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1.
PLoS One ; 19(6): e0304512, 2024.
Article En | MEDLINE | ID: mdl-38829838

The Organic Cation Transporter Novel 1 (OCTN1), also known as SLC22A4, is widely expressed in various human tissues, and involved in numerous physiological and pathological processes remains. It facilitates the transport of organic cations, zwitterions, with selectivity for positively charged solutes. Ergothioneine, an antioxidant compound, and acetylcholine (Ach) are among its substrates. Given the lack of experimentally solved structures of this protein, this study aimed at generating a reliable 3D model of OCTN1 to shed light on its substrate-binding preferences and the role of sodium in substrate recognition and transport. A chimeric model was built by grafting the large extracellular loop 1 (EL1) from an AlphaFold-generated model onto a homology model. Molecular dynamics simulations revealed domain-specific mobility, with EL1 exhibiting the highest impact on overall stability. Molecular docking simulations identified cytarabine and verapamil as highest affinity ligands, consistent with their known inhibitory effects on OCTN1. Furthermore, MM/GBSA analysis allowed the categorization of substrates into weak, good, and strong binders, with molecular weight strongly correlating with binding affinity to the recognition site. Key recognition residues, including Tyr211, Glu381, and Arg469, were identified through interaction analysis. Ach demonstrated a low interaction energy, supporting the hypothesis of its one-directional transport towards to outside of the membrane. Regarding the role of sodium, our model suggested the involvement of Glu381 in sodium binding. Molecular dynamics simulations of systems at increasing levels of Na+ concentrations revealed increased sodium occupancy around Glu381, supporting experimental data associating Na+ concentration to molecule transport. In conclusion, this study provides valuable insights into the 3D structure of OCTN1, its substrate-binding preferences, and the role of sodium in the recognition. These findings contribute to the understanding of OCTN1 involvement in various physiological and pathological processes and may have implications for drug development and disease management.


Molecular Docking Simulation , Molecular Dynamics Simulation , Organic Cation Transport Proteins , Humans , Organic Cation Transport Proteins/chemistry , Organic Cation Transport Proteins/metabolism , Organic Cation Transport Proteins/genetics , Symporters/chemistry , Symporters/metabolism , Binding Sites , Protein Binding , Ergothioneine/chemistry , Ergothioneine/metabolism , Sodium/metabolism , Sodium/chemistry , Computer Simulation , Acetylcholine/metabolism , Acetylcholine/chemistry , Ligands
2.
ACS Appl Mater Interfaces ; 16(23): 29917-29929, 2024 Jun 12.
Article En | MEDLINE | ID: mdl-38813785

Radiotherapy commonly causes damage to healthy tissues, particularly radiation-induced skin injury (RISI) that affects a significant majority of patients undergoing radiotherapy. Effective treatments for RISI are lacking. This study focuses on the pathogenesis of RISI, which primarily involves oxidative stress. Excessive reactive oxygen species (ROS) generation during radiation induces damage to biological macromolecules, triggering oxidative stress and inflammation. To address this, ergothioneine (EGT), a natural and biocompatibile thiol compound with excellent antioxidant activity, is explored as a potential radiation-protective agent. By utilizing its specific transport and absorption in the skin tissue, as well as its efficient and stable clearance of radiation-induced "ROS storm", EGT is combined with sodium hyaluronate (NaHA) to develop a novel radiation protective dressing suitable for the skin. This EGT-NaHA dressing demonstrates an effective ability to scavenge free radicals and reduce oxidative stress in vitro and in vivo, reducing cellular apoptosis and inflammation. These results demonstrate the protective properties of EGT against RISI, with far-reaching implications for research and development in the field of radioprotection.


Bandages , Ergothioneine , Hyaluronic Acid , Oxidative Stress , Radiation-Protective Agents , Skin , Hyaluronic Acid/chemistry , Hyaluronic Acid/pharmacology , Ergothioneine/pharmacology , Ergothioneine/chemistry , Animals , Oxidative Stress/drug effects , Oxidative Stress/radiation effects , Skin/drug effects , Skin/radiation effects , Skin/pathology , Mice , Humans , Radiation-Protective Agents/pharmacology , Radiation-Protective Agents/chemistry , Radiation-Protective Agents/therapeutic use , Reactive Oxygen Species/metabolism , Antioxidants/pharmacology , Antioxidants/chemistry , Apoptosis/drug effects , Apoptosis/radiation effects , Radiation Injuries/drug therapy , Radiation Injuries/prevention & control
3.
Chembiochem ; 25(9): e202400131, 2024 May 02.
Article En | MEDLINE | ID: mdl-38597743

Many actinobacterial species contain structural genes for iron-dependent enzymes that consume ergothioneine by way of O2-dependent dioxygenation. The resulting product ergothioneine sulfinic acid is stable under physiological conditions unless cleavage to sulfur dioxide and trimethyl histidine is catalyzed by a dedicated desulfinase. This report documents that two types of ergothioneine sulfinic desulfinases have evolved by convergent evolution. One type is related to metal-dependent decarboxylases while the other belongs to the superfamily of rhodanese-like enzymes. Pairs of ergothioneine dioxygenases (ETDO) and ergothioneine sulfinic acid desulfinase (ETSD) occur in thousands of sequenced actinobacteria, suggesting that oxidative ergothioneine degradation is a common activity in this phylum.


Ergothioneine , Ergothioneine/metabolism , Ergothioneine/chemistry , Actinobacteria/enzymology , Biocatalysis , Sulfinic Acids/chemistry , Sulfinic Acids/metabolism , Dioxygenases/metabolism , Dioxygenases/chemistry
4.
J Biol Chem ; 300(1): 105539, 2024 Jan.
Article En | MEDLINE | ID: mdl-38072054

L-ergothioneine is widely distributed among various microbes to regulate their physiology and pathogenicity within complex environments. One of the key steps in the ergothioneine-biosynthesis pathway, the C-S bond cleavage reaction, uses the pyridoxal 5'-phosphate dependent C-S lyase to produce the final product L-ergothioneine. Here, we present the crystallographic structure of the ergothioneine-biosynthesis C-S lyase EgtE from Mycobacterium smegmatis (MsEgtE) represents the first published structure of ergothioneine-biosynthesis C-S lyases in bacteria and shows the effects of active site residues on the enzymatic reaction. The MsEgtE and the previously reported ergothioneine-biosynthesis C-S lyase Egt2 from Neurospora crassa (NcEgt2) fold similarly. However, discrepancies arise in terms of substrate recognition, as observed through sequence and structure comparison of MsEgtE and NcEgt2. The structural-based sequence alignment of the ergothioneine-biosynthesis C-S lyase from fungi and bacteria shows clear distinctions among the recognized substrate residues, but Arg348 is critical and an extremely conserved residue for substrate recognition. The α14 helix is exclusively found in the bacteria EgtE, which represent the most significant difference between bacteria EgtE and fungi Egt2, possibly resulting from the convergent evolution of bacteria and fungi.


Ergothioneine , Lyases , Mycobacterium , Ergothioneine/chemistry , Ergothioneine/metabolism , Fungi/metabolism , Lyases/chemistry , Lyases/metabolism , Mycobacterium/metabolism , Mycobacterium smegmatis/chemistry , Mycobacterium smegmatis/enzymology , Models, Molecular , Protein Structure, Quaternary , Protein Structure, Tertiary
5.
Int J Biol Macromol ; 256(Pt 2): 128428, 2024 Jan.
Article En | MEDLINE | ID: mdl-38013086

Selenoneine (SEN) is a natural histidine derivative with radical-scavenging activity and shows higher antioxidant potential than its sulfur-containing isolog ergothioneine (EGT). Recently, the SEN biosynthetic pathway in Variovorax paradoxus was reported. Resembling EGT biosynthesis, the committed step of SEN synthesis is catalyzed by a nonheme Fe-dependent oxygenase termed SenA. This enzyme catalyzes oxidative carbon­selenium (C-Se) bond formation to conjugate N-α-trimethyl histidine (TMH) and selenosugar to yield selenoxide; the process parallels the EGT biosynthetic route, in which sulfoxide synthases known as EgtB members catalyze the conjugation of TMH and cysteine or γ-glutamylcysteine to afford sulfoxides. Here, we report the crystal structures of SenA and its complex with TMH and thioglucose (SGlc), an analog of selenoglucose (SeGlc) at high resolution. The overall structure of SenA adopts the archetypical fold of EgtB, which comprises a DinB-like domain and an FGE-like domain. While the TMH-binding site is highly conserved to that of EgtB, a various substrate-enzyme interaction network in the selenosugar-binding site of SenA features a number of water-mediated hydrogen bonds. The obtained structural information is beneficial for understanding the mechanism of SenA-mediated C-Se bond formation.


Ergothioneine , Organoselenium Compounds , Histidine , Iron , Oxygenases , Ergothioneine/chemistry , Ergothioneine/metabolism
6.
Brain Res ; 1824: 148693, 2024 02 01.
Article En | MEDLINE | ID: mdl-38036238

Oxidative stress can upset the antioxidant balance and cause accelerated aging including neurodegenerative diseases and decline in physiological function. Therefore, an antioxidant-rich diet plays a crucial role in healthy aging. This study aimed to identify and quantify mushrooms with the highest ergothioneine content through HPLC analysis and evaluate their anti-aging potential as a natural antioxidant and antisenescence in HT22 cells. Among the 14 evaluated mushroom species, Lentinula edodes (LE), shiitake mushroom contains the highest ergothioneine content and hence was used for the in-vitro studies. The cells were preincubated with ethanolic extract of ergothioneine-rich mushroom and the equimolar concentration of EGT on t-BHP-induced senescence HT22 cells. The extract was analyzed for its free radical scavenging properties using DPPH and ABTS methods. Then, the neuroprotective effect was conducted by measuring the cell viability using MTT. Senescence-associated markers and ROS staining were also analyzed. Our results revealed that a low dose of t-BHP reduces cell viability and induces senescence in HT22 cells as determined through ß-galactosidase staining and expressions of P16INK4a, P21CIPL which are the markers of cellular senescence. However, the pretreatment with ethanolic extract of LE for 8 h significantly improved the cell viability, reversed the t-BHP-induced cellular senescence in the neuronal cells, and reduced the reactive oxygen species visualized through DCFH-DA staining. These results suggest that ergothioneine-rich mushroom is a potential candidate for anti-aging exploration through the elimination of senescent cells.


Agaricales , Ergothioneine , Ergothioneine/pharmacology , Ergothioneine/chemistry , Antioxidants/pharmacology , Antioxidants/metabolism , Agaricales/chemistry , Agaricales/metabolism , Cellular Senescence
7.
Free Radic Biol Med ; 198: 12-26, 2023 03.
Article En | MEDLINE | ID: mdl-36736443

Recently we have uncovered a non-enzymatic multi-step cycle for the regeneration of ergothioneine (ET), after reaction with noxious singlet oxygen (1O2), by glutathione (GSH). When living cells were loaded with ET labeled with deuterium and N-15 atoms (D5-ET) and exposed to light in the presence of a photosensitizer, no loss of deuterium at position 5 of the imidazole ring was observed, in contradiction to our previous mechanistic proposal. Therefore, it was necessary to reexamine the in vitro products of ET and 1O2 by liquid chromatography coupled to high resolution mass spectrometry. Pure 1O2 was generated by thermolysis at 37 °C of the endoperoxide DHPNO2. The use of D5-ET enabled us to revise and extend the reaction scheme. On the main pathway, 1O2 attacks the imidazole ring, and the hydroperoxide intermediates are reduced rapidly by ET or GSH via different mechanisms. The intramolecular water elimination from the 5-hydroperoxide described previously is slower and not a part of the cycle. On another side path, 1O2 attacks the sulfur of ET to form a sulfine (S-oxide). The reduction of the sulfine also allows for the complete regeneration of ET. Experiments with methanol instead of water as solvent revealed that, in the absence of GSH, ET was attacked 6 times more frequently at the ring than at the sulfur. In the presence of 1 mM GSH or higher, both side paths were abandoned. ET efficiently captures 1O2 with its ring and can then be regenerated to a large extent by GSH, without enzyme involvement.


Ergothioneine , Ergothioneine/chemistry , Singlet Oxygen/chemistry , Hydrogen Peroxide/chemistry , Deuterium , Glutathione/metabolism , Imidazoles , Water , Oxygen
8.
J Agric Food Chem ; 71(1): 671-679, 2023 Jan 11.
Article En | MEDLINE | ID: mdl-36571834

Ergothioneine (ERG) is an unusual sulfur-containing amino acid with antioxidant activity that can be synthesized by certain bacteria and fungi. Microbial fermentation is a promising method for ERG production. In this study, the bifunctional enzyme methyltransferase-sulfoxide synthase NcEgt1 from Neurospora crassa was truncated to obtain sulfoxide synthase TNcEgt1, which showed a higher expression level in Escherichia coli BL21(DE3). Then, the genes egtD encoding methyltransferase EgtD and egtE encoding C-S lyase EgtE from Mycobacterium smegmatis were cloned with TncEgt1 into E. coli BL21(DE3) to produce 70 mg/L ERG. To improve ERG production, TNcEgt1 and EgtD were modified, and the resulting mutants were screened with an established high-throughput method which could directly analyze the ERG content in culture broths. After several rounds of mutation and screening, the optimal mutant MD4 was obtained and produced 290 mg/L ERG. Furthermore, a fed-batch culture was conducted in a 5 L bioreactor. After optimizing the fermentation process, the ERG yield reached 5.4 g/L after 94 h of cultivation supplemented with amino acids and glycerol, which is the highest ERG yield reported to date. The results showed that ERG production was significantly improved by modifying the key enzymes, and the engineered strains constructed in this study have potential industrial application prospects.


Ergothioneine , Antioxidants/metabolism , Bacteria/genetics , Ergothioneine/chemistry , Escherichia coli/genetics , Escherichia coli/metabolism , Fermentation , Metabolic Engineering , Methyltransferases/metabolism
9.
Sci Rep ; 11(1): 22240, 2021 11 15.
Article En | MEDLINE | ID: mdl-34782676

Ergothioneine (EGT) is a low molecular weight histidine betaine essential in all domains of life but only synthesized by selected few organisms. Synthesis of EGT by Mycobacterium tuberculosis (M. tb) is critical for maintaining bioenergetic homeostasis and protecting the bacterium from alkylating agents, oxidative stress, and anti-tubercular drugs. EgtD, an S-adenosylmethionine-dependent methyltransferase (AdoMet), catalyzes the trimethylation of L-Histidine to initiate EGT biosynthesis and this reaction has been shown to be essential for EGT production in mycobacteria and for long-term infection of murine macrophages by M. tb. In this work, library screening and structure-guided strategies identified multiple classes of M. tb EgtD inhibitors that bind in various regions of the enzyme active site. X-ray crystal structures of EgtD-inhibitor complexes confirm that L-Histidine analogs bind solely to the L-Histidine binding site while drug-like inhibitors, such as TGX-221, and S-Glycyl-H-1152 span both the L-Histidine and AdoMet binding sites. These enzyme-inhibitor complexes provide detailed structural information of compound scaffolds useful for developing more potent inhibitors that could shorten Tuberculosis treatment regimens by weakening important bacterial defenses.


Antitubercular Agents/chemistry , Betaine/analogs & derivatives , Binding Sites , Biosynthetic Pathways/drug effects , Ergothioneine/chemistry , Histidine/analogs & derivatives , Models, Molecular , Mycobacterium tuberculosis/drug effects , Antitubercular Agents/pharmacology , Betaine/chemistry , Betaine/metabolism , Dose-Response Relationship, Drug , Ergothioneine/biosynthesis , Histidine/chemistry , Histidine/metabolism , Histidine/pharmacology , Molecular Conformation , Molecular Structure , Mycobacterium tuberculosis/metabolism , Structure-Activity Relationship
10.
Sci Rep ; 11(1): 18450, 2021 09 16.
Article En | MEDLINE | ID: mdl-34531467

Ergothioneine (ERGO) is a rare amino acid mostly found in fungi, including mushrooms, with recognized antioxidant activity to protect tissues from damage by reactive oxygen species (ROS) components. Prior to this publication, the biodistribution of ERGO has been performed solely in vitro using extracted tissues. The aim of this study was to develop a feasible chemistry for the synthesis of an ERGO PET radioligand, [11C]ERGO, to facilitate in vivo study. The radioligand probe was synthesized with identical structure to ERGO by employing an orthogonal protection/deprotection approach. [11C]methylation of the precursor was performed via [11C]CH3OTf to provide [11C]ERGO radioligand. The [11C]ERGO was isolated by RP-HPLC with a molar activity of 690 TBq/mmol. To demonstrate the biodistribution of the radioligand, we administered approximately 37 MBq/0.1 mL in 5XFAD mice, a mouse model of Alzheimer's disease via the tail vein. The distribution of ERGO in the brain was monitored using 90-min dynamic PET scans. The delivery and specific retention of [11C]ERGO in an LPS-mediated neuroinflammation mouse model was also demonstrated. For the pharmacokinetic study, the concentration of the compound in the serum started to decrease 10 min after injection while starting to distribute in other peripheral tissues. In particular, a significant amount of the compound was found in the eyes and small intestine. The radioligand was also distributed in several regions of the brain of 5XFAD mice, and the signal remained strong 30 min post-injection. This is the first time the biodistribution of this antioxidant and rare amino acid has been demonstrated in a preclinical mouse model in a highly sensitive and non-invasive manner.


Antioxidants/pharmacokinetics , Ergothioneine/pharmacokinetics , Positron-Emission Tomography/methods , Radiopharmaceuticals/pharmacokinetics , Animals , Antioxidants/chemistry , Carbon Radioisotopes/chemistry , Ergothioneine/chemistry , Mice , Mice, Inbred C57BL , Radiopharmaceuticals/chemistry , Tissue Distribution
11.
Int J Mol Sci ; 22(6)2021 Mar 23.
Article En | MEDLINE | ID: mdl-33806754

Medicinal use of mushrooms has been documented since ancient times, and in the modern world, mushrooms have a longstanding history of use in Eastern medicine. Recent interest in plant-based diets in Westernized countries has brought increasing attention to the use of mushrooms and mushroom-derived compounds in the prevention and treatment of chronic diseases. Edible mushrooms are the most abundant food sources of the modified amino acid, ergothioneine. This compound has been shown to accumulate in almost all cells and tissues, but preferentially in those exposed to oxidative stress and injury. The demonstrated cytoprotectant effect of ergothioneine has led many to suggest a potential therapeutic role for this compound in chronic conditions that involve ongoing oxidative stress and inflammation, including cardiovascular and metabolic diseases. However, the in vivo effects of ergothioneine and its underlying therapeutic mechanisms in the whole organism are not as clear. Moreover, there are no well-defined, clinical prevention and intervention trials of ergothioneine in chronic disease. This review highlights the cellular and molecular mechanisms of action of ergothioneine and its potential as a Traditional, Complementary and Alternative Medicine for the promotion of cardiometabolic health and the management of the most common manifestations of cardiometabolic disease.


Agaricales/chemistry , Biological Products/pharmacology , Energy Metabolism/drug effects , Ergothioneine/pharmacology , Heart/drug effects , Myocardium/metabolism , Animals , Anti-Inflammatory Agents/chemistry , Anti-Inflammatory Agents/pharmacology , Antioxidants/chemistry , Antioxidants/pharmacology , Biological Products/chemistry , Dietary Supplements , Ergothioneine/chemistry , Humans
12.
Biosci Biotechnol Biochem ; 85(5): 1175-1182, 2021 Apr 24.
Article En | MEDLINE | ID: mdl-33686392

Ergothioneine (ERGO), a thiohistidine betaine, exists in various fungi, plants, and animals. Humans take in ERGO from their diet. ERGO is a strong biological antioxidant, but there are only a limited number of reports about its redox mechanism. The purpose of this study was to clarify the oxidation mechanism of ERGO. Reactions of ERGO with chemical oxidants were performed. The oxidation products of ERGO were analyzed by nuclear magnetic resonance and liquid chromatography-mass spectrometry (LC-MS). The major product of oxidation of ERGO by hydrogen peroxide in physiological conditions was identified as hercynine (histidine betaine). One molecule of ERGO was able to reduce 2 molecules of hydrogen peroxide. Hercynine was found to react with the more potent oxidant hypochlorite. One unstable decomposition product was detected by LC-MS. As a result, a mechanism of oxidation of ERGO, and hence its physiological antioxidant activity, was developed.


Antioxidants/chemistry , Betaine/analogs & derivatives , Calcium Compounds/chemistry , Ergothioneine/chemistry , Histidine/analogs & derivatives , Hydrogen Peroxide/chemistry , Oxidants/chemistry , Betaine/chemistry , Chromatography, Liquid , Histidine/chemistry , Kinetics , Oxidation-Reduction , Solutions , Tandem Mass Spectrometry , Water/chemistry
13.
Curr Mol Pharmacol ; 14(2): 220-233, 2021.
Article En | MEDLINE | ID: mdl-32048982

BACKGROUND: The enhancement of learning and memory through food-derived ingredients is of great interest to healthy individuals as well as those with diseases. Ergothioneine (ERGO) is a hydrophilic antioxidant highly contained in edible golden oyster mushrooms (Pleurotus cornucopiae var. citrinopileatus), and systemically absorbed by its specific transporter, carnitine/organic cation transporter OCTN1/SLC22A4. OBJECTIVE: This study aims to examine the possible enhancement of object recognition memory by oral administration of ERGO in normal mice. METHODS: Novel object recognition test, spatial recognition test, LC-MS/MS, Golgi staining, neuronal culture, western blotting, immunocytochemistry, and quantitative RT-PCR were utilized. RESULT: After oral administration of ERGO (at a dose of 1-50 mg/kg) three times per week for two weeks in ICR mice, the novel object recognition test revealed a longer exploration time for the novel object than for the familiar object. After oral administration of ERGO, the spatial recognition test also revealed a longer exploration time for the spatially moved object than the unmoved one in mice fed ERGO-free diet. The discrimination index was significantly higher in the ERGO-treated group than the control in both behavioral tests. ERGO administration led to an increase in its concentration in the plasma and hippocampus. The systemic concentration reached was relevant to those found in humans after oral ERGO administration. Golgi staining revealed that ERGO administration increased the number of matured spines in the hippocampus. Exposure of cultured hippocampal neurons to ERGO elevated the expression of the synapse formation marker, synapsin I. This elevation of synapsin I was inhibited by the tropomyosin receptor kinase inhibitor, K252a. Treatment with ERGO also increased the expression of neurotrophin-3 and -5, and phosphorylated mammalian target of rapamycin in hippocampal neurons. CONCLUSION: Oral intake of ERGO may enhance object recognition memory at its plasma concentration achievable in humans, and this enhancement effect could occur, at least in part, through the promotion of neuronal maturation in the hippocampus.


Antioxidants/chemistry , Behavior, Animal/drug effects , Enzyme Inhibitors/chemistry , Ergothioneine/chemistry , Nutritional Physiological Phenomena/drug effects , Pleurotus/chemistry , Administration, Oral , Animals , Antioxidants/administration & dosage , Antioxidants/analysis , Carbazoles/pharmacology , Chromatography, High Pressure Liquid , Dose-Response Relationship, Drug , Drug Discovery , Enzyme Inhibitors/administration & dosage , Enzyme Inhibitors/blood , Ergothioneine/administration & dosage , Ergothioneine/blood , Hippocampus/metabolism , Humans , Indole Alkaloids/pharmacology , Male , Mice, Inbred ICR , Neurogenesis/drug effects , Neurons/metabolism , Synapsins/metabolism , Tandem Mass Spectrometry
14.
Microb Cell Fact ; 19(1): 164, 2020 Aug 18.
Article En | MEDLINE | ID: mdl-32811496

BACKGROUND: Ergothioneine (EGT) has a unique antioxidant ability and diverse beneficial effects on human health. But the content of EGT is very low in its natural producing organisms such as Mycobacterium smegmatis and mushrooms. Therefore, it is necessary to highly efficient heterologous production of EGT in food-grade yeasts such as Saccharomyces cerevisiae. RESULTS: Two EGT biosynthetic genes were cloned from the mushroom Grifola frondosa and successfully heterologously expressed in Saccharomyces cerevisiae EC1118 strain in this study. By optimization of the fermentation conditions of the engineered strain S. cerevisiae EC1118, the 11.80 mg/L of EGT production was obtained. With daily addition of 1% glycerol to the culture medium in the fermentation process, the EGT production of the engineered strain S. cerevisiae EC1118 can reach up to 20.61 mg/L. CONCLUSION: A successful EGT de novo biosynthetic system of S. cerevisiae containing only two genes from mushroom Grifola frondosa was developed in this study. This system provides promising prospects for the large scales production of EGT for human health.


Agaricales/genetics , Ergothioneine/biosynthesis , Glycerol/metabolism , Saccharomyces cerevisiae/genetics , Saccharomyces cerevisiae/metabolism , Antioxidants/chemistry , Ergothioneine/chemistry , Fermentation , Gene Expression Regulation, Bacterial , Genes, Fungal , Industrial Microbiology , Microorganisms, Genetically-Modified
15.
J Nutr Biochem ; 84: 108453, 2020 10.
Article En | MEDLINE | ID: mdl-32653808

There is evidence from both in vitro and animal models that the consumption of edible mushrooms has beneficial effects on health. It is unclear whether similar effects exist in humans and which bioactive compounds are present. This review synthesises the evidence on the world's most commonly consumed mushroom, Agaricus bisporus to (i) examine its effect on human health outcomes; and (ii) determine the nutrient density of its bioactive compounds, which may explain their health effects. A systematic literature search was conducted on the consumption of A. bisporus, without date and study design limits. Bioactive compounds included ergosterol, ergothioneine, flavonoids, glucans and chitin. Two authors independently identified studies for inclusion and assessed methodological quality. Beneficial effects of A. bisporus on metabolic syndrome, immune function, gastrointestinal health and cancer, with the strongest evidence for the improvement in Vitamin D status in humans, were found. Ultraviolet B (UVB) exposed mushrooms may increase and maintain serum 25(OH)D levels to a similar degree as vitamin D supplements. A. bisporus contain beta-glucans, ergosterol, ergothioneine, vitamin D and an antioxidant compound usually reported as flavonoids; with varying concentrations depending on the type of mushroom, cooking method and duration, and UVB exposure. Further research is required to fully elucidate the bioactive compounds in mushrooms using vigorous analytical methods and expand the immunological markers being tested. To enable findings to be adopted into clinical practice and public health initiatives, replication of existing studies in different population groups is required to confirm the impact of A. bisporus on human health.


Agaricus , Biological Products/pharmacology , Agaricus/chemistry , Animals , Biological Products/chemistry , Biological Products/therapeutic use , Ergosterol/chemistry , Ergosterol/pharmacology , Ergosterol/therapeutic use , Ergothioneine/chemistry , Ergothioneine/pharmacology , Ergothioneine/therapeutic use , Flavonoids/chemistry , Flavonoids/pharmacology , Flavonoids/therapeutic use , Glucans/chemistry , Glucans/pharmacology , Glucans/therapeutic use , Humans
16.
Chembiochem ; 21(20): 2908-2911, 2020 10 15.
Article En | MEDLINE | ID: mdl-32614492

Ergothioneine has emerged as a crucial cytoprotectant in the pathogenic lifestyle of Mycobacterium tuberculosis. Production of this antioxidant from primary metabolites may be regulated by phosphorylation of Thr213 in the active site of the methyltransferase EgtD. The structure of mycobacterial EgtD suggests that this post-translational modification would require a large-scale change in conformation to make the active-site residue accessible to a protein kinase. In this report, we show that, under in vitro conditions, EgtD is not a substrate of protein kinase PknD.


Ergothioneine/biosynthesis , Methyltransferases/metabolism , Mycobacterium tuberculosis/enzymology , Ergothioneine/chemistry , Methyltransferases/chemistry , Models, Molecular , Molecular Conformation , Protein Kinases/metabolism , Substrate Specificity
17.
Int J Med Mushrooms ; 22(2): 171-181, 2020.
Article En | MEDLINE | ID: mdl-32479005

Ergothioneine is a natural 2-thiol-amidazole amino acid that plays an important role in inflammation, depression, and cardiovascular disease. Flammulina velutipes is a common basidiomycete mushroom rich in ergothioneine (EGT). However, the biosynthetic pathway of EGT in F. velutipes is still unclear. In this study, the F. velutipes ergothioneine biosynthetic gene 1 (Fvegtl), F. velutipes ergothioneine biosynthetic gene 2 (Fvegt2), and F. velutipes ergothioneine biosynthetic gene 3 (Fvegt3) were cloned and expressed, and the activities of the proteins encoded by these three genes (FvEgt1, F. velutipes ergothioneine biosynthase 1; FvEgt2, F. velutipes ergothioneine biosynthase 2; and FvEgt3, F. velutipes ergothioneine biosynthase 3) were identified. The results showed that FvEgtl not only has the function of methyltransferase, but also has the function of hercynlcysteineteine sulfoxide (Hersul) synthase, which can catalyze the production of Hersul from histidine and cysteine in F. velutipes. FvEgt2 and FvEgt3 are two functionally different cysteine desulfurase enzymes. Among them, FvEgt2 is a cysteine-cysteine desulfurase-which catalyzes the activation of the S-H bond on cysteine, while FvEgt3 is a pyridoxal phosphate (PLP)-dependent cysteine desulfurase responsible for catalyzing the production of ketimine complex. Our results show that FvEgt1/FvEgt2/FvEgt3 can simultaneously catalyze the production of EGT by histidine, cysteine, and pyridoxal phosphate. Collectively, the in vitro synthesis of EGT in the edible fungus F. velutipes was first achieved, which laid the foundation for the biological production of EGT.


Antioxidants/metabolism , Biosynthetic Pathways/genetics , Ergothioneine/metabolism , Flammulina/chemistry , Agaricales , Antioxidants/chemistry , Cysteine/metabolism , Ergothioneine/chemistry , Escherichia coli/genetics , Escherichia coli/metabolism , Flammulina/enzymology , Flammulina/genetics , Gene Expression , Histidine/metabolism , Pyridoxal Phosphate/metabolism
18.
Int J Med Mushrooms ; 22(3): 211-220, 2020.
Article En | MEDLINE | ID: mdl-32479016

The naturally occurring amino acid ergothioneine (EGT) has excellent free radical scavenging ability, which was not different to ascorbic acid. The IC50 values for EGT scavenging hydrogen peroxide, hydroxyl radicals, and superoxide anions were 11.65 ± 0.31, 70.31 ± 1.59, and 160.44 ± 0.32 µg/mL, respectively. The EGT concentration in different species of mushrooms was significantly different (p < 0.05), but it was not significantly related to the ability of the mushrooms to scavenge reactive oxygen species (p > 0.05). After isolating EGT from mushrooms, we demonstrated that the antioxidant ability of EGT accounts for about 25% of the total antioxidant ability of the extract. We studied the stability of EGT and found that it has excellent light, thermal, and acid-base stability. However, the presence of Cu2+ decreased the concentration of EGT. Unlike EGT, the thermal stability of the EGT extracted from Pleurotus citrinopileatus (PEGT) was not as good as EGT, while long-term high-temperature heating caused a decrease in the concentration of PEGT. The results of our study provide a basis for further investigating EGT from mushrooms for research and development.


Agaricales/chemistry , Antioxidants/metabolism , Ergothioneine/chemistry , Food Analysis , Hot Temperature , Inhibitory Concentration 50 , Reactive Oxygen Species/metabolism , Superoxides
19.
Chemistry ; 26(6): 1328-1334, 2020 Jan 27.
Article En | MEDLINE | ID: mdl-31545545

Sulfoxide synthases are non-heme iron enzymes that participate in the biosynthesis of thiohistidines, such as ergothioneine and ovothiol A. The sulfoxide synthase EgtB from Chloracidobacterium thermophilum (CthEgtB) catalyzes oxidative coupling between the side chains of N-α-trimethyl histidine (TMH) and cysteine (Cys) in a reaction that entails complete reduction of molecular oxygen, carbon-sulfur (C-S) and sulfur-oxygen (S-O) bond formation as well as carbon-hydrogen (C-H) bond cleavage. In this report, we show that CthEgtB and other bacterial sulfoxide synthases cannot efficiently accept selenocysteine (SeCys) as a substrate in place of cysteine. In contrast, the sulfoxide synthase from the filamentous fungus Chaetomium thermophilum (CthEgt1) catalyzes C-S and C-Se bond formation at almost equal efficiency. We discuss evidence suggesting that this functional difference between bacterial and fungal sulfoxide synthases emerges from different modes of oxygen activation.


Acidobacteria/enzymology , Bacterial Proteins/antagonists & inhibitors , Fungal Proteins/antagonists & inhibitors , Selenocysteine/chemistry , Bacterial Proteins/metabolism , Binding Sites , Binding, Competitive , Biocatalysis , Catalytic Domain , Cysteine Dioxygenase/antagonists & inhibitors , Cysteine Dioxygenase/metabolism , Ergothioneine/chemistry , Ergothioneine/metabolism , Fungal Proteins/metabolism , Kinetics , Molecular Dynamics Simulation , Mycobacteriaceae/enzymology , Selenocysteine/metabolism
20.
Chemistry ; 25(44): 10298-10303, 2019 Aug 06.
Article En | MEDLINE | ID: mdl-31188501

Ergothioneine is a sulfur-containing histidine derivative that emerges from microbial biosynthesis and enters the human body through intestinal uptake and regulated distribution into specific tissues. Although the proteins involved in biosynthesis and uptake are well characterized, less is known about the degradative pathways of ergothioneine. This report describes the crystal structure of the active form of ergothionase from the oral pathogen Treponema denticola complexed with the substrate analogue desmethyl-ergothioneine sulfonic acid. This enzyme catalyzes the 1,2-elimination of trimethylamine from ergothioneine and ergothioneine sulfonic acid by using a unique mode of substrate activation combined with acid/base catalysis. This structural and mechanistic investigation revealed four essential catalytic residues, which are strictly conserved in homologous proteins from common gastrointestinal bacteria and numerous pathogenic bacteria, suggesting that bacterial activity may play an important role in determining the availability of ergothioneine in healthy and diseased human tissue.


Bacterial Proteins/chemistry , Carbon-Nitrogen Lyases/chemistry , Ergothioneine/chemistry , Treponema denticola/enzymology , Catalysis , Catalytic Domain , Crystallization , Models, Molecular , Protein Conformation
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