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Barry Halliwell - One of the best experts on this subject based on the ideXlab platform.
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could Ergothioneine aid in the treatment of coronavirus patients
Antioxidants, 2020Co-Authors: Irwin K Cheah, Barry HalliwellAbstract:Infection with SARS-CoV-2 causes the coronavirus infectious disease 2019 (COVID-19), a pandemic that has, at present, infected more than 11 million people globally. Some COVID-19 patients develop a severe and critical illness, spurred on by excessive inflammation that can lead to respiratory or multiorgan failure. Numerous studies have established the unique array of cytoprotective properties of the dietary amino acid Ergothioneine. Based on studies in a range of in vitro and in vivo models, Ergothioneine has exhibited the ability to modulate inflammation, scavenge free radicals, protect against acute respiratory distress syndrome, prevent endothelial dysfunction, protect against ischemia and reperfusion injury, protect against neuronal damage, counteract iron dysregulation, hinder lung and liver fibrosis, and mitigate damage to the lungs, kidneys, liver, gastrointestinal tract, and testis, amongst many others. When compiled, this evidence suggests that Ergothioneine has a potential application in the treatment of the underlying pathology of COVID-19. We propose that Ergothioneine could be used as a therapeutic to reduce the severity and mortality of COVID-19, especially in the elderly and those with underlying health conditions. This review presents evidence to support that proposal.
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specificity of the Ergothioneine transporter natively expressed in hela cells
Biochemical and Biophysical Research Communications, 2019Co-Authors: Robert Alan Jappy Tucker, Irwin K Cheah, Barry HalliwellAbstract:Abstract Ergothioneine is a biologically important compound that has been shown to be transported by the organic cation transporter novel type 1 (OCTN1). Following this discovery, a variety of alternate functions for OCTN1 have been suggested including an integral function in the extra-neuronal cholinergic system. The present study reaffirms the primacy of Ergothioneine over these alternate substrates using natively expressed OCTN1 in HeLa cells. Besides the general transport inhibitors, quinidine, verapamil and pyrilamine no other putative substrate inhibited Ergothioneine transport significantly, with only a slight inhibition demonstrated by carnitine. Even compounds structurally similar to Ergothioneine failed to inhibit Ergothioneine uptake, suggesting high selectivity of OCTN1. Ergothioneine was found to be avidly accumulated even at low concentrations (300 nM) by HeLa cells.
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Ergothioneine a diet derived antioxidant with therapeutic potential
FEBS Letters, 2018Co-Authors: Barry Halliwell, Irwin K Cheah, Richard Ming Yi TangAbstract:Ergothioneine is a thiol/thione molecule synthesised only by some fungi and bacteria. Nonetheless, it is avidly taken up from the diet by humans and other animals through a transporter, OCTN1, and accumulates to high levels in certain tissues. Ergothioneine is not rapidly metabolised, or excreted in urine and is present in many, if not all, human tissues and body fluids. Ergothioneine has powerful antioxidant and cytoprotective properties in vitro and there is evidence that the body may concentrate it at sites of tissue injury by raising OCTN1 levels. Decreased blood and/or plasma levels of Ergothioneine have been observed in some diseases, suggesting that a deficiency could be relevant to the disease onset or progression. This brief Review explores the possible roles of Ergothioneine in human health and disease.
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Ergothioneine – a diet‐derived antioxidant with therapeutic potential
FEBS Letters, 2018Co-Authors: Barry Halliwell, Irwin K Cheah, Richard Ming Yi TangAbstract:Ergothioneine is a thiol/thione molecule synthesised only by some fungi and bacteria. Nonetheless, it is avidly taken up from the diet by humans and other animals through a transporter, OCTN1, and accumulates to high levels in certain tissues. Ergothioneine is not rapidly metabolised, or excreted in urine and is present in many, if not all, human tissues and body fluids. Ergothioneine has powerful antioxidant and cytoprotective properties in vitro and there is evidence that the body may concentrate it at sites of tissue injury by raising OCTN1 levels. Decreased blood and/or plasma levels of Ergothioneine have been observed in some diseases, suggesting that a deficiency could be relevant to the disease onset or progression. This brief Review explores the possible roles of Ergothioneine in human health and disease.
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antioxidant action of Ergothioneine assessment of its ability to scavenge peroxynitrite
Biochemical and Biophysical Research Communications, 1997Co-Authors: Okezie I Aruoma, Matthew Whiteman, Timothy G England, Barry HalliwellAbstract:Abstract The superoxide radical (O•−2) and nitric oxide (NO•) combine very rapidly to form peroxynitrite (ONOO−), a reactive tissue damaging nitrogen species thought to be involved in the pathology of several chronic diseases. The natural product Ergothioneine protects against the nitration of tyrosine and the inactivation of α1-antiproteinase by ONOO−. Ergothioneine merits further investigation as a biological and therapeutic antioxidant agent.
Okezie I Aruoma - One of the best experts on this subject based on the ideXlab platform.
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l Ergothioneine modulates oxidative damage in the kidney and liver of rats in vivo studies upon the profile of polyunsaturated fatty acids
Clinical Nutrition, 2004Co-Authors: Monica Deiana, Antonella Rosa, Viviana Casu, R Piga, Assunta M Dessi, Okezie I AruomaAbstract:BACKGROUND & AIMS: L-Ergothioneine is a fungal metabolite exhibiting antioxidant functions in cells. The aim of this study was to assess the effect of oral administration of L-Ergothioneine on the oxidative damage in vivo caused by the Fenton reagent ferric-nitrilotriacetate. METHODS: Rats were supplemented with L-ergo prior to the administration of acute dose of ferric-nitrilotriacetate. Kidney and liver levels of L-Ergothioneine, glutathione, alpha-tocopherol, polyunsaturated fatty acids and conjugated dienes were assessed. RESULTS: Oral administration of 70 mg L-ergo/kg body weight of rats for 7 days prior to the injection of ferric-nitrilotriacetate protected the fatty acids against oxidation, with notable protections directed to: 20:5 (eicosapentaenoic acid) (23%), 22:6 (docosahexaenoinic acid) (30%), 20:3 n6 (eicosatrienoic acid) (22%), 20:4 (arachidonic acid) (25%), 18:2 linoleic acid (25%) and 18:1 oleic acid (14%) in the kidney. The protection of 20:5, 20:3 n6 and 18:1 in the liver by 32%, 20% and 11%, respectively, were statistically significant. L-Ergothioneine significantly reduced kidney and liver levels of conjugated dienes and conserved the concentrations of alpha-tocopherol and glutathione in the kidney and liver in the ferric-nitrilotriacetate/L-Ergothioneine treated rats. CONCLUSION: Supplementation with L-Ergothioneine not only protects the organs against the lipid peroxidation but conserves the consumption of endogenous glutathione and alpha-tocopherol. However consumption of mushrooms may have better promise as dietary sources of L-Ergothioneine to humans.
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Ergothioneine inhibits oxidative stress and tnf α induced nf κb activation and interleukin 8 release in alveolar epithelial cells
Biochemical and Biophysical Research Communications, 2003Co-Authors: Irfan Rahman, Peter S Gilmour, Saibal K Biswas, Luis A Jimenez, Frank Antonicelli, Okezie I AruomaAbstract:Abstract Oxidants and inflammatory mediators such as tumour necrosis factor-α (TNF-α) activate transcription factors such as NF-κB. Interleukin-8 (IL-8) is a ubiquitous inflammatory chemokine that mediates a multitude of inflammatory events in the lung. Ergothioneine is a naturally occurring thiol compound, which possesses antioxidant property. The aim of this study was to determine whether Ergothioneine can inhibit the hydrogen peroxide (H2O2)- and TNF-α-mediated activation of NF-κB and the release of IL-8 in human alveolar epithelial cells (A549). Treatment of A549 cells with H2O2 (100 μM) and TNF-α (10 ng/ml) significantly increased NF-κB activation using a reporter assay. Ergothioneine inhibited both H2O2- and TNF-α-mediated activation of NF-κB. Both H2O2 and TNF-α significantly increased IL-8 release, which was inhibited by pre-treatment of A549 cells with Ergothioneine compared to the control untreated cells. Ergothioneine also abolished the transcriptional activation of IL-8 in an IL-8-chloramphenicol acetyltransferase (CAT) reporter system, transfected into A549 cells. This indicates a molecular mechanism for the anti-inflammatory effects of Ergothioneine.
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Ergothioneine inhibits oxidative stress- and TNF-alpha-induced NF-kappa B activation and interleukin-8 release in alveolar epithelial cells.
Biochemical and biophysical research communications, 2003Co-Authors: Irfan Rahman, Peter S Gilmour, Saibal K Biswas, Luis A Jimenez, Frank Antonicelli, Okezie I AruomaAbstract:Oxidants and inflammatory mediators such as tumour necrosis factor-alpha (TNF-alpha) activate transcription factors such as NF-kappa B. Interleukin-8 (IL-8) is a ubiquitous inflammatory chemokine that mediates a multitude of inflammatory events in the lung. Ergothioneine is a naturally occurring thiol compound, which possesses antioxidant property. The aim of this study was to determine whether Ergothioneine can inhibit the hydrogen peroxide (H(2)O(2))- and TNF-alpha-mediated activation of NF-kappa B and the release of IL-8 in human alveolar epithelial cells (A549). Treatment of A549 cells with H(2)O(2) (100 microM) and TNF-alpha (10 ng/ml) significantly increased NF-kappa B activation using a reporter assay. Ergothioneine inhibited both H(2)O(2)- and TNF-alpha-mediated activation of NF-kappa B. Both H(2)O(2) and TNF-alpha significantly increased IL-8 release, which was inhibited by pre-treatment of A549 cells with Ergothioneine compared to the control untreated cells. Ergothioneine also abolished the transcriptional activation of IL-8 in an IL-8-chloramphenicol acetyltransferase (CAT) reporter system, transfected into A549 cells. This indicates a molecular mechanism for the anti-inflammatory effects of Ergothioneine.
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effects of Ergothioneine on diabetic embryopathy in pregnant rats
Food and Chemical Toxicology, 2002Co-Authors: Maria V Guijarro, Okezie I Aruoma, A Indart, M Viana, B BonetAbstract:Abstract The natural antioxidant Ergothioneine (2-mercaptohistidine trimethylbetaine) is a fungal metabolite and found in most plant and animal tissues. The effect of Ergothioneine on diabetic embryopathy in rats was assessed. Supplementation of diabetic pregnant rats with l -Ergothioneine (1.147 mg/kg body weight) daily for the first 11.5 days of pregnancy reduced the rate of embryo malformations, to values similar to the non-diabetic animals. The Ergothioneine had no effect on the plasma glucose levels, both in diabetic and control animals. We conclude that the inhibition of the glucose-mediated free radical dependent embryo malformation by Ergothioneine is an important antioxidant prophylactic mechanism, which when combined with vitamin E could benefit the management of diabetic embryopathy.
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antioxidant action of Ergothioneine assessment of its ability to scavenge peroxynitrite
Biochemical and Biophysical Research Communications, 1997Co-Authors: Okezie I Aruoma, Matthew Whiteman, Timothy G England, Barry HalliwellAbstract:Abstract The superoxide radical (O•−2) and nitric oxide (NO•) combine very rapidly to form peroxynitrite (ONOO−), a reactive tissue damaging nitrogen species thought to be involved in the pathology of several chronic diseases. The natural product Ergothioneine protects against the nitration of tyrosine and the inactivation of α1-antiproteinase by ONOO−. Ergothioneine merits further investigation as a biological and therapeutic antioxidant agent.
Noriyoshi Isozumi - One of the best experts on this subject based on the ideXlab platform.
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impairment of the carnitine organic cation transporter 1 Ergothioneine axis is mediated by intestinal transporter dysfunction in chronic kidney disease
Kidney International, 2017Co-Authors: Yasuyuki Shinozaki, Kengo Furuichi, Tadashi Toyama, Shinji Kitajima, Akinori Hara, Yasunori Iwata, Norihiko Sakai, Miho Shimizu, Shuichi Kaneko, Noriyoshi IsozumiAbstract:Carnitine/organic cation transporter 1 (OCTN1) is a specific transporter of the food-derived antioxidant Ergothioneine. Ergothioneine is absorbed by intestinal OCTN1, distributed through the bloodstream, and incorporated into each organ by OCTN1. OCTN1 expression is upregulated in injured tissues, and promotes Ergothioneine uptake to reduce further damage caused by oxidative stress. However, the role of the OCTN1–Ergothioneine axis in kidney–intestine cross-talk and chronic kidney disease (CKD) progression remains unclear. Here we assessed Ergothioneine uptake via intestinal OCTN1 and confirmed the expression of OCTN1. The ability of OCTN1 to absorb Ergothioneine was diminished in mice with CKD. In combination with OCTN1 dysfunction, OCTN1 localization on the intestinal apical cellular membrane was disturbed in mice with CKD. Proteomic analysis, RT-PCR, Western blotting, and immunohistochemistry revealed that PDZ (PSD95, Dlg, and ZO1), a PDZK1 domain-containing protein that regulates the localization of transporters, was decreased in mice with CKD. Decreased intestinal Ergothioneine uptake from food decreased Ergothioneine levels in the blood of mice with CKD. Despite increased OCTN1 expression and Ergothioneine uptake into the kidneys of mice with CKD, Ergothioneine levels did not increase. To identify the role of the OCTN1-Ergothioneine axis in CKD, we evaluated kidney damage and oxidative stress in OCTN1-knockout mice with CKD and found that kidney fibrosis worsened. Oxidative stress indicators were increased in OCTN1-knockout mice. Moreover, Ergothioneine levels in the blood of patients with CKD decreased, which were restored after kidney transplantation. Thus, a novel inter-organ interaction mediated by transporters is associated with CKD progression.
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Impairment of the carnitine/organic cation transporter 1-Ergothioneine axis is mediated by intestinal transporter dysfunction in chronic kidney disease.
Kidney International, 2017Co-Authors: Yasuyuki Shinozaki, Kengo Furuichi, Tadashi Toyama, Shinji Kitajima, Akinori Hara, Yasunori Iwata, Norihiko Sakai, Miho Shimizu, Shuichi Kaneko, Noriyoshi IsozumiAbstract:Carnitine/organic cation transporter 1 (OCTN1) is a specific transporter of the food-derived antioxidant Ergothioneine. Ergothioneine is absorbed by intestinal OCTN1, distributed through the bloodstream, and incorporated into each organ by OCTN1. OCTN1 expression is upregulated in injured tissues, and promotes Ergothioneine uptake to reduce further damage caused by oxidative stress. However, the role of the OCTN1–Ergothioneine axis in kidney–intestine cross-talk and chronic kidney disease (CKD) progression remains unclear. Here we assessed Ergothioneine uptake via intestinal OCTN1 and confirmed the expression of OCTN1. The ability of OCTN1 to absorb Ergothioneine was diminished in mice with CKD. In combination with OCTN1 dysfunction, OCTN1 localization on the intestinal apical cellular membrane was disturbed in mice with CKD. Proteomic analysis, RT-PCR, Western blotting, and immunohistochemistry revealed that PDZ (PSD95, Dlg, and ZO1), a PDZK1 domain-containing protein that regulates the localization of transporters, was decreased in mice with CKD. Decreased intestinal Ergothioneine uptake from food decreased Ergothioneine levels in the blood of mice with CKD. Despite increased OCTN1 expression and Ergothioneine uptake into the kidneys of mice with CKD, Ergothioneine levels did not increase. To identify the role of the OCTN1-Ergothioneine axis in CKD, we evaluated kidney damage and oxidative stress in OCTN1-knockout mice with CKD and found that kidney fibrosis worsened. Oxidative stress indicators were increased in OCTN1-knockout mice. Moreover, Ergothioneine levels in the blood of patients with CKD decreased, which were restored after kidney transplantation. Thus, a novel inter-organ interaction mediated by transporters is associated with CKD progression.
Florian P. Seebeck - One of the best experts on this subject based on the ideXlab platform.
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Convergent Evolution of Fungal Cysteine Dioxygenases
ChemBioChem, 2020Co-Authors: Sebastian Flückiger, Nico V Igareta, Florian P. SeebeckAbstract:Cupin-type cysteine dioxygenases (CDO) are non-heme iron enzymes that occur in animals, plants, bacteria and in filamentous fungi. In this report show that agaricomycetes contain an entirely unrelated type of CDOs that emerged by convergent evolution from enzymes involved in the biosynthesis of Ergothioneine. The activity of this CDO type is dependent on the Ergothioneine precursor N-α-trimethylhistidine. The metabolic link between Ergothioneine production and cysteine oxidation suggest that the two processes may be part of the same chemical response in fungi - for example against oxidative stress.
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Structural basis of Ergothioneine biosynthesis.
Current Opinion in Structural Biology, 2020Co-Authors: Anja R. Stampfli, Wulf Blankenfeldt, Florian P. SeebeckAbstract:: Ergothioneine is a sulfur-containing histidine derivative synthesized by many bacteria and most fungi but it also finds its way into human tissue by way of specific absorption from the diet. The precise role of Ergothioneine is not yet known but there is growing evidence that it plays a role as an antioxidant protecting human cells from oxidative stress and pathogenic bacteria from host defenses. In this review we highlight recent advances in understanding the structural basis of Ergothioneine biosynthesis. In addition to unusual carbon-sulfur bond forming enzymology this research has revealed that Ergothioneine biosynthesis has emerged at least three times by independent molecular evolution.
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Structure and Mechanism of Ergothionase from Treponema denticola
Chemistry: A European Journal, 2019Co-Authors: Alice Maurer, Florian Leisinger, Florian P. SeebeckAbstract:: 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.
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structural and mechanistic basis for anaerobic Ergothioneine biosynthesis
Journal of the American Chemical Society, 2019Co-Authors: Florian Leisinger, Reto Burn, Marcel Meury, Peer Lukat, Florian P. SeebeckAbstract:Ergothioneine is an emergent factor in cellular redox biochemistry in humans and pathogenic bacteria. Broad consensus has formed around the idea that Ergothioneine protects cells against reactive oxygen species. The recent discovery that anaerobic microorganisms make the same metabolite using oxygen-independent chemistry indicates that Ergothioneine also plays physiological roles under anoxic conditions. In this report, we describe the crystal structure of the anaerobic Ergothioneine biosynthetic enzyme EanB from green sulfur bacterium Chlorobium limicola. This enzyme catalyzes the oxidative sulfurization of N-α-trimethyl histidine. On the basis of structural and kinetic evidence, we describe the catalytic mechanism of this unusual C–S bond-forming reaction. Significant active-site conservation among distant EanB homologues suggests that the oxidative sulfurization of heterocyclic substrates may occur in a broad range of bacteria.
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Inhibition and Regulation of the Ergothioneine Biosynthetic Methyltransferase EgtD.
ACS Chemical Biology, 2018Co-Authors: Laetitia Misson, Wulf Blankenfeldt, Reto Burn, Julia Hildesheim, Mariia A. Beliaeva, Florian P. SeebeckAbstract:Ergothioneine is an emerging factor in cellular redox homeostasis in bacteria, fungi, plants, and animals. Reports that Ergothioneine biosynthesis may be important for the pathogenicity of bacteria and fungi raise the question as to how this pathway is regulated and whether the corresponding enzymes may be therapeutic targets. The first step in Ergothioneine biosynthesis is catalyzed by the methyltransferase EgtD that converts histidine into N-α-trimethylhistidine. This report examines the kinetic, thermodynamic and structural basis for substrate, product, and inhibitor binding by EgtD from Mycobacterium smegmatis. This study reveals an unprecedented substrate binding mechanism and a fine-tuned affinity landscape as determinants for product specificity and product inhibition. Both properties are evolved features that optimize the function of EgtD in the context of cellular Ergothioneine production. On the basis of these findings, we developed a series of simple histidine derivatives that inhibit methyltra...
Douglas B. Kell - One of the best experts on this subject based on the ideXlab platform.
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effect of l Ergothioneine on the metabolic plasma profile of the rupp rat model of pre eclampsia
PLOS ONE, 2020Co-Authors: Audeclaire Morillon, Douglas B. Kell, Rachel D Williamson, Philip N Baker, Louise C Kenny, Jane A English, Fergus P Mccarthy, Cathal MccarthyAbstract:Introduction Pre-eclampsia is a major cause of maternal and fetal mortality and morbidity worldwide. Its pathophysiology remains unclear, but mitochondrial dysfunction and oxidative stress have been implicated. L-Ergothioneine is a naturally occurring, water-soluble betaine, that has demonstrated antioxidant properties. Using the reduced uterine perfusion pressure (RUPP) rat model of pre-eclampsia, this study aimed to define the plasma metabolic profile following treatment with L-Ergothioneine. Methods The effect of L-Ergothioneine (ET) treatment was explored using in vivo treatment in rats: Sham control (SC, n = 5), RUPP control (RC, n = 5), Sham +ET (ST, n = 5), RUPP +ET (RT, n = 5). Differential expression of plasma metabolites were obtained using untargeted liquid chromatography coupled to mass spectrometry. Statistical analysis was performed on normalised data comparing RC to SC, RT to RC, and RT to ST. Metabolites significantly altered (FDR < 0.05) were identified through database search. Results We report significantly lower levels of L-palmitoylcarnitine in RC compared to SC, a fatty acyl substrate involved in beta-oxidation in the mitochondria. We report that a metabolite that has been associated with oxidative stress (Glutamylcysteine) was detected at significantly higher levels in RT vs RC and RT vs ST. Five metabolites associated with inflammation were significantly lower in RT vs RC and three metabolites in RT vs ST, demonstrating the anti-inflammatory effects of ET in the RUPP rat model of pre-eclampsia. Conclusions L-Ergothioneine may help preserve mitochondrial function by increasing antioxidant levels, and reducing inflammatory responses associated with pre-eclampsia. This study shows the potential of L-Ergothioneine as a treatment for pre-eclampsia.
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engineering the yeast saccharomyces cerevisiae for the production of l Ergothioneine
Frontiers in Bioengineering and Biotechnology, 2019Co-Authors: Steven Axel Van Der Hoek, Behrooz Darbani, Karolina E. Zugaj, Bala K. Prabhala, Mathias Bernfried Biron, Milica Randelovic, Jacqueline B. Medina, Douglas B. Kell, Irina BorodinaAbstract:: L-(+)-Ergothioneine (ERG) is an unusual, naturally occurring antioxidant nutraceutical that has been shown to help reduce cellular oxidative damage. Humans do not biosynthesise ERG, but acquire it from their diet; it exploits a specific transporter (SLC22A4) for its uptake. ERG is considered to be a nutraceutical and possible vitamin that is involved in the maintenance of health, and seems to be at too low a concentration in several diseases in vivo. Ergothioneine is thus a potentially useful dietary supplement. Present methods of commercial production rely on extraction from natural sources or on chemical synthesis. Here we describe the engineering of the baker's yeast Saccharomyces cerevisiae to produce Ergothioneine by fermentation in defined media. After integrating combinations of ERG biosynthetic pathways from different organisms, we screened yeast strains for their production of ERG. The highest-producing strain was also engineered with known Ergothioneine transporters. The effect of amino acid supplementation of the medium was investigated and the nitrogen metabolism of S. cerevisiae was altered by knock-out of TOR1 or YIH1. We also optimized the media composition using fractional factorial methods. Our optimal strategy led to a titer of 598 ± 18 mg/L Ergothioneine in fed-batch culture in 1 L bioreactors. Because S. cerevisiae is a GRAS ("generally recognized as safe") organism that is widely used for nutraceutical production, this work provides a promising process for the biosynthetic production of ERG.
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Engineering the yeast Saccharomyces cerevisiae for the production of L-(+)-Ergothioneine
bioRxiv, 2019Co-Authors: Steven Axel Van Der Hoek, Behrooz Darbani, Karolina E. Zugaj, Bala K. Prabhala, Mathias Bernfried Biron, Milica Randelovic, Jacqueline B. Medina, Douglas B. Kell, Irina BorodinaAbstract:L-(+)-Ergothioneine is an unusual, naturally occurring antioxidant nutraceutical that has been shown to help reduce cellular oxidative damage. Humans do not biosynthesise it, so can acquire it only from their diet; it exploits a specific transporter (SLC22A4) for its uptake. ERG is considered to be a nutraceutical and possible vitamin that is involved in the maintenance of health, and seems to be at too low a concentration in several diseases in vivo. Ergothioneine is thus a potentially useful dietary supplement. Present methods of commercial production rely on extraction from natural sources or on chemical synthesis. Here we describe the engineering of the baker9s yeast Saccharomyces cerevisiae to produce Ergothioneine by fermentation in defined media. After integrating combinations of ERG biosynthetic pathways from different organisms, we screened yeast strains for their production of ERG. The highest-producing strain was also engineered with known Ergothioneine transporters. The effect of amino acid supplementation of the medium was investigated and the nitrogen metabolism of S. cerevisiae was altered by knock-out of TOR1 or YIH1. We also optimized the media composition using fractional factorial methods. Our optimal strategy led to a titer of 598 ± 18 mg/L Ergothioneine in fed-batch culture in bioreactors. Because S. cerevisiae is a GRAS (9generally recognised as safe9) organism that is widely used for nutraceutical production, this work provides a promising process for the biosynthetic production of ERG.