Biotransformation Of Metabolites Research Articles

An effective LC-MS method for the simultaneous determination of a potential anti-rheumatoid arthritis drug, carboxyamidotriazole, and its major metabolite in rat plasma.

Carboxyamidotriazole (CAI) was previously recognized as a well-tolerated anticancer drug. It has also demonstrated significant anti-inflammatory effects in various cell and animal model experiments, prompting its investigation as a potential treatment for rheumatoid arthritis. In this study, the potential biotransformation metabolites of CAI were identified both in vitro and in vivo. A sensitive, specific, and accurate LC-MS method was developed for the quantitative analysis of CAI and its major metabolite, CAI-OH, in rat plasma. CAI, CAI-OH, and telmisartan (used as an internal standard) were separated using a Zorbax SB C18 column. The mobile phase consisted of water (phase A, containing 0.1% formic acid) and acetonitrile (phase B, containing 0.1% formic acid) at a flow rate of 0.2 mL/min. The analytes were examined using a high-resolution mass spectrometer, with detected mass-to-charge ratios of m/z 424.01293 for CAI, m/z 440.00785 for CAI-OH, and m/z 515.24415 for telmisartan. Good linearity was observed within the range of 10-5000 ng/mL. Both inter- and intra-batch precision (relative standard deviation, %) were below 6%, and the accuracy ranged from 94.9% to 106.1%. The analytes remained stable throughout the entire experimental period. This method was successfully applied in a pharmacokinetic study of CAI following oral administration in rats.

In Vitro and In Vivo Biotransformation Profiling of 6PPD-Quinone toward Their Detection in Human Urine.

The antioxidant N-(1,3-Dimethylbutyl)-N'-phenyl-p-phenylenediamine (6PPD) and its oxidized quinone product 6PPD-quinone (6PPD-Q) in rubber have attracted attention due to the ecological risk that they pose. Both 6PPD and 6PPD-Q have been detected in various environments that humans cohabit. However, to date, a clear understanding of the biotransformation of 6PPD-Q and a potential biomarker for exposure in humans are lacking. To address this issue, this study presents a comprehensive analysis of the extensive biotransformation of 6PPD-Q across species, encompassing both in vitro and in vivo models. We have tentatively identified 17 biotransformation metabolites in vitro, 15 in mice in vivo, and confirmed the presence of two metabolites in human urine samples. Interestingly, different biotransformation patterns were observed across species. Through semiquantitative analysis based on peak areas, we found that almost all 6PPD-Q underwent biotransformation within 24 h of exposure in mice, primarily via hydroxylation and subsequent glucuronidation. This suggests a rapid metabolic processing of 6PPD-Q in mammals, underscoring the importance of identifying effective biomarkers for exposure. Notably, monohydroxy 6PPD-Q and 6PPD-Q-O-glucuronide were consistently the most predominant metabolites across our studies, highlighting monohydroxy 6PPD-Q as a potential key biomarker for epidemiological research. These findings represent the first comprehensive data set on 6PPD-Q biotransformation in mammalian systems, offering insights into the metabolic pathways involved and possible exposure biomarkers.

Identification and characterization of chemical constituents in Mahuang Guizhi Decoction and their metabolites in rat plasma and brain by UPLC-Q-TOF/MS

ObjectiveMahuang Guizhi Decoction (MGD), an essential herbal pair in traditional Chinese medicine, is able to release cold, fever and asthma, mainly containing alkaloids, flavonoids, phenylpropanoids and amino acids. However, the absorption and distribution of these four category compounds in vivo still remained unclearly. MethodsIn our research, we utilized UPLC-Q-TOF-MS technique to identify the constituents within MGD, as well as the prototypes of MGD and their metabolites absorbed in plasma and brain. We further profiled the drug-time curve of prototypes and metabolites of MGD both in plasma and brain. ResultsOur results showed that 105 constituents were characterized in MGD. Thirty of them could be absorbed into blood, and ten of them could be distributed into brain. We also discovered eight new bio-transformed metabolites in blood, and a half of which could pass through the blood–brain barrier. In addition, all components detected in vivo could be absorbed and distributed immediately. ConclusionThese findings provide an approachable method to analyze the potential bio-active compounds in MGD and their in vivo behaviors, which could promote the efficacious material basis study of MGD and the security of clinical utilization.

Discovery of a series of portimine-A fatty acid esters in mussels

Vulcanodinium rugosum is a benthic dinoflagellate known for producing pinnatoxins, pteriatoxins, portimines and kabirimine. In this study, we aimed to identify unknown analogs of these emerging toxins in mussels collected in the Ingril lagoon, France. First, untargeted data acquisitions were conducted by means of liquid chromatography coupled to hybrid quadrupole-orbitrap mass spectrometry. Data processing involved a molecular networking approach, and a workflow dedicated to the identification of biotransformed metabolites. Additionally, targeted analyses by liquid chromatography coupled to triple quadrupole mass spectrometry were also implemented to further investigate and confirm the identification of new compounds. For the first time, a series of 13-O-acyl esters of portimine-A (n = 13) were identified, with fatty acid chains ranging between C12:0 and C22:6. The profile was dominated by the palmitic acid conjugation. This discovery was supported by fractionation experiments combined with the implementation of a hydrolysis reaction, providing further evidence of the metabolite identities. Furthermore, several analogs were semi-synthesized, definitively confirming the discovery of these metabolization products. A new analog of pinnatoxin, with a molecular formula of C42H65NO9, was also identified across the year 2018, with the highest concentration observed in August (4.5 μg/kg). The MS/MS data collected for this compound exhibited strong structural similarities with PnTX-A and PnTX-G, likely indicating a substituent C2H5O2 in the side chain at C33. The discovery of these new analogs will contribute to deeper knowledge of the chemodiversity of toxins produced by V. rugosum or resulting from shellfish metabolism, thereby improving our ability to characterize the risks associated with these emerging toxins.

Biodegradation of furfuryl alcohol by indigenous Bacillus species of industrial effluent-contaminated sites: estimation, biokinetics and toxicity assessment of bio-transformed metabolites.

Furfuryl alcohol (FA) and other furanic compounds have garnered considerable attention in the quest for sustainable alternatives. FA-based resins have been used in various sectors that entail the release of FA into the environment. Hence, to ensure sustainability in this scenario, devising a dependable approach to its degradation is imperative. Given the crucial role of bacterial strains in the biodegradation of various organic pollutants, this study investigates the microbial degradation of FA, using bacterial strains isolated from sites that are constantly exposed to industrial waste. Three potential isolates were identified as B. paramycoides, B. cereus, and B. tequilensis by 16S rRNA gene sequencing. At a concentration of 300µg/ml, these isolates demonstrated efficient FA degradation; 60-70% (at 300µg/ml FA) and 50-60%, (at 500µg/ml FA). Fourier-transform infrared (FTIR) spectroscopy and High-Performance Liquid Chromatography (HPLC) analysis further supported the result that the bacterial isolates consumed FA as the carbon source. Liquid chromatography-mass spectrometry (LC-MS) facilitates the detection of the major metabolic intermediate product in which FA gets transformed. The prominent peaks at 113 and 119m/z obtained in the MS spectra of the degraded FA samples indicated the possibility of the conversion of FA into furoic acid or levulinic acid. The phytotoxicity bioassay findings revealed the non-toxic nature of the bio-transformed products as compared to pure FA. This investigation presents the initial documentation of the FA degradative potential of Bacillus strains, thereby augmenting the understanding of the prospective implementation of Bacillus species in industrial waste treatment projects.

Entomopathogenic fungi of the genus Cordyceps biotransform zearalenone - metabolomic and proteomic backgrounds

Zearalenone (ZEN) is a mycotoxin that poses risks to animals and humans. Its metabolism in humans is well understood, but its biotransformation by fungi that co-exist in soil with Fusarium is not yet elucidated. In this study, the ability of the commonly used biocontrol agent Cordyceps to eliminate ZEN was evaluated. Overall, 19 phase I and II derivatives of ZEN biotransformation by reduction, oxidation, sulfonation, glycosylation and glucuronidation were detected. Importantly, new metabolites of ZEN biotransformation by fungi were discovered. C. fumosorosea and C. farinosa bioconverted ZEN to its oxidized and sulfonated form. Additionally, C. fumosorosea biotransformed zearalenol, the reduced metabolite of ZEN, via oxidation and sulfonation. Mechanistically, cytochrome P450 of the Cordyceps spp. catalyzed the oxidation of ZEN and its metabolites. Sulfate adenylyltransferase and adenylyl-sulfate kinase were responsible for sulfonation in the ZEN biotransformation pathway in C. fumosorosea. Additionally, enzymes that may be involved in the glycosylation and reduction were determined by proteomic research. Moreover, ZEN was found to decrease the level of beauvericin in C. fumosorosea, which could lead to the reduction in environmental pollution with this compound.

Impact of food preservatives based on immobilized phenolic compounds on an in vitro model of human gut microbiota

To address concerns about the biocompatibility of novel phenolic immobilization-based food preservatives, their impact on the composition and metabonomic profile of a defined community of human gut microbiota was evaluated. Three phenolics (eugenol, vanillin and ferulic acid) presented in two forms (free or immobilized on different supports) were tested at two concentration levels (0.5 and 2 mg/mL). Free eugenol was the phenolic with the greatest impact on gut microbiota, with a remarkable increase in the abundance of Lachnospiraceae and Akkermansiaceae families. In contrast, immobilized phenolics produced an increase in the abundance of Bacteroides with a reduction in the ratio of Firmicutes to Bacteroidetes. The metabonomic profile was also affected by free and immobilized phenolics differently in terms of fermentation by-products and phenolic biotransformation metabolites. Thus the results suggest the importance of evaluating the impact of new compounds or materials added to food on human gut microbiota and their potential use to modulate microbiota composition.

Peroxidases-based enticing biotechnological platforms for biodegradation and biotransformation of emerging contaminants

Rampant industrial boom, urbanization, and exponential population growth resulted in widespread environmental pollution, with water being one of the leading affected resources. All kinds of pollutants, including phenols, industrial dyes, antibiotics, pharmaceutically active residues, and persistent/volatile organic compounds, have a paramount effect, either directly or indirectly, on human health and aquatic entities. Strategies for affordable and efficient decontamination of these emerging pollutants have become the prime focus of academic researchers, industry, and government to constitute a sustainable human society. Classical treatment techniques for environmental contaminants are associated with several limitations, such as inefficiency, complex pretreatments, overall high process cost, high sludge generation, and highly toxic side-products formation. Enzymatic remediation is considered a green and ecologically friendlier method that holds considerable potential to mitigate any kinds of contaminating agents. Exploiting the potential of various peroxidases for pollution abatement is an emerging research area and has considerable advantages, such as efficiency and ease of handling, over other methods. This work is designed to provide recent progress in deploying peroxidases as green and versatile biocatalytic tools for the degradation and transformation of a spectrum of potentially hazardous environmental pollutants to broaden their scope for biotechnological and environmental purposes. More studies are required to explicate the degradation mechanisms, assess the toxicology levels of bio-transformed metabolites, and standardize the treatment strategies for economic viability.

Changes in the Plasma and Platelet Nitric Oxide Biotransformation Metabolites during Ischemic Stroke-A Dynamic Human LC/MS Metabolomic Study.

Despite improvement in the management of modifiable cardiovascular risk factors, ischemic stroke remains the leading cause of morbidity and mortality in the adult population. The aim of this study was to analyze the time-dependent dynamic differences in expression of the nitric oxide (NO) metabolic pathway in the platelet and plasma compartment between subjects with and without ischemic stroke. Additionally, the interplay between these parameters and platelet aggregation was investigated. A total of 418 patients in acute phase of non-cardioembolic stroke were investigated. Following the inclusion and exclusion criteria, finally 40 subjects with stroke and 39 demographically matched healthy participants were enrolled. Neurological physical examination, followed by assessment of the platelet and plasma levels of the nitric oxide synthase (NOS) inhibitors, including asymmetric dimethylarginine (ADMA) and symmetric dimethylarginine (SDMA), as well as NOS substrate-L-Arginine were performed dynamically three times within the first 24-h, then on the 3rd and 7th day after the stroke onset, which was compared with the healthy control. The platelet L-Arginine concentration was significantly higher on the 1st and 3rd day of stroke, while the plasma levels were significantly lower on exact days in comparison to the control. The competitive NOS-inhibitors in platelets were stably elevated in stroke subjects, whereas no significant differences in plasma compartment were noted. The arachidonic-acid-induced platelet aggregation was negatively associated with the platelet NOS substrate bioavailability, as assessed by the -ratio on the 3rd and 7th day. Subjects with non-cardioembolic ischemic stroke are characterized by elevated platelet levels of NOS inhibitors. Management of stroke results in increasing the platelet L-Arginine concentration and subsequent NO bioavailability in the platelet compartment.

Antioxidant potential of biotransformed green tea catechin metabolites and their impact on peripheral blood mononuclear cells

ObjectiveCatechin absorption, bioavailability, and distribution in human tissue and excretion were low. Colonic bacteria degrade catechin and few studies suggest that biotransformed metabolites may increase or decrease biological activities. However, the antioxidant potential of biotransformed metabolites is still unexplored. Hence, it is necessary to investigate the antioxidant potential of colonic bacterial transformed catechin metabolites. MethodsThe In vitro antioxidant potential of biotransformed catechin metabolites was assessed by DPPH, Hydrogen peroxide, and ABTS assay. Cytotoxic effect of biotransformed catechin metabolites on human mesenchymal stem cells and their response to antioxidant genes GPX1 and POR expression in human peripheral blood mononuclear cells (PBMC) were evaluated. ResultsFree radical scavenging activity of partially purified bacterial transformed catechin metabolites was low compared to pure catechin. The results revealed that variations in the antioxidant potential of pure catechin and colonic bacterial altered catechin metabolites. The IC50 value of partially purified biotransformed catechin metabolites exhibits free radical scavenging activity at high concentrations, while pure catechins act as a potent inhibitor at lower concentrations. The expression of GPX1 and POR gene increased four folds in a concentration dependent manner in pure catechin-treated cells. In contrast, partially purified biotransformed catechin metabolites treated cells expressed the downregulation of GPX1 and upregulation of POR gene. ConclusionTherefore, it can be concluded that the functional group alteration in biotransformed catechin metabolites may reduce the hydrogen donating ability, thus In vitro antioxidant potential of biotransformed catechin metabolites was low compared to pure catechin. Hence, GPX1 and POR exhibit different expression patterns in treated peripheral blood mononuclear cells. These findings suggest that pure catechin and colonic bacterial transformed catechin metabolites differed functionally.

Identification and Quantification of Both Methylation and Demethylation Biotransformation Metabolites of 5-Demethylsinensetin in Rats.

5-Demethylated polymethoxyflavones (5-OH PMFs) are the most unique monodemethylated PMFs with relatively low polarities and are proved to possess better anticancer and anti-inflammatory effects than their respective permethoxylated ones. However, their detailed in vivo metabolic fates have not been fully studied. 5-Demethylsinensetin (5-OH Sin), being one of the 5-demethylated citrus PMFs, was used in the present research to investigate its biotransformation in pharmacokinetics and excretion in rats. The results showed that 5-OH Sin was mostly accumulated in the large intestine, indicating its poor absorption in the small intestine. In addition, 5,3'-didemethylsinensetin and 5,4'-didemethylsinensetin were identified as two dominated metabolites of 5-OH Sin, and the C-3' position of 5-OH Sin was more facile to be demethylated in systemic circulation. Moreover, other than demethylation reactions, the methylation transformation of 5-OH Sin and its metabolites were also observed and quantified, suggesting that the bidirectional biotransformation between 5-OH Sin and its parent compound, Sin, occurred under in vivo conditions.

Neuroprotective Role of Microbial Biotransformed Metabolites of Sinapic Acid on Repetitive Traumatic Brain Injury in Rats

Sinapic acid biotransformation was studied and screened using eleven fungal cultures. Penicillium chermesinum AUMC 275, Aspergillus ochraceus AUMC 11328, and Paecilomyces variotii AUMC 5618 showed positive results in the production of metabolites. Sinapic acid biotransformation resulted in the production of gallic acid after 10 days incubation with P. chermesinum, protocatechuic acid after 7 days with A. ochraceus, and ferulic acid after 5 days with P. variotii. The investigated compounds' a

Ongoing and potential novel trends of pomegranate fruit peel; a comprehensive review of its health benefits and future perspectives as nutraceutical.

Pomegranate is an ancient shrub, globally distributed nowadays. It has been used in the middle east as a medicinal food and traditional medicine for thousands of years. Pomegranate peel (PP) constitutes about 50% of the total fruit, however, it has been previously regarded as a waste. Recent research points to PP as a rich source of phenolics (e.g., ellagitannins, flavonoids, and anthocyanins), polysaccharides, in addition to its biotransformed metabolites viz. urolithins making it a valuable waste with promising pharmacological actions. Compared to the pulp and the juice, PP exhibited stronger antioxidant and antimicrobial activities. Besides, it inhibited inflammation in several conditions, including colitis, arthritis, hepatitis, contact dermatitis, and lung inflammation. Moreover, it displayed anti-osteoporosis, anti-hyperglycemic, antidiabetic, antihypertensive, vasculoprotective, hepatoprotective, neuroprotective, and immunomodulatory effects. Additionally, it was effective as a prebiotic and in obesity control, besides it promoted wound healing. Furthermore, PP demonstrated anticancer effects against different cancer types, for example, colon, liver, thyroid, uterine, breast, bladder, prostate, leukemia, and osteosarcoma. Despite PP safety, it may interfere with the metabolism of other drugs because it inhibits cytochromes (CYP) changing their bioavailability, effectiveness, and toxicity. PP biowaste valorization not only avoids against its environmental and economic burden but can also provide a promising platform to produce novel or improved nutraceuticals. This study provides a comprehensive overview of PP biological activities with the reported action mechanisms related to its phytochemicals and further biotransformed metabolites inside the body. Future research prospects to unravel the merits of such waste and optimize its use are discussed. PRACTICAL APPLICATION: Pomegranate is widely distributed throughout the world. Although its peel was previously considered a waste, recent research regards it as a rich source of bioactive compounds with promising biological activities. Its recycling not only overcomes the bio-waste problems, but also provides a source of valuable compounds with several health benefits. In recent years, PP has been demonstrated to exhibit excellent pharmacological bioactivities, for example, antioxidant, anti-inflammatory, antimicrobial, antiosteoporosis, antihyperlipidemic, and anticancer activities. Its health-promoting power is mostly attributed to the phenolic and polysaccharide content, in addition to its amazing biotransformed metabolites. The underlying action mechanisms of such pharmacological activities are discussed and related to its chemical content. This review presents the latest research progress on the role of PP in the prevention and treatment of various chronic diseases, and its protective health effects for future research to be used in nutraceuticals.

Effect of Pyrethroid on Fertility of Adult Rats

Sixty  Adult female albino rats  were randomly divided into seven groups as fellow group (1) normal control, Inductive group (1) in this group animals received low dose  12 mg/kg) from permethrin,  Inductive group (2),in this group animals received high dose ( 75mg/kg.) from permethrin, group 4: treated group, animals received Vit E. (500 mg/ mg/Kg.) and then received low dose of permethrin (12 mg/k.) after half hour, group 5: treated group, animals received Vit E. (500 mg/ mg/Kg.) and then received high dose of permethrin (75 mg/k.) after half hour,Group 6: treated group, animals received Vit E. (250 mg/ mg/Kg) and then received low dose of permethrin (12 mg/k. ) after half hour and group 7: treated group, animals received Vit E. (250 mg/ mg/Kg.) and then received low dose of permethrin (75 mg/k. ) after half every doses received daily. At the end of the experimental period Blood samples were withdrawn,we determine triglyceride (TG), total cholesterol (TC), blood urea and creatinine. Also Estradiol and progesterone, total T3 , T4 and TSH levels  in plasma of female were measured. The excised ovary in each group used to measure MDA, as a marker for lipid peroxidation (LPO) and GSH content in ovary tissue . In the same time the ovarian tissue which were  removed after sacrifice,  it was used for histological examination for the evaluation and assessment of the d fibrous formation, in injuries tissue. permethrin with different doses (12 and 75 mg/kg  respectively) significantly (p≤ 0.05) increased the MDA  and significantly decreased (p< 0.05) in GSH content in ovary tissue versus control animal. Though treatment with vitamin E after permethrin with different dose (250 and 500 mg/kg respectively) significantly deceased the MDA content versus permethrin group (12 and 75 mg/kg  respectively) and Vit E treatment increased GSH content in a group treated with (permethrin 75 mg/kg b.w). The level of both serum estrogen (E2) and progesterone were significantly (p<0.05) decreased in a permethrin group in both doses (12 and 75 mg/kg ) when compared to control group. While, treatment with Vit E ameliorate the level of E2 and progesterone. Treating rats with permethrin led to a significant decrease in triiodothyronine (T3) and thyroxine (T4) as well as a significant increase of thyroid-stimulating hormone (TSH) when compared to control group. Administration of Vitamin E and permethrin caused an improvement in the levels ofT3 and T4 and TSH. This study was designed to elucidate premethrin effects at doses (12and75mg\kgw b) for 30 days, causing reproductive toxicity through several biotransformation metabolites processes. Focusing on ovarian injures and reproductive efficacy. treated ovarian tissue showed histological alterations represented by reduction number of different follicles, with absent mature follicles in high dose premethrin, or follicles without oocytes  and zone granulose residual with vacuolar change finally necrotic areas. Special stain for connective tissue fibersMasson's  trichrome, showed accumulation of fibrous proportional to the dose. while the Immuno- reactive of TNF-ᾳ assayment many necrotic area evidence of premethrin toxicity, also its dwindling effect on proliferating cell nuclear antigen (PCNA)which affected the efficiency of DNA, and consequently on the cell proliferative activity. In contrast, evaluate the effective therapeutic role of Vitamin E at doses (250 and 500 mg \kg bw),as an antioxidant clarified, which is explained by the  histological in the results.  This study reveals that chronic exposure to premerthin even   low dose alters ovarian cycle  activity . Hence, Premethrin exposure threat  fertility of humans and  animals.it is speculate that the positive effect of the Vitamin E by its antioxidant activity able to protect ovarian tissues the premerthin toxicicity

Chemical profiling of root bark extract from Oplopanax elatus and its in vitro biotransformation by human intestinal microbiota.

Oplopanax elatus (Nakai) Nakai, in the Araliaceae family, has been used in traditional Chinese medicine (TCM) to treat diseases as an adaptogen for thousands of years. This study established an ultra-performance liquid chromatography coupled with quadrupole time-of-flight tandem mass spectrometry (UPLC-Q-TOF/MS) method to identify chemical components and biotransformation metabolites of root bark extract from O. elatus. A total of 18 compounds were characterized in O. elatus extract, and 62 metabolites by human intestinal microbiota were detected. Two polyynes, falcarindiol and oplopandiol were recognized as the main components of O. elatus, whose metabolites are further illustrated. Several metabolic pathways were proposed to generate the detected metabolites, including methylation, hydrogenation, demethylation, dehydroxylation, and hydroxylation. These findings indicated that intestinal microbiota might play an essential role in mediating the bioactivity of O. elatus.

Biotransformation of Daidzein, Genistein, and Naringenin by Streptomyces Species Isolated from High-Altitude Soil of Nepal.

Flavonoids have achieved widespread importance in pharmaceutical, food, and cosmetics industries. Furthermore, modification of these naturally occurring flavonoids to structurally diverse compounds through whole cell biotransformation with enhanced biological activities has numerous biotechnological applications. The present study investigated the biotransformation potential of Streptomyces species isolated from a high-altitude-soil sample towards selected flavonoid molecules. The biotransformed metabolites were confirmed by comparing the HPLC chromatogram with authentic compounds and LC-MS/MS analysis. Of these isolates, Streptomyces species G-18 (Accession number: MW663767.1) catalyzed isoflavone molecules daidzein and genistein to produce hydroxylated products at 24 h of reaction condition in a whole cell system. The hydroxylation of daidzein (4′,7-dihydroxyisoflavone) was confirmed at 3′-position of the B ring to produce 3′,4′,7-trihydroxyisoflavone. In addition, Streptomyces species G-14 (Accession number: MW663770.1) and Streptomyces species S4L (Accession number: MW663769.1) also revealed the transformation of daidzein (4′,7-dihydroxyisoflavone) to hydroxy daidzein at a distinct position than that of G-18 isolates, whereas thee Streptomyces species S4L reaction mixture with naringenin as a substrate also revealed the hydroxylated product. Our results demonstrated that microorganisms isolated from different ecological niches have broad application.

Decolorization mechanism, identification of an FMN-dependent NADH-azoreductase from a moderately halotolerant Staphylococcus sp. MEH038S, and toxicity assessment of biotransformed metabolites.

The application of halotolerant microorganisms capable of decolorizing is attractive. Decolorization mechanism, the effect of different parameters on the decolorization percentage, and toxicity analysis of Reactive Black 5 before and after decolorization were investigated in the present study. The decolorization percentage for live cells of Staphylococcus sp. strain MEH038S was more than dead cells, which demonstrated that Reactive Black 5 was decolorized through the degradation process. The results confirmed that an FMN-dependent NADH-azoreductase gene was responsible for the decolorization and then was identified as Staphylococcus sp. EFS01 azoreductase from a moderately halotolerant Staphylococcus strain for the first time. The maximal decolorization of 98.15% was observed at pH 6.5 and 35 ° C for 50mg/L of Reactive Black 5. In addition, more than 90% decolorization was achieved with 5-40g/L of NaCl. The results of Gas chromatography-mass spectrometry and Fourier transform infrared spectroscopy showed that Reactive Black 5 was broken to the lower molecular weight compounds without any chromophoric azo groups. Phytotoxicity and fish toxicity proved that the biotransformed metabolites of Reactive Black 5degradation were more toxic than the original dye. The moderate halotolerant strain exhibited a remarkable decolorization capability and can be applied for textile wastewater treatment. PRACTITIONER POINTS: An azoreductase gene from a moderately halotolerant Staphylococcus was identified. More than 90% decolorization efficiency was observed under high-salt conditions. Biotransformed metabolites of RB5degradation were identified. Toxicity analysis of biotransformed metabolites was investigated.

Global-Scale Metabolomic Profiling of Human Hair for Simultaneous Monitoring of Endogenous Metabolome, Short- and Long-Term Exposome

Hair is a unique biological matrix that adsorbs short-term exposures (e. g., environmental contaminants and personal care products) on its surface and also embeds endogenous metabolites and long-term exposures in its matrix. In this work, we developed an untargeted metabolomics workflow to profile both temporal exposure chemicals and endogenous metabolites in the same hair sample. This analytical workflow begins with the extraction of short-term exposures from hair surfaces through washing. Further development of mechanical homogenization extracts endogenous metabolites and long-term exposures from the cleaned hair. Both solutions of hair wash and hair extract were analyzed using ultra-high-performance liquid chromatography-high-resolution mass spectrometry (UHPLC-HRMS)-based metabolomics for global-scale metabolic profiling. After analysis, raw data were processed using bioinformatic programs recently developed specifically for exposome research. Using optimized experimental conditions, we detected a total of 10,005 and 9,584 metabolic features from hair wash and extraction samples, respectively. Among them, 274 and 276 features can be definitively confirmed by MS2 spectral matching against spectral library, and an additional 3,356 and 3,079 features were tentatively confirmed as biotransformation metabolites. To demonstrate the performance of our hair metabolomics, we collected hair samples from three female volunteers and tested their hair metabolic changes before and after a 2-day exposure exercise. Our results show that 645 features from wash and 89 features from extract were significantly changed from the 2-day exposure. Altogether, this work provides a novel analytical approach to study the hair metabolome and exposome at a global scale, which can be implemented in a wide range of biological applications for a deeper understanding of the impact of environmental and genetic factors on human health.

Simultaneous decolorization/degradation of AB-113 and chromium(VI) removal by a salt-tolerant Klebsiella sp. AB-PR and detoxification of biotransformed-metabolites

Textile wastewater contains dyes, heavy metals, and salts. In this study, a salt-tolerant decolorizing strain was isolated from vermicompost and identified as Klebsiella sp. strain AB-PR by 16S ribosomal RNA gene analysis. Klebsiella sp. strain AB-PR was presented to simultaneously remove high concentrations of diazo dye and hexavalent chromium under saline conditions for the first time. Maximum decolorization of 95.50% was observed for 800 mg/L of Acid Blue-113 at pH 7 and temperature 35 °C with 5% inoculum size under static conditions. Also, decolorization efficiencies for 13 and 8 g/L of glucose and yeast extract as optimum carbon and nitrogen sources were 94.52 and 94.70%, respectively. Klebsiella sp. strain AB-PR could tolerate 50 g/L of NaCl and decolorize 94.18% of Acid Blue-113. The maximum decolorization and removal efficiencies for 800 mg/L of Acid Blue-113 and 30 mg/L of hexavalent chromium with 50 g/L of NaCl were 90.98 and 89.42%, respectively. Gas chromatography–mass spectrometry and Fourier transform-infrared spectroscopy studies showed that Acid Blue-113 was cleaved to the aromatic amines metabolites with lower molecular weight. The phyto-toxicity study exhibited that Acid Blue-113 was not toxic for Triticum aestivum and Maize. But after biological treatment, the toxicity was increased, and the seeds could not germinate. Therefore, ultraviolet-C/H2O2 was applied for the detoxification of biotransformed metabolites. After application of ultraviolet-C/H2O2, toxic aromatic metabolites were converted to nontoxic and linear organic products, and the seeds were germinated completely. Thus, the combined application of biological–chemical treatment is effective in the detoxification of textile wastewater.

Biotransformation of artemisinic acid to bioactive derivatives by endophytic Penicillium oxalicum B4 from Artemisia annua L.

As a biosynthetic precursor of the antimalarial drug artemisinin, artemisinic acid (AA) is abundant in Artemisia annua L. with a content of 8-10-fold higher than artemisinin, but less effective. In this study, the biotransformation of AA was carried out with an endophytic fungus Penicillium oxalicum B4 to extend its utility. After 10-day-culture of the endophyte with AA at 2 mg/mL, eight biotransformation metabolites were isolated from the culture broth, including five undescribed metabolites, namely 3α,14-dihydroxyartemisinic acid, 14-hydroxy-3-oxo-artemisinic acid, 15-hydroxy-3-oxo-artemisinic acid, 12, 15-artemisindioic acid and 1,2,3,6-tetradehydro-12, 15-artemisindioic acid. The fungal enzymes possess the selective capacity to hydroxylate, carbonylate and ketonize the allyl group of AA. The major biotransformation metabolite was the hydroxylated product 3-α-hydroxyartemisinic acid (33.3%) in the cultures of early stage (day 1–6), whereas most of the other biotransformation products were synthesized in the later stage (day 8–10). Compared with AA, some metabolites (3α,14-dihydroxyartemisinic acid, 15-hydroxy-3-oxo-artemisinic acid and 1,2,3,6-tetradehydro-12, 15-artemisindioic acid) possessed stronger cytotoxic activity to the human colon carcinoma cell line (LS174T) and promyelocytic leukemia cell line (HL-60). The metabolites 12, 15-artemisindioic acid and 3-α-hydroxyartemisinic acid exhibited significant inhibitory activity to the lipopolysaccharide-induced nitrite production of RAW 264.7 cells at 10.00 μM and 2.50 μM, respectively. The results demonstrated the potential of fungal endophytes on biotransforming AA to its bioactive derivatives.