 mM HMF) were obtained at a flow rate of 0.05 mL/min. The space-time yield (STY) was higher in flow bioreactors, with values of 4.7 mM/min and 5.3 mM/min at the same flow rate for 30 mM and 50 mM HMF, respectively. This work demonstrates the integration of two flow processeslipase immobilization and HMF esterification with oleic acidhighlighting the significant potential for process scale-up.

Up to date, the development of highly efficient, visible light-active catalysts remains a formidable challenge due to the enhanced rising of atmospheric CO2 concentration. This study discusses a class of ceria-based high-entropy oxides designed to optimize charge carrier dynamics, surface reactivity, and CO2 activation efficiency. Due to the advantages of high configurational entropy and multi-element synergy, these materials achieved improved photocatalytic performance, surpassing conventional ceria-based systems. Structural and spectroscopic analyses reveal that Pr3+/Pr4+ redox pairs and abundant oxygen vacancies create an electronically disordered yet thermodynamically stable environment, which enhances charge separation and suppresses electron-hole recombination. Photocatalytic experiments demonstrated that Ce0.2Zr0.2La0.2Pr0.2Sm0.2O2-δ (CZLPS) achieves the highest CO2 conversion rate, reaching a conversion of 20.3% under visible light irradiation, significantly surpassing pure ceria (1.4%), with a calculated space-time yield (STY) of 10.15 molCOkg-1h-1 under the same conditions. First-principles density functional theory (DFT) simulations were employed to investigate the CO2 reduction mechanism on CZLPS catalysts. The study elucidates the Gibbs free energy changes (ΔG) for each step of the reaction pathways leading to CO and HCOOH formation, highlighting the Zr site of CZLPS as the most active for the CO2RR, which is responsible for the outstanding catalytic activity.

In Cu-Zn catalysts used for the reverse water-gas shift (RWGS) reaction, Cu and Zn phases remain dispersed, while the Cu-Zn interphase is widely regarded as the catalytically active site. Herein, we engineered a ZnO-decorated Cu-rich Cu-Zn alloy using a mixed Cu/Zn oxalate-based supramolecular metal-organic framework precursor with compositionally homogeneous Cu/Zn mixing. The resulting Cu-Zn catalyst (Cu-Zn|C-oxo|R) maintained >99% CO selectivity and, at 550°C, delivered a Cu-normalized CO space-time yield (STY) of 1580 mmolCO gCu [-1] h[-1], higher than those of the Cu-only and commercial Cu-Zn-Al benchmarks. Under a high space velocity test, the Cu-Zn|C-oxo|R catalyst remained stable for >50 h at 550°C. Operando Cu K-edge x-ray absorption spectroscopy shows that Cu is reduced to a predominantly metallic local environment that persists under RWGS conditions. Complementary in situ x-ray diffraction and electron microscopy identify Cu-rich Cu-Zn alloy domains in contact with ZnO, while in situ diffuse reflectance infrared Fourier transform spectroscopy is consistent with an associative formate-mediated RWGS pathway at these interfaces. Density functional theory calculations further revealed that these interfacial sites favor the formate reaction pathway, providing a molecular-level rationale for the enhanced catalytic activity.

Copper-based catalysts suffer from sintering and excessive oxidation during the methanol oxidation carbonylation process to produce dimethyl carbonate (DMC), which leads to their easy deactivation and thus are restricted in industrial applications. In this work, we designed and fabricated a series of nitrogen-doped carbon-supported copper-cobalt nanoparticles (CuxCo-NC). A distinctive volcano-type dependence of catalytic activity on the Cu/Co molar ratio was revealed, and a remarkable DMC space-time yield (STY) of 5.35 g g-1 cat·h-1 was delivered by the optimal composition (Cu/Co = 3). There was a strong metal-support interaction between the Cu-Co nanoparticles and the N-doped carbon matrix, promoting the dispersion of the metal. This study has demonstrated the role of ZIF-derived nitrogen-doped carbon materials in the liquid-phase oxidation and carbonylation of methanol, providing a foundation for subsequent research.

Immobilized enzymes have become central to continuous biotransformations by enabling catalyst recovery, repeated utilization, simplified product separation, and stable long-term reactor operation. This study critically evaluates how immobilization strategies, support architectures, reactor configurations, and reaction media collectively govern enzyme stability, mass transfer, catalytic efficiency, and process productivity in continuous biocatalytic systems. Recent advances in adsorption, covalent immobilization, entrapment, encapsulation, cross-linked enzyme aggregates (CLEAs), membrane reactors, packed-bed reactors, monolithic reactors, and structured flow platforms are systematically assessed with emphasis on industrially relevant performance metrics. The analysis demonstrates that although immobilization substantially enhances operational stability and catalyst lifetime, overall process performance is dictated by the interplay between enzyme orientation, support microstructure, pore diffusion, substrate and product transport, water activity, cofactor availability, product inhibition, enzyme leaching, fouling, and hydrodynamic limitations. Despite significant progress, meaningful comparison across studies remains constrained by inconsistent reporting of key process parameters, including enzyme loading, immobilization efficiency, residence time, space-time yield (STY), turnover number (TON), operational half-life, catalyst productivity, pressure drop, and time-on-stream stability. The study identifies critical knowledge gaps in mechanistic understanding, standardized performance evaluation, and long-duration continuous operation. It proposes an integrated framework that combines rational immobilization design with transport modeling, in situ reaction diagnostics, and cofactor engineering.

Efficient enzymaticsynthesis of L-carnosine (L-Car) is often limited by competing hydrolysis and side reactions that reduce substrate utilization and product yield. In this study, a β-aminopeptidase (CbAP) from Caulobacter sp. was systematically engineered to enhance catalytic selectivity toward L-Car using β-alaninamide and L-histidine as substrates. Analysis of β-alaninamide-derived byproducts revealed two major competing pathways, namely β-alaninamide hydrolysis and self-ligation into short oligopeptides. To evaluate the balance between L-Car synthesis and competing reactions, a composite apparent selectivity index (RS) was introduced. Structure-guided mutagenesis targeting residues within 4-6 Å of the active site identified several improved variants. Iterative combination of beneficial mutations yielded the optimal mutant CbAPQ138FF96Y (CbAPM2), exhibiting an approximately 2.3-fold increase in RS relative to Q138F and an approximately 3.1-fold increase relative to the wild-type enzyme, with substantial suppression of undesired side reactions. Molecular dynamics simulations indicated that these improvements arise from reduced substrate binding stability and disrupted hydrogen-bonding interactions with β-alaninamide and L-Car. Under optimized 10 mL-scale conditions, CbAPM2 produced 37.3 mM L-Car within 1 h, corresponding to a 74.6% yield based on β-alanine methyl ester, with an STY of 8.44 g L[-] [1] h[-1].

This study aimed to produce extracellular lipases from Aspergillus niger using solid-state cultivation (SSC) of cottonseed bran and a blend of cottonseed bran and wheat bran (agroindustrial coproducts) as low-cost substrates. The highest activity (93.1 U/g dry mass) was achieved with cottonseed bran supplemented with castor oil at a moisture content of 54%. A mesoporous poly(styrene-co-divinylbenzene) (poly(Sty-co-DVB)) support (surface area of 259 m[2] g[-1]) was used for lipase immobilization. The immobilization yield ranged from 67.3% to 98.2%, whereas the biocatalyst's Ra was much lower, ranging from 5 to 30%. The heterogeneous biocatalysts were evaluated for the hydrolysis of castor oil, achieving ester-to-free fatty acid conversions ranging from 17.7% to 45%. Their performance was compared with that of commercial immobilized lipases (Lipozyme TL, Lipozyme RM, and Lipozyme 435). The SSC-derived biocatalysts showed activity comparable to the commercial ones. Furthermore, the heterogeneous biocatalyst maintained its catalytic efficiency over five consecutive hydrolysis reactions, with no significant decrease in conversion. Overall, this study demonstrates the feasibility of using agro-industrial by-products as inexpensive substrates for lipase production and highlights the potential of SSC-derived immobilized biocatalysts as sustainable, cost-effective alternatives for enzymatic hydrolysis.

Ultrasonic cavitation offers a powerful route to intensify leaching of refractory minerals, yet its effectiveness is often limited by persistent silica-rich transport restriction in silicate systems. Magnesium (Mg) extraction from serpentine is fundamentally constrained by incongruent dissolution and rapid formation of silica-rich alteration layers that suppress mass transport. Here, a coupled chemo‑sonic activation strategy is developed to elucidate how fluoride-mediated silicate destabilization and ultrasound‑driven transport intensification jointly redirect residue evolution toward transport-accessible pore architecture. Comparative leaching experiments were conducted under ambient, thermal, fluoride‑assisted, ultrasonic, and combined operating modes. Conventional acid leaching reached only ∼20% Mg[2+] extraction, while thermal leaching accelerated early kinetics but plateaued near ∼50% due to silica‑controlled diffusion resistance. Fluoride activation destabilized silica-rich alteration domains, increasing extraction to >80%, whereas ultrasound-assisted leaching alone achieved near‑thermal performance at <40 °C, consistent with enhanced interfacial renewal. Synergistically, their coupled operation induced a non‑linear enhancement, reaching ∼86% extraction within 60 min at low temperature using reduced fluoride dosage. Space-time yield (STY) analysis confirmed genuine process intensification (20.23 g(Mg).L[-1].h[-1]) rather than time‑shifted conversion. Integrated structural and textural characterization demonstrates that enhanced leachability arises from suppression of silica densification and the formation of a transport‑accessible pore architecture rather than surface‑area maximization. These findings demonstrate that chemo‑sonic activation suppresses silica-rich transport restriction and redirects residue evolution toward a transport-accessible, wide-mesopore-dominated architecture, enabling high Mg[2+] extraction under low-temperature conditions.

CO2 is a major greenhouse gas as well as an abundant, cheap, and renewable carbon resource. Synthesis of ethanol from CO2 using a homogeneous catalyst represents a promising protocol for CO2 utilization, yet achieving high catalytic performance with a cost-effective catalyst remains a challenge. Herein, we report a highly efficient Ru-Co-PPh3 bimetallic catalyst for ethanol synthesis using CO2, methanol, and H2. The ethanol space time yield (STY) reached as high as 4.4 g L[-1] h[-1], with a turnover frequency (TOF) of 23.2 h[-1] and an ethanol selectivity of 85.6 C-mol% in total products, outperforming the previous reports of homogeneous catalytic systems for conversion of CO2 to ethanol. The PPh3 ligand enabled the high-efficiency reaction by direct utilization of much cheaper Ru-Co precursors and with lower catalyst dosage. Notably, the PPh3 ligand played multiple roles in improving the activity, selectivity and stability of the catalyst. The reaction mechanism was also discussed based on a series of control experiments. This work provides a sustainable and practical way of ethanol synthesis using CO2 as a feedstock.

Satellite DNA is a dynamic component of plant genomes and a valuable source of cytogenetic markers, but its diversity and chromosomal distribution in polyploid Triticeae remain insufficiently studied. Here, low-coverage whole-genome sequencing, graph-based repeat clustering, quantitative PCR, multivariate statistics and fluorescence in situ hybridization (FISH) were used to identify and characterize satellite repeats in Elymus and related Triticeae species. Sixteen repeat clusters (E1-E16), with monomer lengths of 118-667 bp, showed distinct taxonomic distributions and copy-number profiles across 14 species. Correlation analysis, principal component analysis and hierarchical clustering revealed concerted variation among repeats and separated the perennial taxa Elymus and Pseudoroegneria from Triticum, Secale, Hordeum and Dasypyrum. Spearman correlation analysis identified E7 and E9 as putative candidates associated with St/StY genomic backgrounds, whereas E10 was identified as a putative candidate associated with the H genome. These statistical associations require independent cytogenetic validation. Contrasting copy numbers of E6 and E11 in bread wheat cv. Chinese Spring versus Dasypyrum villosum (L.) Candargy identified them as V-genome candidates. FISH localized E6 to the terminal regions of chromosomes 3VL, 4VS and 7VS, and E11 to 4VL. Karyotyping further revealed that two lines previously considered as wheat-D. villosum addition lines were in fact substitution lines: W3 was identified as a 3V(3D) substitution line and W4 as a 4V(4B) substitution line, whereas W7 retained its 7V addition status. These results expand the set of chromosomal markers for comparative genomics and introgression analysis in wheat.

The bottom-up strategy shows great potential to build up environmentally benign gasoline-range isoparaffins directly from syngas. Herein, we report that Fe-substituted MFI zeolite could significantly enhance the selective formation of gasoline in comparison to the conventional MFI and extra-framework Fe-modified MFI. CO conversion reaches 42.5% and C5-C11 selectivity 77.0%, corresponding to a space-time-yield (STY) 10.8 mmolC/(gcat·h) with CH4 selectivity only 1.1% at a space velocity of 3000 mL/(g·h). Characterizations with UV-vis spectroscopy (UV-vis), UV-Raman spectroscopy, x-ray absorption spectroscopy (XAS), magic-angle-spinning (MAS) nuclear magnetic resonance (NMR), H2 temperature-programmed reduction (H2-TPR), and Fourier-transform infrared spectroscopy (FTIR) demonstrate that ferric species are atomically dispersed within the MFI framework, generating Si─OH─Fe Brønsted acid sites. Butene model reactions and theoretical calculations reveal that the enhanced activity could be attributed to the facilitated methylation of olefins with ketene. This study provides fundamental understanding of the chain growth process in OXZEO catalysis and offers guidance for the rational design of catalysts for the production of high-quality gasoline from syngas.

Salmonella Typhi (STy), the causative agent of typhoid fever, preferentially infects the ileum, but the basis for this preference has remained unknown. We show that STy exploits MUC2-derived monosaccharides, such as N-acetylgalactosamine (GalNAc), to upregulate invasion and target the ileum-opposite to the response of closely related S. Typhimurium (STm), whose invasion is inhibited by MUC2. Three STy-specific extracellular residues of the outer-membrane protein OmpW (K141/T142/Q193) engage GalNAc, and a periplasmic relay through the short FepE_STy, RS12510, and TonB transmits this signal to activate the master invasion regulator HilD. Mice lacking MUC2 or gut microbiota lose STy ileal tropism, which is restored by oral GalNAc-containing monosaccharides. An OmpW triple-mutant STy strain establishes a murine oral STy infection model, identifying STy-MUC2 glycan interactions as an early step of typhoid pathogenesis and a potential target for prevention.

The class Bunyaviricetes encompasses several highly pathogenic viruses that cause lethal hemorrhagic fevers. Due to their limited prevention and treatment options and high pathogenicity, these viruses require handling in biosafety level-4 facilities. Within the bunyaviruses, arenaviruses are particularly notable for their pathogenicity and ability to cause severe hemorrhagic disease in humans. The cap-dependent endonuclease (CEN) is a unique and crucial enzyme involved in the replication cycle of these viruses. As humans do not possess a similar enzyme, CEN represents an ideal target for antiviral drug development with reduced risk of side effects. Recently, we identified a promising CEN inhibitor (CENi) demonstrating potent inhibition of virus replication. In this manuscript, we demonstrate the successful therapeutic efficacy of CENis against Lassa fever and Argentine hemorrhagic fever virus infections in guinea pig models of lethal hemorrhagic fever. In addition, we identified several CENis with antiviral activity against other highly pathogenic arenaviruses. These findings further support the potential of CENis as therapeutic agents for arenavirus infections that cause severe and often lethal hemorrhagic fever. Collectively, our results suggest that CENis are promising candidates for pan-arenavirus therapy and may also have broader utility against other CEN-containing viruses for which no approved antiviral treatments currently exist.IMPORTANCEArenaviruses, such as Lassa virus and Junin virus, cause severe hemorrhagic fevers in humans and are associated with high mortality rates and limited treatment options. Because of their pathogenicity and potential for outbreaks, these viruses represent an important global health threat. The viral cap-dependent endonuclease (CEN) plays a critical role in arenavirus replication by enabling the cap-snatching process required for viral mRNA synthesis. Notably, mammalian cells lack a comparable enzyme, making CEN an attractive target for antiviral drug development. In this study, we demonstrate that inhibitors targeting the arenavirus CEN effectively suppress viral replication and provide therapeutic protection in animal models of lethal Lassa virus and Junin virus infections. We also identify compounds with antiviral activity against multiple highly pathogenic arenaviruses. These findings establish CEN inhibition as a promising strategy for the development of broad-spectrum antivirals against arenaviruses that cause life-threatening hemorrhagic fevers.