Engineering Cellular Metabolism.

Metabolic engineering is the science of rewiring the metabolism of cells to enhance production of native metabolites or to endow cells with the ability to produce new products. The potential applications of such efforts are wide ranging, including the generation of fuels, chemicals, foods, feeds, and pharmaceuticals. However, making cells into efficient factories is challenging because cells have evolved robust metabolic networks with hard-wired, tightly regulated lines of communication between molecular pathways that resist efforts to divert resources. Here, we will review the current status and challenges of metabolic engineering and will discuss how new technologies can enable metabolic engineering to be scaled up to the industrial level, either by cutting off the lines of control for endogenous metabolism or by infiltrating the system with disruptive, heterologous pathways that overcome cellular regulation.

MXene-mediated reconfiguration induces robust nickel-iron catalysts for industrial-grade water oxidation.

Nickel-iron oxy/hydroxides (NiFeOxHy) emerge as an attractive type of electrocatalysts for alkaline water oxidation reaction (WOR), but which encounter a huge challenge in stability, especially at industrial-grade large current density due to uncontrollable Fe leakage. Here, we tailor the Fe coordination by a MXene-mediated reconfiguration strategy for the resultant NiFeOxHy catalyst to alleviate Fe leakage and thus reinforce the WOR stability. The introduction of ultrafine MXene with surface dangling bonds in the electrochemical reconfiguration over Ni-Fe Prussian blue analogue induces the covalent hybridization of NiFeOxHy/MXene, which not only accelerates WOR kinetics but also improves Fe oxidation resistance against segregation. As a result, the NiFeOxHy coupled with MXene exhibits an extraordinary durability at ampere-level current density over 1,000 h for alkaline WOR with an ultralow overpotential of only 307 mV. This work provides a broad avenue and mechanistic insights for the development of nickel-iron catalysts toward industrial applications.

Two-dimensional constrained chaos and industrial revolution cycles.

Since the 1760s, at least three industrial revolutions have occurred. To explain this phenomenon, we introduce two-dimensional (2D) constrained chaos. Using a model of innovation dynamics, we show that an industrial-revolution-like technology burst, driven by investment/saving motives for R&D activities, recurs about every one hundred years if the monopolistic use of a new technology lasts about 8 y.

Lewis acid-dominated aqueous electrolyte acting as co-catalyst and overcoming N2 activation issues on catalyst surface.

The growing demands for ammonia in agriculture and transportation fuel stimulate researchers to develop sustainable electrochemical methods to synthesize ammonia ambiently, to get past the energy-intensive Haber-Bosch process. However, the conventionally used aqueous electrolytes limit N2 solubility, leading to insufficient reactant molecules in the vicinity of the catalyst during electrochemical nitrogen reduction reaction (NRR). This hampers the yield and production rate of ammonia, irrespective of how efficient the catalyst is. Herein, we introduce an aqueous electrolyte (NaBF4), which not only acts as an N2-carrier in the medium but also works as a full-fledged "co-catalyst" along with our active material MnN4 to deliver a high yield of NH3 (328.59 µg h-1 mgcat-1) at 0.0 V versus reversible hydrogen electrode. BF3-induced charge polarization shifts the metal d-band center of the MnN4 unit close to the Fermi level, inviting N2 adsorption facilely. The Lewis acidity of the free BF3 molecules further propagates their importance in polarizing the N≡N bond of the adsorbed N2 and its first protonation. This push-pull kind of electronic interaction has been confirmed from the change in d-band center values of the MnN4 site as well as charge density distribution over our active model units, which turned out to be effective enough to lower the energy barrier of the potential determining steps of NRR. Consequently, a high production rate of NH3 (2.45 × 10-9 mol s-1 cm-2) was achieved, approaching the industrial scale where the source of NH3 was thoroughly studied and confirmed to be chiefly from the electrochemical reduction of the purged N2 gas.

Tandem Dual-Site PbCu Electrocatalyst for High-Rate and Selective Glycine Synthesis at Industrial Current Densities.

Direct electrosynthesis of high-value amino acids from carbon and nitrogen monomers remains a challenge. Here, we design a tandem dual-site PbCu electrocatalyst for efficient amino acid electrosynthesis. Using oxalic acid (H2C2O4) and hydroxylamine (NH2OH) as the raw reactants, for the first time, we have realized the flow-electrosynthesis of glycine at the industrial current density of 200 mA cm-2 with Faradaic efficiency over 78%. In situ ATR-FTIR spectroscopy characterizations reveal a favorable tandem pathway on the dual-site catalyst. Specifically, the Pb site drives the highly selective electroreduction of H2C2O4 to form glyoxylic acid, and the Cu site accelerates the fast hydrogenation of oxime to form a glycine product. A glycine electrosynthesis (GES)-formaldehyde electrooxidation (FOR) assembly is further established, which synthesizes more valuable chemicals (HCOOH, H2) while minimizing energy consumption. Altogether, we introduce a new strategy to enable the one-step electrosynthesis of high-value amino acid from widely accessible monomers.

Assessment of growth, and ion uptake of plant species, Conocarpus erectus and Dodonaea viscosa, on industrial solid waste.

Present study assessed the growth of two plant species and ion uptake by them grown on different proportion of industrial solid waste and garden soil. The industrial waste having high concentration of chemicals were used with garden soil at different proportion i.e. 0% (T0), 5% (T1), 10% (T2), 15% (T3) and 20% (T4). Two species namely Conocarpus erectus (alien plant) and Dodonaea viscosa (indigenous) were used as test plants in pot study. Different parameters including growth, physiology, and anatomy of plants and concentration of cations (Na+, K+, Ca2+, and Mg2+) in the plant shoot and root were measured at different time duration (initial, 1st, 2nd, 3rd and 4th month). The key objective of the study was to use these plants to establish their plantations on the barren lands where industrial solid wastes were being disposed of. C. erectus showed better growth than D. viscosa, as well as more uptake of ions. A significant increase in plant growth was observed in fourth month in T1, where plant height reached 24.5% and 46% for C. erectus and D. viscosa, respectively. At harvest, in C. erectus, no significant difference in the fresh (65-78 g) and dry weight (24-30 g) of the shoot was observed across treatments compared to the control. In D. viscosa, at the time of harvest, the fresh and dry weights of the root and shoot showed a strong, significantly decreasing pattern across T1, T2, and T3, leading to the death of the plant at T3 and T4. Further, optimum ratio of waste soil to garden soil was found as 10:90 and 20:80 to establish the plantations of D. viscosa and C. erectus, respectively in areas where such solid waste from industries are disposed. Findings can be used for the restoration of such solid waste for the sustainable management of industrial areas and their associated ecosystems.

Quality assessment of flax advanced breeding lines varying in seed coat color and their potential use in the food and industrial applications.

BACKGROUND: With the increasing consumer awareness of the strong relationship between food and health, flax became a promising functional food due to its bioactive nutraceutical composition. Intra-specific crosses of eight contrasting flax genotypes were performed previously, and within segregating F6 progeny families, we investigated a close-up composition of phytochemicals derived from whole seeds. RESULTS: The considerable genetic variation among the flax F6 families suggested that intra-specific hybridization is essential in flax breeding to obtain and broaden genetic variability and largely affirmed the opportunity for selecting promising lines. Also, significant variations in the targeted metabolite contents and antioxidant properties were observed among brown and yellow-seeded families. Notably, brown-seeded families expressed the highest average values of saturated fatty acids, protein, fiber, tocopherol, phenolics, SDG, and SECO lignans. Yellow-seeded families represented the highest average content of unsaturated fatty acids and mucilage. The cultivation year significantly affects flaxseed's composition and functional properties, presumably due to temperature, humidity, and sunshine time differences. Interestingly, the seeds obtained in warmer conditions were more potent and had more chemical constituents. The favorable genetic correlations among all evaluated traits suggest the possibility of joint genetic selection for several nutritional and phytochemical characteristics in flax. The current study highlights the importance and utilization of 19 top families as their seeds and oil play imperative roles in the pharmaceuticals and food industries. The antioxidant capacity of the seeds showed that families 84B, 23B, 35Y, 95Y, 30B, 88B, and 78B serve as a natural source of dietary antioxidants beneficial to human health. To increase the oxidative stability of the flaxseed oil, the quality evaluation identified some families with low levels of linolenic acid. CONCLUSIONS: These findings are essential to improving flaxseed's nutritional quality and therapeutic properties through a bulk breeding program.

Dual-Functional Ru/Ni-B-P Electrocatalyst Toward Accelerated Water Electrolysis and High-Stability.

Development of advanced electrocatalysts for the green hydrogen production by water electrolysis is an important task to reduce the climate and environmental issues as well as to meet the future energy demands. Herein, Ru/Ni-B-P sphere electrocatalyst is demonstrated by a combination of hydrothermal and soaking approaches, meeting the industrial requirement of low cell voltage with stable high-current operation. The Ru/Ni-B-P sphere catalyst demonstrates low overpotentials of 191 and 350 mV at 300 mA cm-2 with stable high current operation, ranking it as one of the best oxygen evolution reaction (OER) electrocatalysts. The bifunctional 2-E system demonstrates a low cell voltage of 2.49 V at 2000 mA cm-2 in 6 m KOH at 60 °C of harsh industrial operation condition. It also demonstrates outstanding stability with continuous 120 h (5 days) CA operation at 1000 mA cm-2. Further, the hybrid configuration of Ru/Ni-B-P || Pt/C being paired with the conventional benchmark electrode demonstrates a record low 2-E cell voltage of 2.40 V at 2000 mA cm-2 in 6 m KOH and excellent stability at high current of 1500 mA cm-2 under industrial operational condition.

Electrocatalytic Coenhancement of Bimetallic Polyphthalocyanine-Anchored Ru Nanoclusters Enabling Efficient Overall Water Splitting at Ampere-Level Current Densities.

Efficient electrocatalysts capable of operating continuously at industrial ampere-level current densities are crucial for large-scale applications of electrocatalytic water decomposition for hydrogen production. However, long-term industrial overall water splitting using a single electrocatalyst remains a major challenge. Here, bimetallic polyphthalocyanine (FeCoPPc)-anchored Ru nanoclusters, an innovative electrocatalyst comprising the hydrogen evolution reaction (HER) active Ru and the oxygen evolution reaction (OER) active FeCoPPc, engineered for efficient overall water splitting are demonstrated. By density functional theory calculations and systematic experiments, the electrocatalytic coenhancement effect resulting from unique charge redistribution, which synergistically boosts the HER activity of Ru and the OER activity of FeCoPPc by optimizing the adsorption energy of intermediates, is unveiled. As a result, even at a large current density of 2.0 A cm-2 , the catalyst exhibits low overpotentials of 220 and 308 mV, respectively, for HER and OER. It exhibits excellent stability, requiring only 1.88 V of cell voltage to achieve a current density of 2.0 A cm-2 in a 6.0 m KOH electrolyte at 70 °C, with a remarkable operational stability of over 100 h. This work provides a new electrocatalytic coenhancement strategy for the design and synthesis of electrocatalyst, paving the way for industrial-scale overall water splitting applications.

Dual Hydrogen Production System: Synergistic Effect of Ru and Ce Over Cu2O Nanotubes Drives Hydrogen Evolution and Formaldehyde Oxidation.

Water splitting for hydrogen production is limited by high cell voltage and low energy conversion efficiencies due to the slow kinetic process of the oxygen evolution reaction (OER). Here, an electrolytic system is constructed in which the cathode and anode co-release H2 at ultra-low input voltage using formaldehyde oxidation reaction (FOR) instead of OER. The prepared RuCe co-doped Cu2O nanotubes on copper foam (RuCe-Cu2O/CF) are used as electrode materials for the HER-FOR system. A current density of 0.8 A cm-2 is achieved at 0.55 V, and a stable hydrogen production process is realized at both the cathode and anode. Density functional theory (DFT) studies show that the synergistic effect of Ru and Ce drives: i) the d-band center of RuCe-Cu2O/CF away from the Fermi energy level; ii) the energy barrier for the C─H cracking of the H2C(OH)O* intermediate in FOR is lowered, which promotes the formation of H2 from H*, and iii) ΔGH* tends to 0 (-0.1 eV), optimizing the reaction kinetics of HER. This work provides a new design for an efficient catalyst for dual hydrogen production systems from water splitting.

Synergistic Effects of Amine Functional Groups and Enriched-Atomic-Iron Sites in Carbon Dots for Industrial-Current-Density CO2 Electroreduction.

Metal phthalocyanine molecules with Me-N4 centers have shown promise in electrocatalytic CO2 reduction (eCO2R) for CO generation. However, iron phthalocyanine (FePc) is an exception, exhibiting negligible eCO2R activity due to a higher CO2 to *COOH conversion barrier and stronger *CO binding energy. Here, amine functional groups onto atomic-Fe-rich carbon dots (Af-Fe-CDs) are introduced via a one-step solvothermal molecule fusion approach. Af-Fe-CDs feature well-defined Fe-N4 active sites and an impressive Fe loading (up to 8.5 wt%). The synergistic effect between Fe-N4 active centers and electron-donating amine functional groups in Af-Fe-CDs yielded outstanding CO2-to-CO conversion performance. At industrial-relevant current densities exceeding 400 mA cm-2 in a flow cell, Af-Fe-CDs achieved >92% selectivity, surpassing state-of-the-art CO2-to-CO electrocatalysts. The in situ electrochemical FTIR characterization combined with theoretical calculations elucidated that Fe-N4 integration with amine functional groups in Af-Fe-CDs significantly reduced energy barriers for *COOH intermediate formation and *CO desorption, enhancing eCO2R efficiency. The proposed synergistic effect offers a promising avenue for high-efficiency catalysts with elevated atomic-metal loadings.

Engineering Non-precious Trifunctional Cobalt-Based Electrocatalysts for Industrial Water Splitting and Ultra-High-Temperature Flexible Zinc-Air Battery.

Developing efficient, robust, and cost-effective trifunctional catalysts for the hydrogen evolution reaction (HER), oxygen evolution reaction (OER) and oxygen reduction reaction (ORR) at high current density and high temperature is crucial for water splitting at industry-level conditions and ultra-high-temperature Zinc-air battery (ZAB). Herein, cobalt nanoparticles well-integrated with nitrogen-doped porous carbon leaves (Co@NPCL) by direct annealing of core-shell bimetallic zeolite imidazolate frameworks is synthesized. Benefiting from the homogeneous distribution of metallic Co nanoparticles, the conductive porous carbon, and the doped N species, the as-fabricated Co@NPCL catalysts exhibit outstanding trifunctional performances with low overpotentials at 10 mA cm-2 for HER (87 mV) and OER (276 mV), long-lasting lifetime of over 2000 h, and a high half-wave potential of 0.86 V versus RHE for ORR. Meanwhile, the Co@NPCL catalyst can serve as both cathode and anode for water splitting at industrial conduction, and exhibit a stable cell voltage of 1.87 V to deliver a constant catalytic current of 500 mA cm-2 over 60 h. Moreover, the excellent trifunctional activity of Co@NPCL enables the flexible ZAB to operate efficiently at ultra-high temperature of 70 °C, delivering 162 mW cm-2 peaks power density and an impressive stability for 4500 min at 2 mA cm-2.

Finger millet (Eleusine coracana L.): from staple to superfood-a comprehensive review on nutritional, bioactive, industrial, and climate resilience potential.

MAIN CONCLUSION: This review discusses the Finger millet's rich nutritional profile, bioactive potential, and industrial applications, combined with its climate resilience, which make it a promising crop for enhancing food security and promoting sustainable agriculture. This review also highlights its significant potential to address malnutrition and mitigate climate change impacts. The emergence of Finger millet from "poor man's staple food" to "a nutrient rich cereal" has encouraged the need to explore this crop at a wider scale. It is a highly significant crop due to its rich nutritional and bioactive profile, diverse biological activities, and promising industrial applications, along with the high climate resilience. This comprehensive review evaluates its nutritional composition by comparing favorably with other cereals and millets and emphasizing its potential to address malnutrition and enhance food security. Furthermore, it explores the phytochemical/bioactive potential and strategies to enhance their bioavailability followed biological activities of Finger millet by highlighting its various health-promoting properties. The review also discusses industrial potential of finger millet including its role in nutraceutical and functional food production, as well as bioenergy generation. In addition, role of Finger millet as a climate-resilient crop; specifically, the available genetic resources and identification of genes and quantitative trait loci (QTLs) associated with major stress tolerance traits have also been discussed. By providing a comprehensive synthesis of existing knowledge, this study offers valuable insights for researchers, policymakers, and stakeholders engaged in efforts to promote sustainable agriculture, enhance food and nutrition security, and mitigate the impacts of climate change.

Custom integration of a magnetic-field monitoring system into a 32-channel MRI head coil.

PURPOSE: Customizing a Siemens 32-channel coil for use in a Philips 3T MRI system with incorporated magnetic field probes for collecting high-quality MRI and magnetic-field monitoring data concurrently. METHODS: The development process of the custom coil involved several (iterative) phases. Standard temporal SNR and B1 + data were collected with the 32-channel Siemens and for reference the 32-channel/8-channel Philips head coils before and after the custom coil was made compatible with the 3T Philips Achieva system, and magnetic field probes were installed into it along with ancillary electronics around it. Quality assurance tests were conducted in each of the build phases to ensure that the modifications did not affect MRI or field-monitoring data negatively. To test the finished custom coil, we collected high angular resolution diffusion imaging (HARDI) datasets on a spherical silicon oil phantom both with and without concurrent field monitoring and a 32-channel Philips coil without concurrent field monitoring, where the latter two served as reference scans to assess the improved performance of the custom coil with field monitoring. Similar HARDI-MRI data were also collected in vivo with the finished custom coil together with field monitoring data. RESULTS: The custom coil provided excellent temporal SNR especially at the edges where cortical gray matter is expected. When using concurrent field monitoring in HARDI acquisitions, the custom coil alleviated ghosting artifacts in phantom data and provided in vivo images with 1.4-mm isotropic resolution. CONCLUSION: The custom MRI coil with integrated magnetic-field monitoring probes provided improved imaging performance.

Flux optimization using multiple promoters in Halomonas bluephagenesis as a model chassis of the next generation industrial biotechnology.

Predictability and robustness are challenges for bioproduction because of the unstable intracellular synthetic activities. With the deeper understanding of the gene expression process, fine-tuning has become a meaningful tool for biosynthesis optimization. This study characterized several gene expression elements and constructed a multiple inducible system that responds to ten different small chemical inducers in halophile bacterium Halomonas bluephagenesis. Genome insertion of regulators was conducted for the purpose of gene cluster stabilization and regulatory plasmid simplification. Additionally, dynamic ranges of the multiple inducible systems were tuned by promoter sequence mutations to achieve diverse scopes for high-resolution gene expression control. The multiple inducible system was successfully employed to precisely control chromoprotein expression, lycopene and poly-3-hydroxybutyrate (PHB) biosynthesis, resulting in colorful bacterial pictures, optimized cell growth, lycopene and PHB accumulation. This study demonstrates a desirable approach for fine-tuning of rational and efficient gene expressions, displaying the significance for metabolic pathway optimization.

Multi-omic characterization of antibody-producing CHO cell lines elucidates metabolic reprogramming and nutrient uptake bottlenecks.

Characterizing the phenotypic diversity and metabolic capabilities of industrially relevant manufacturing cell lines is critical to bioprocess optimization and cell line development. Metabolic capabilities of production hosts limit nutrient and resource channeling into desired cellular processes and can have a profound impact on productivity. These limitations cannot be directly inferred from measured data such as spent media concentrations or transcriptomics. Here, we present an integrated multi-omic analysis pipeline combining exo-metabolomics, transcriptomics, and genome-scale metabolic network analysis and apply it to three antibody-producing Chinese Hamster Ovary cell lines to identify reprogramming features associated with high-producing clones and metabolic bottlenecks limiting product formation in an industrial bioprocess. Analysis of individual datatypes revealed a decreased nitrogenous byproduct secretion in high-producing clones and the topological changes in peripheral metabolic pathway expression associated with phase shifts. An integrated omics analysis in the context of the genome-scale metabolic model elucidated the differences in central metabolism and identified amino acid utilization bottlenecks limiting cell growth and antibody production that were not evident from exo-metabolomics or transcriptomics alone. Thus, we demonstrate the utility of a multi-omics characterization in providing an in-depth understanding of cellular metabolism, which is critical to efforts in cell engineering and bioprocess optimization.

Ectoine hyperproduction by engineered Halomonas bluephagenesis.

Ectoine, a crucial osmoprotectant for salt adaptation in halophiles, has gained growing interest in cosmetics and medical industries. However, its production remains challenged by stringent fermentation process in model microorganisms and low production level in its native producers. Here, we systematically engineered the native ectoine producer Halomonas bluephagenesis for ectoine production by overexpressing ectABC operon, increasing precursors availability, enhancing product transport system and optimizing its growth medium. The final engineered H. bluephagenesis produced 85 g/L ectoine in 52 h under open unsterile incubation in a 7 L bioreactor in the absence of plasmid, antibiotic or inducer. Furthermore, it was successfully demonstrated the feasibility of decoupling salt concentration with ectoine synthesis and co-production with bioplastic P(3HB-co-4HB) by the engineered H. bluephagenesis. The unsterile fermentation process and significantly increased ectoine titer indicate that H. bluephagenesis as the chassis of Next-Generation Industrial Biotechnology (NGIB), is promising for the biomanufacturing of not only intracellular bioplastic PHA but also small molecular compound such as ectoine.

A long-term growth stable Halomonas sp. deleted with multiple transposases guided by its metabolic network model Halo-ecGEM.

Microbial instability is a common problem during bio-production based on microbial hosts. Halomonas bluephagenesis has been developed as a chassis for next generation industrial biotechnology (NGIB) under open and unsterile conditions. However, the hidden genomic information and peculiar metabolism have significantly hampered its deep exploitation for cell-factory engineering. Based on the freshly completed genome sequence of H. bluephagenesis TD01, which reveals 1889 biological process-associated genes grouped into 84 GO-slim terms. An enzyme constrained genome-scale metabolic model Halo-ecGEM was constructed, which showed strong ability to simulate fed-batch fermentations. A visible salt-stress responsive landscape was achieved by combining GO-slim term enrichment and CVT-based omics profiling, demonstrating that cells deploy most of the protein resources by force to support the essential activity of translation and protein metabolism when exposed to salt stress. Under the guidance of Halo-ecGEM, eight transposases were deleted, leading to a significantly enhanced stability for its growth and bioproduction of various polyhydroxyalkanoates (PHA) including 3-hydroxybutyrate (3HB) homopolymer PHB, 3HB and 3-hydroxyvalerate (3HV) copolymer PHBV, as well as 3HB and 4-hydroxyvalerate (4HB) copolymer P34HB. This study sheds new light on the metabolic characteristics and stress-response landscape of H. bluephagenesis, achieving for the first time to construct a long-term growth stable chassis for industrial applications. For the first time, it was demonstrated that genome encoded transposons are the reason for microbial instability during growth in flasks and fermentors.

Saccharomyces cerevisiae strains performing similarly during fermentation of lignocellulosic hydrolysates show pronounced differences in transcriptional stress responses.

Improving our understanding of the transcriptional changes of Saccharomyces cerevisiae during fermentation of lignocellulosic hydrolysates is crucial for the creation of more efficient strains to be used in biorefineries. We performed RNA sequencing of a CEN.PK laboratory strain, two industrial strains (KE6-12 and Ethanol Red), and two wild-type isolates of the LBCM collection when cultivated anaerobically in wheat straw hydrolysate. Many of the differently expressed genes identified among the strains have previously been reported to be important for tolerance to lignocellulosic hydrolysates or inhibitors therein. Our study demonstrates that stress responses typically identified during aerobic conditions such as glutathione metabolism, osmotolerance, and detoxification processes also are important for anaerobic processes. Overall, the transcriptomic responses were largely strain dependent, and we focused our study on similarities and differences in the transcriptomes of the LBCM strains. The expression of sugar transporter-encoding genes was higher in LBCM31 compared with LBCM109 that showed high expression of genes involved in iron metabolism and genes promoting the accumulation of sphingolipids, phospholipids, and ergosterol. These results highlight different evolutionary adaptations enabling S. cerevisiae to strive in lignocellulosic hydrolysates and suggest novel gene targets for improving fermentation performance and robustness. IMPORTANCE: The need for sustainable alternatives to oil-based production of biochemicals and biofuels is undisputable. Saccharomyces cerevisiae is the most commonly used industrial fermentation workhorse. The fermentation of lignocellulosic hydrolysates, second-generation biomass unsuited for food and feed, is still hampered by lowered productivities as the raw material is inhibitory for the cells. In order to map the genetic responses of different S. cerevisiae strains, we performed RNA sequencing of a CEN.PK laboratory strain, two industrial strains (KE6-12 and Ethanol Red), and two wild-type isolates of the LBCM collection when cultivated anaerobically in wheat straw hydrolysate. While the response to inhibitors of S. cerevisiae has been studied earlier, this has in previous studies been done in aerobic conditions. The transcriptomic analysis highlights different evolutionary adaptations among the different S. cerevisiae strains and suggests novel gene targets for improving fermentation performance and robustness.

Pirfenidone ameliorates ANIT-induced cholestatic liver injury via modulation of FXR, NF-кB/TNF-α, and Wnt/GSK-3ß/ß-catenin signaling pathways.

Cholestasis is a hepatobiliary disorder characterized by the excessive accumulation of toxic bile acids in hepatocytes, leading to cholestatic liver injury (CLI) through multiple pathogenic inflammatory pathways. Currently, there are limited therapeutic options for the management of cholestasis and associated CLI; therefore, new options are urgently needed. Pirfenidone (PF), an oral bioavailable pyridone analog, is used for the treatment of idiopathic pulmonary fibrosis. PF has recently demonstrated diverse potential therapeutic activities against different pathologies. Accordingly, the present study adopted the α-naphthyl isothiocyanate (ANIT)-induced CLI model in mice to explore the potential protective impact of PF and investigate the underlying mechanisms of action. PF intervention markedly reduced the serum levels of ALT, AST, LDH, total bilirubin, and total bile acids, which was accompanied by a remarkable amelioration of histopathological lesions induced by ANIT. PF also protected the mice against ANIT-induced redox imbalance in the liver, represented by reduced MDA levels and elevated GSH and SOD activities. Mechanistically, PF inhibited ANIT-induced downregulated expressions of the farnesoid X receptor (FXR), as well as the bile salt export pump (BSEP) and the multidrug resistance-associated protein 2 (MRP2) bile acid efflux channels. PF further repressed ANIT-induced NF-κB activation and TNF-α and IL-6 production. These beneficial effects were associated with its ability to dose-dependently inhibit Wnt/GSK-3ß/ß-catenin/cyclin D1 signaling. Collectively, PF protects against ANIT-induced CLI in mice, demonstrating powerful antioxidant and anti-inflammatory activities as well as an ability to oppose BA homeostasis disorder. These protective effects are primarily mediated by modulating the interplay between FXR, NF-κB/TNF-α/IL-6, and Wnt/ß-catenin signaling pathways.