Riboflavin Research Articles

Unlocking multi-mode sensing potential: Phosphorus-doped graphitic carbon nitride quantum dots for Ag+, ciprofloxacin, and riboflavin analysis in environment and food matrices

The simultaneous detection of multiple analytes through a single fluorescence sensor is highly attractive. In this study, phosphorus-doped graphitic carbon nitride quantum dots (P-CNQDs) were developed, achieving multi-mode sensing through three distinct response mechanisms. The preparation involved using melamine as the carbon and nitrogen source and ammonium dihydrogen phosphate as the phosphorus source. Uniform and narrowly distributed P-CNQDs were successfully synthesized through chemical oxidation and hydrothermal methods, with an average size of 2.4 nm. These unique P-CNQDs exhibited fluorescence quenching through photo-induced electron transfer (PET) in response to Ag+. Additionally, the formation of hydrogen bonds and coordination interactions between P-CNQDs-Ag+ and ciprofloxacin (CIP) led to a pronounced fluorescence response to CIP by the chelation enhanced fluorescence (CHEF) mechanism. Furthermore, leveraging the principle of fluorescence resonance energy transfer (FRET), P-CNQDs-CIP served as a ratio fluorescence sensor for riboflavin (RF), enabling ultra-sensitive detection of RF. The combination of PET, CHEF, and FRET response mechanisms successfully facilitated multi-mode sensing for Ag+, CIP, and RF. The detection ranges were 0.05–100 μM, 0.002–2 μM, and 0.05–60 μM, with corresponding lowest detection limits of 17.1 nM, 1.1 nM, and 29.2 nM, respectively. This versatile sensor has been effectively applied to real samples, including the detection of river water and vitamin B2 tablets.

Comparative analysis of the growth differences between hybrid Ningdu Yellow chickens and their parentals

Although the local high quality chicken breed in China has excellent flavor, its growth rate is inferior to that of foreign breeds. To improve the growth rate of local chicken breeds, it is crucial to study the mechanism of chicken muscle development. Herein, Ningdu Yellow chicken was used as the research object, and a new hybrid breed (W) was obtained by crossing the G, H and D lines, which combined the excellent physiological characteristics of its parents. Combined analysis of Ningdu Yellow chickens and their parents was carried out. Chickens from 105-day-old lines (W, G, H) were selected, and breast meat and serum were extracted for transcriptome sequencing and metabolome determination to study their growth differences. The live weight, carcass weight, half-eviscerated weight, eviscerated weight, and breast muscle weight of W were significantly higher than those of G and H. Differential expression analysis identified 1700 differentially expressed genes (DEG), and gene ontology and kyoto encyclopedia of genes and genomes (KEGG) analyses identified 33 and 1 pathways related to growth and development and steroid biosynthesis, respectively. Next, pairwise analysis identified 57 KEGG pathways, among which the MAPK signaling, steroid hormone biosynthesis, tight junction, and PPAR signaling pathways were involved in growth and development. Cluster analysis found that genes highly expressed in the W group were associated with regulation of the actin cytoskeleton, riboflavin metabolism, steroid biosynthesis, and glycerophospholipid metabolism. The top 2 clusters obtained by protein-protein interaction analysis were important for the growth and development of chickens. Finally, the metabolomic analysis found key differentially accumulated metabolites (DAM) that might be account for the growth differences. Further integrated analysis identified key DEGs and DAMs that might be responsible for the observed growth differences. This study identified genes governing growth traits in Ningdu Yellow chickens, laying a theoretical foundation for the development of chicken breeding, the utilization of hybrid supporting lines, and promotion of the Chinese chicken industry.

Key genomes, transcriptomes, proteins, and metabolic factors involved in the detoxification/tolerance of TNT and its intermediates by bacteria in anaerobic/aerobic environments

Novel microbial strains capable of efficient degradation of TNT and typical intermediates (2-ADNT and 4-ADNT) in aerobic/anaerobic environment were screened and isolated from ammunition-contaminated sites. The key genomes, transcriptomes, proteins, and metabolic factors for microbial detoxification/tolerance to pollutants in anaerobic and aerobic environments were analyzed for the first time. The bacterial genome, which is rich in metabolism and environmental information-processing functional genes, provides transcriptional and translational-related proteins for detoxifying/tolerating pollutants. At the transcriptional level, bacteria significantly expressed genes related to inositol phosphate metabolism for regulating membrane transport, maintaining the cytoskeleton, and signal transduction. At the protein level, genes involved in antioxidation, fat metabolism, sugar synthesis/degradation, and pyruvate metabolism were significantly expressed. At the metabolic level, riboflavin metabolism, which regulates membrane integrity, protects against oxidative stress, and maintains the sugar–protein–fat balance, showed significant responses. Bacteria simultaneously regulate amino acid metabolism, carbohydrate metabolism, and N/P/S cycles to maintain homeostatic cellular energy supplies. The key pathway for pollutant degradation in bacteria is nitrotoluene degradation. The molecular mechanism of bacterial tolerance to pollutants involves the regulation of oxidative phosphorylation and basic cycle pathways to maintain gene transcription, protein translation, and metabolic cycles.

In-situ Growth of CuO Nanoflakes on Graphitic Carbon Nitride Sheets: An Electro‐active Interface for Electrocatalytic Oxidation and Detection of Riboflavin in Food and Nutritional Supplements

The altered riboflavin (RF) level is considered a biomarker for early diagnosis of human ailments and also serves as an internal marker for tracing food quality and adulteration. This necessitates the need for a sensitive, selective, and affordable method for RF estimation in real samples. RF, a biological chelating ligand, is known to have an affinity for complex formation with Cu(II) by coordinating through the electron-rich nitrogen and oxygen atoms on its structure. Motivated by this, CuO nanoflakes grown over graphitic carbon nitride (gCN) support is presented as an electroactive interface (gCN.CuNF|GCE) for electrochemical RF detection. A systematic analysis of the tailored electroactive interface is presented using HR-SEM, XRD, FT-IR, and XPS. The electrochemical analysis reveals the electro-oxidation of RF at gCN.CuNF|GCE with a 4.6 times higher current and a ∼13 mV potential shift, outlining the electro-catalytic activity of the composite material towards RF. The developed sensor exhibited a discernible peak even for 25 nM RF, showcasing an LOD of 6 nM. Furthermore, excellent selectivity for RF was observed even with potential interfering species, including cyanocobalamin. Besides detectability at the nanomolar range, excellent performance was verified for repeatability and RF analysis in food and pharmaceutical samples.

Varicella Zoster Virus disrupts MAIT cell polyfunctional effector responses.

Mucosal-associated invariant T (MAIT) cells are unconventional T cells that respond to riboflavin biosynthesis and cytokines through TCR-dependent and -independent pathways, respectively. MAIT cell activation plays an immunoprotective role against several pathogens, however the functional capacity of MAIT cells following direct infection or exposure to infectious agents remains poorly defined. We investigated the impact of Varicella Zoster Virus (VZV) on blood-derived MAIT cells and report virus-mediated impairment of activation, cytokine production, and altered transcription factor expression by VZV infected (antigen+) and VZV exposed (antigen-) MAIT cells in response to TCR-dependent and -independent stimulation. Furthermore, we reveal that suppression of VZV exposed (antigen-) MAIT cells is not mediated by a soluble factor from neighbouring VZV infected (antigen+) MAIT cells. Finally, we demonstrate that VZV impairs the cytolytic potential of MAIT cells in response to riboflavin synthesising bacteria. In summary, we report a virus-mediated immune-evasion strategy that disarms MAIT cell responses.

Identification of differential gene expression related to reproduction in the sporophytes of Saccharina japonica.

Saccharina japonica, a significant brown macroalga in the Pacific Ocean, serves as a food source and industrial material. In aquaculture, collecting mature sporophytes for seedling production is essential but challenging due to environmental changes. In this study, transcriptomic analysis of vegetative and sorus tissues was done to identify differentially expressed genes (DEGs) and enhance our understanding of sorus formation regulation in S. japonica. KEGG pathway and Gene Otology (GO) analysis revealed that upregulated DEGs were involved in folate biosynthesis, riboflavin metabolism, and amino acid biosynthesis. In addition, the upregulation of genes associated with cell wall remodeling, such as mannuronan C-5-epimerases, vanadium-dependent haloperoxidases, and NADPH oxidase, was observed in sorus parts. Meanwhile, downregulated DEGs in sorus portions included genes related to chloroplast function. These findings will help us understand the regulatory mechanisms behind sorus formation in S. japonica and extracellular matrix remodeling in brown algae.

Riboflavin-meditated interspecies electron transfer in a H2-based denitrifying biofilm

H2-based membrane biofilm reactors (MBfR) are effective for nitrogen removal, yet the interactions of microorganisms within the MBfR biofilms have received limited attention. In this study, we explored a flavin-mediated electron transport mechanism occurring between hydrogenotrophic microorganisms and non‑hydrogenotrophic denitrifying microorganisms in H2-based MBfR biofilms. Liquid chromatography-tandem mass spectrometry (LC-MS/MS) and electrochemical analysis confirmed riboflavin act as the sole potential redox mediator present in the system. Batch experiments demonstrated that riboflavin accelerates the denitrification process and that microorganisms can use reduced flavins as the sole electron donor to reduce nitrate. Metagenomic and metatranscriptomic analyses revealed increased expression of genes related to riboflavin biosynthesis and transport in hydrogenotrophic microorganisms, indicating endogenous riboflavin secretion into the medium. Our findings reveal the potential existence of a novel flavin-mediated electron transfer mechanism in the H₂-based denitrification process, enriching the understanding of interspecies interactions within MBfR biofilms. This study also provides a theoretical basis for efficient nitrogen removal through regulating interspecies electron transfer.

Enhancing gut microbiota and microbial function with inulin supplementation in children with obesity.

Gut dysbiosis that resulted from the alteration between host-microbe interaction might worsen obesity-induced systemic inflammation. Gut microbiota manipulation by supplementation of prebiotic inulin may reverse metabolic abnormalities and improve obesity. This study aimed to determine whether inulin supplementation improved intestinal microbiota and microbial functional pathways in children with obesity. Children with obesity whose BMI above median + 2SDs were recruited to a randomized, double-blinded placebo-controlled study. The participants aged 7-15 years were assigned to inulin supplement extracted from Thai Jerusalem artichoke (intervention), maltodextrin (placebo), and dietary fiber advice groups. All participants received similar monthly conventional advice and follow-up for 6 months. Fecal samples were collected for gut microbiome analysis using 16S rRNA sequencing. Phylogenetic Investigation of Communities by Reconstruction of Unobserved States was performed to infer microbial functional pathways. One hundred and forty-three children with available taxonomic and functional pathway abundance profiles were evaluated. A significant increase in alpha-diversity was observed in the inulin group. Inulin supplementation substantially enhanced Bifidobacterium, Blautia, Megasphaera, and several butyrate-producing bacteria, including Agathobacter, Eubacterium coprostanoligenes, and Subdoligranulum, compared to the other groups. The inulin group showed a significant difference in functional pathways of proteasome and riboflavin metabolism. These changes correlated with clinical and metabolic outcomes exclusively in the inulin group. Inulin supplementation significantly promoted gut bacterial diversity and improved gut microbiota dysbiosis in children with obesity. The modulation of functional pathways by inulin suggests its potential to establish beneficial interactions between the gut microbiota and host physiology. Inulin supplementation could be a strategic treatment to restore the balance of intestinal microbiota and regulate their functions in childhood obesity.

AtDREB2G is involved in the regulation of riboflavin biosynthesis in response to low-temperature stress and abscisic acid treatment in Arabidopsis thaliana

Riboflavin (RF) serves as a precursor to flavin mononucleotide and flavin adenine dinucleotide, which are crucial cofactors in various metabolic processes. Strict regulation of cellular flavin homeostasis is imperative, yet information regarding the factors governing this regulation remains largely elusive. In this study, we first examined the impact of external flavin treatment on the Arabidopsis transcriptome to identify novel regulators of cellular flavin levels. Our analysis revealed alterations in the expression of 49 putative transcription factors. Subsequent reverse genetic screening highlighted a member of the dehydration-responsive element binding (DREB) family, AtDREB2G, as a potential regulator of cellular flavin levels. Knockout mutants of AtDREB2G (dreb2g) exhibited reduced flavin levels and decreased expression of RF biosynthetic genes compared to wild-type plants. Conversely, conditional overexpression of AtDREB2G led to an increase in the expression of RF biosynthetic genes and elevated flavin levels. In wild-type plants, exposure to low temperatures and abscisic acid treatment stimulated enhanced flavin levels and upregulated the expression of RF biosynthetic genes, concomitant with the induction of AtDREB2G. Notably, these responses were significantly attenuated in dreb2g mutants. Our findings establish AtDREB2G is involved in the positive regulation of flavin biosynthesis in Arabidopsis, particularly under conditions of low temperature and abscisic acid treatment.

Integrated network pharmacology and metabolomics to reveal the mechanism of Pinellia ternata inhibiting non-small cell lung cancer cells

Lung cancer is a malignant tumor with highly heterogeneous characteristics. A classic Chinese medicine, Pinellia ternata (PT), was shown to exert therapeutic effects on lung cancer cells. However, its chemical and pharmacological profiles are not yet understood. In the present study, we aimed to reveal the mechanism of PT in treating lung cancer cells through metabolomics and network pharmacology. Metabolomic analysis of two strains of lung cancer cells treated with Pinellia ternata extracts (PTE) was used to identify differentially abundant metabolites, and the metabolic pathways associated with the DEGs were identified by MetaboAnalyst. Then, network pharmacology was applied to identify potential targets against PTE-induced lung cancer cells. The integrated network of metabolomics and network pharmacology was constructed based on Cytoscape. PTE obviously inhibited the proliferation, migration and invasion of A549 and NCI-H460 cells. The results of the cellular metabolomics analysis showed that 30 metabolites were differentially expressed in the lung cancer cells of the experimental and control groups. Through pathway enrichment analysis, 5 metabolites were found to be involved in purine metabolism, riboflavin metabolism and the pentose phosphate pathway, including D-ribose 5-phosphate, xanthosine, 5-amino-4-imidazolecarboxyamide, flavin mononucleotide (FMN) and flavin adenine dinucleotide (FAD). Combined with network pharmacology, 11 bioactive compounds were found in PT, and networks of bioactive compound–target gene–metabolic enzyme–metabolite interactions were constructed. In conclusion, this study revealed the complicated mechanisms of PT against lung cancer. Our work provides a novel paradigm for identifying the potential mechanisms underlying the pharmacological effects of natural compounds.

Effects of different energy levels in low-protein diet on liver lipid metabolism in the late-phase laying hens through the gut-liver axis

BackgroundThe energy/protein imbalance in a low-protein diet induces lipid metabolism disorders in late-phase laying hens. Reducing energy levels in the low-protein diet to adjust the energy-to-protein ratio may improve fat deposition, but this also decreases the laying performance of hens. This study investigated the mechanism by which different energy levels in the low-protein diet influences liver lipid metabolism in late-phase laying hens through the enterohepatic axis to guide feed optimization and nutrition strategies. A total of 288 laying hens were randomly allocated to the normal-energy and normal-protein diet group (positive control: CK) or 1 of 3 groups: low-energy and low-protein diet (LL), normal-energy and low-protein diet (NL), and high-energy and low-protein diet (HL) groups. The energy-to-protein ratios of the CK, LL, NL, and HL diets were 0.67, 0.74, 0.77, and 0.80, respectively.ResultsCompared with the CK group, egg quality deteriorated with increasing energy intake in late-phase laying hens fed low-protein diet. Hens fed LL, NL, and HL diets had significantly higher triglyceride, total cholesterol, acetyl-CoA carboxylase, and fatty acid synthase levels, but significantly lower hepatic lipase levels compared with the CK group. Liver transcriptome sequencing revealed that genes involved in fatty acid beta-oxidation (ACOX1, HADHA, EHHADH, and ACAA1) were downregulated, whereas genes related to fatty acid synthesis (SCD, FASN, and ACACA) were upregulated in LL group compared with the CK group. Comparison of the cecal microbiome showed that in hens fed an LL diet, Lactobacillus and Desulfovibrio were enriched, whereas riboflavin metabolism was suppressed. Cecal metabolites that were most significantly affected by the LL diet included several vitamins, such as riboflavin (vitamin B2), pantethine (vitamin B5 derivative), pyridoxine (vitamin B6), and 4-pyridoxic acid.ConclusionA lipid metabolism disorder due to deficiencies of vitamin B2 and pantethine originating from the metabolism of the cecal microbiome may be the underlying reason for fat accumulation in the liver of late-phase laying hens fed an LL diet. Based on the present study, we propose that targeting vitamin B2 and pantethine (vitamin B5 derivative) might be an effective strategy for improving lipid metabolism in late-phase laying hens fed a low-protein diet.

Structure of Aquifex aeolicus lumazine synthase by cryo-electron microscopy to 1.42 Å resolution.

Single-particle cryo-electron microscopy (cryo-EM) has become an essential structural determination technique with recent hardware developments making it possible to reach atomic resolution, at which individual atoms, including hydrogen atoms, can be resolved. In this study, we used the enzyme involved in the penultimate step of riboflavin biosynthesis as a test specimen to benchmark a recently installed microscope and determine if other protein complexes could reach a resolution of 1.5 Å or better, which so far has only been achieved for the iron carrier ferritin. Using state-of-the-art microscope and detector hardware as well as the latest software techniques to overcome microscope and sample limitations, a 1.42 Å map of Aquifex aeolicus lumazine synthase (AaLS) was obtained from a 48 h microscope session. In addition to water molecules and ligands involved in the function of AaLS, we can observe positive density for ∼50% of the hydrogen atoms. A small improvement in the resolution was achieved by Ewald sphere correction which was expected to limit the resolution to ∼1.5 Å for a molecule of this diameter. Our study confirms that other protein complexes can be solved to near-atomic resolution. Future improvements in specimen preparation and protein complex stabilization may allow more flexible macromolecules to reach this level of resolution and should become a priority of study in the field.

Fluoride induces immunotoxicity by regulating riboflavin transport and metabolism partly through IL-17A in the spleen

The impairment of the immune system by fluoride is a public health concern worldwide, yet the underlying mechanism is unclear. Both riboflavin and IL-17A are closely related to immune function and regulate the testicular toxicity of fluoride. However, whether riboflavin or IL-17A is involved in fluoride-induced immunotoxicity is unknown. Here, we first established a male ICR mouse model by treating mice with sodium fluoride (NaF) (100 mg/L) via the drinking water for 91 days. The results showed that fluoride increased the expression of the proinflammatory factors IL-1β and IL-17A, which led to splenic inflammation and morphological injury. Moreover, the expression levels of the riboflavin transporters SLC52A2 and SLC52A3; the transformation-related enzymes RFK and FLAD1; and the key mitochondrial functional determinants SDH, COX, and ATP in the spleen were measured via real-time PCR, Western blotting, and ELISA. The results revealed that fluoride disrupted riboflavin transport, transformation, metabolism, and mitochondrial function. Furthermore, wild-type (WT) and IL-17A knockout (IL-17A-/-) C57BL/6 J male mice of the same age were treated with NaF (24 mg/kg·bw, equivalent to 100 mg/L) and/or riboflavin sodium phosphate (5 mg/kg·bw) via gavage for 91 days. Similar parameters were evaluated as above. The results confirmed that fluoride increased riboflavin metabolism through RFK but not through FLAD1. Fluoride also affected mitochondrial function and activated neutrophils (marked with Ly6g) and macrophages (marked with CD68) in the spleen. Interestingly, IL-17A partly mediated fluoride-induced riboflavin metabolism disorder and immunotoxicity in the spleen. This work not only reveals a novel toxic mechanism for fluoride but also provides new clues for exploring the physiological function of riboflavin and for diagnosing and treating the toxic effects of fluoride in the environment.

Fe0-to-microbe electron transfer corrosion of EH36 steel by planktonic cells of Desulfovibrio vulgaris mediated by riboflavin

This study investigated the Fe0-to-microbe extracellular electron transfer (EET) corrosion of EH36 steel by planktonic Desulfovibrio vulgaris cells using Tetrakis hydroxymethyl phosphonium sulfate (THPS), peptide E6, and riboflavin (RF) as tools. 60 min after 20 ppm THPS & E6 injection and 50 ppm THPS injection, there was no difference in sessile cell counts, but planktonic cell counts were different. The subsequent injection of 20 ppm RF accelerated D. vulgaris MIC in varying degrees within minutes. This was attributed to RF-mediated EET of planktonic cells to accelerate corrosion, which can be further confirmed by the elevated expression levels of cytochrome c.

Influence of riboflavin on the corrosion of X80 pipeline steel by Sulfate reducing bacteria

The sulfate reducing bacteria(SRB) is commonly attached to the surface of buried pipeline steel, and the electron shuttle in the corrosion medium can promote the release of electrons from iron oxidation through the bacterial cell wall into the cytoplasm to accelerate the corrosion of anode iron. This study investigated the impact of riboflavin (RF) as an endogenous electron shuttle on the corrosion behavior of X80 pipeline steel in SRB system. The findings indicated that while the type of corrosion products remains unchanged in samples under SRB+10 mg l−1 RF system, there was an expansion in both area and depth of corrosion pits on the sample surface, resulting in a corrosion loss rate approximately 3 times higher than that observed in SRB system. Furthermore, the polarization resistance (Rp) value of the sample in SRB system is about 2 ∼ 5 times that of the sample in SRB+10 mg l−1 RF system. Additionally, the corrosion current density of X80 pipeline steel samples soaked in SRB and SRB +10 mg l−1 RF system for 14 days is 9.31 × 10-6 A·cm−2 and 1.28 × 10−5 A·cm−2, and the addition of 10 mg l−1 RF increases the corrosion current density of SRB system by about 37.49%. These results indicated that the reaction resistance of SRB-induced MIC in X80 pipeline steel was significantly reduced due to the presence of RF.

One-step green synthesis of platinum mesoporous nanoparticles by riboflavin for light activated antitumoral therapy

Photodynamic therapy (PDT) has been established as one of the most promising novel cancer therapies with fewer side-effects and enhanced efficacy compared to the currently available conventional treatments. However, its application has been hindered by the limitations that photosensitizers (PS) have. The combination of PS with metallic nanoparticles like platinum nanoparticles (PtNPs), can help to overcome these intrinsic drawbacks. In this work, the combination of PtNPs and the natural photosensitizer riboflavin (RF) is proposed. PtNPs are synthesized using RF (Pt@RF) as reducing and stabilizing agent in a one-step method, obtaining nanoparticles with mesoporous structure for UV triggered PDT. In view of possible future UV irradiation treatments, the degradation products of RF, ribitol (RB) and lumichrome (LC), this last being a photosensitizing byproduct, are also employed for the synthesis of porous PtNPs, obtaining Pt@LC and Pt@RB. When administered in vitro to lung cancer cells, all the samples elicit a strong decrease of cell viability and a decrease of intracellular ATP levels. The antitumoral effect of both Pt@RF and Pt@LC is triggered by UV-A irradiation. This antitumoral activity is caused by the induction of oxidative stress, shown in our study by the decrease in intracellular glutathione and increased expression of antioxidant enzymes.

Rats exposed to Alternaria toxins in vivo exhibit altered liver activity highlighted by disruptions in riboflavin and acylcarnitine metabolism

Natural toxins produced by Alternaria fungi include the mycotoxins alternariol, tenuazonic acid and altertoxins I and II. Several of these toxins have shown high toxicity even at low levels including genotoxic, mutagenic, and estrogenic effects. However, the metabolic effects of toxin exposure from Alternaria are understudied, especially in the liver as a key target. To gain insight into the impact of Alternaria toxin exposure on the liver metabolome, rats (n = 21) were exposed to either (1) a complex culture extract with defined toxin profiles from Alternaria alternata (50 mg/kg body weight), (2) the isolated, highly genotoxic altertoxin-II (ATX-II) (0.7 mg/kg of body weight) or (3) a solvent control. The complex mixture contained a spectrum of Alternaria toxins including a controlled dose of ATX-II, matching the concentration of the isolated ATX-II. Liver samples were collected after 24 h and analyzed via liquid chromatography–high-resolution mass spectrometry (LC-HRMS). Authentic reference standards (> 100) were used to identify endogenous metabolites and exogenous compounds from the administered exposures in tandem with SWATH-acquired MS/MS data which was used for non-targeted analysis/screening. Screening for metabolites produced by Alternaria revealed several compounds solely isolated in the liver of rats exposed to the complex culture, confirming results from a previously performed targeted biomonitoring study. This included the altersetin and altercrasin A that were tentatively identified. An untargeted metabolomics analysis found upregulation of acylcarnitines in rats receiving the complex Alternaria extract as well as downregulation of riboflavin in rats exposed to both ATX-II and the complex mixture. Taken together, this work provides a mechanistic view of Alternari toxin exposure and new suspect screening insights into hardly characterized Alternaria toxins.

The response mechanism analysis of HMX1 knockout strain to levulinic acid in Saccharomyces cerevisiae.

Levulinic acid, a hydrolysis product of lignocellulose, can be metabolized into important compounds in the field of medicine and pesticides by engineered strains of Saccharomyces cerevisiae. Levulinic acid, as an intermediate product widely found in the conversion process of lignocellulosic biomass, has multiple applications. However, its toxicity to Saccharomyces cerevisiae reduces its conversion efficiency, so screening Saccharomyces cerevisiae genes that can tolerate levulinic acid becomes the key. By creating a whole-genome knockout library and bioinformatics analysis, this study used the phenotypic characteristics of cells as the basis for screening and found the HMX1 gene that is highly sensitive to levulinic acid in the oxidative stress pathway. After knocking out HMX1 and treating with levulinic acid, the omics data of the strain revealed that multiple affected pathways, especially the expression of 14 genes related to the cell wall and membrane system, were significantly downregulated. The levels of acetyl-CoA and riboflavin decreased by 1.02-fold and 1.44-fold, respectively, while the content of pantothenic acid increased. These findings indicate that the cell wall-membrane system, as well as the metabolism of acetyl-CoA and riboflavin, are important in improving the resistance of Saccharomyces cerevisiae to levulinic acid. They provide theoretical support for enhancing the tolerance of microorganisms to levulinic acid, which is significant for optimizing the conversion process of lignocellulosic biomass to levulinic acid.

Modulation of riboflavin biosynthesis and utilization in mycobacteria.

The pathway for biosynthesis and utilization of riboflavin, precursor of the essential coenzymes, FMN and FAD, is of particular interest in the flavin-rich pathogen, Mycobacterium tuberculosis (Mtb), for two important reasons: (i) the pathway includes potential tuberculosis (TB) drug targets and (ii) intermediates from the riboflavin biosynthesis pathway provide ligands for mucosal-associated invariant T (MAIT) cells, which have been implicated in TB pathogenesis. However, the riboflavin pathway is poorly understood in mycobacteria, which lack canonical mechanisms to transport this vitamin and to regulate flavin coenzyme homeostasis. By conditionally disrupting each step of the pathway and assessing the impact on mycobacterial viability and on the levels of the pathway proteins as well as riboflavin, our work provides genetic validation of the riboflavin pathway as a target for TB drug discovery and offers a resource for further exploring the association between riboflavin biosynthesis, MAIT cell activation, and TB infection and disease.

Photodegradation of the novel herbicide pyraclonil in aqueous solution: Kinetics, identification of photoproducts, mechanism, and toxicity assessment

Pyraclonil is a new type of pyrazole herbicide, whose photochemical fate in aqueous solution has not been reported yet. In this study, effects on the photolysis rate such as light source, pH, NO3−, Fe3+, fulvic acid (FA) and riboflavin (RF) were investigated. Pyraclonil photodegraded in pure water under both UV and simulated sunlight with half-lives of 32.29 min and 42.52 h, respectively. Under UV, the degradation rate of pyraclonil in pH 4 solution (0.0299 ± 0.0033 min−1) was about twice higher than that in pH 9 (0.0160 ± 0.0063 min−1). Under simulated sunlight, low concentration (0.1–1 mg/L) of FA, NO3−, Fe3+ and RF noticeably promoted the photodegradation of pyraclonil. Then, with the combination of experimental UPLC-Q-TOF/MS and computational calculation of density functional theory (DFT), fourteen transformation products (TPs) of pyraclonil were identified with possible mechanism of C–N bond cleavage, photorearrangement, demethylation, hydroxylation and oxidation. Additionally, acute toxicity assessment was conducted through ECOSAR prediction and laboratory bioassays. The prediction results indicated that toxicity of TP157 to daphnid and green algae was 1.3 and 1.4 times higher than that of the parent, respectively. The bioassay results indicated that toxicities of TP157 and TP263 to C. vulgaris were about 1.6 and 5.9 times higher than that of the parent, respectively. The results provided a reference for elucidating the potential hazards of pyraclonil to non-target organisms and promoting its rational use.