Recent progress in ergothioneine biosynthesis: a review / 生物工程学报

Ergothioneine is a multifunctional physiological cytoprotector, with broad application in foods, beverage, medicine, cosmetics and so on. Biosynthesis is an increasingly favored method in the production of ergothioneine. This paper summarizes the new progress in the identification of key pathways, the mining of key enzymes, and the development of natural edible mushroom species and high-yield engineering strains for ergothioneine biosynthesis in recent years. Through this review, we aim to reveal the molecular mechanism of ergothioneine biosynthesis and then employ the methods of fermentation engineering, metabolic engineering, and synthetic biology to greatly increase the yield of ergothioneine.

Construction and optimization of ergothioneine-producing Escherichia coli / 生物工程学报

Ergothioneine (ERG) is a natural antioxidant that has been widely used in the fields of food, medicine and cosmetics. Compared with traditional plant extraction and chemical synthesis approaches, microbial synthesis of ergothioneine has many advantages, such as the short production cycle and low cost, and thus has attracted intensive attention. In order to engineer an ergothioneine high-yielding Escherichia coli strain, the ergothioneine synthesis gene cluster egtABCDE from Mycobacterium smegmatis and egt1 from Schizosaccharomyces pombe were introduced into E. coli BL21(DE3) to generate a strain E1-A1 harboring the ergothioneine biosynthesis pathway. As a result, (95.58±3.2) mg/L ergothioneine was produced in flask cultures. To further increase ergothioneine yield, the relevant enzymes for biosynthesis of histidine, methionine, and cysteine, the three precursor amino acids of ergothioneine, were overexpressed. Individual overexpression of serAT410STOP and thrA resulted in an ergothioneine titer of (134.83±4.22) mg/L and (130.26±3.34) mg/L, respectively, while co-overexpression of serAT410STOP and thrA increased the production of ergothioneine to (144.97±5.40) mg/L. Eventually, by adopting a fed-batch fermentation strategy in 3 L fermenter, the optimized strain E1-A1-thrA-serA* produced 548.75 mg/L and 710.53 mg/L ergothioneine in glucose inorganic salt medium and rich medium, respectively.

Ergothioneine Contents in Fruiting Bodies and Their Enhancement in Mycelial Cultures by the Addition of Methionine

The levels of ergothioneine (ERG), which have been shown to act as an excellent antioxidant, were determined in both fruiting bodies and mycelia of various mushroom species. We found that ERG accumulated at different levels in fruiting bodies of mushrooms and showed up to a 92.3-fold difference between mushrooms. We also found that ERG accumulated at higher levels in mycelia than in fruiting bodies of economically important mushroom species such as Ganoderma neo-japonicum, G. applanatum and Paecilomyces tenuipes. The addition of 2 mM methionine (Met) to mycelial culture medium increased the ERG contents in most mushroom species tested, indicating that Met is a good additive to enhance the ERG levels in a variety of mushroom species. Taking these results into consideration, we suggest that the addition of Met to the mycelial culture medium is an efficient way to enhance the antioxidant properties in economically important mushroom species.

Effect of l-carnitine and deferoxamine on hepatotoxicity, erythrocyte ergothioneine and testicular testosterone synthesis in sodiumvalproate-treated rats

The aim of the present study was to evaluate the effect of sodium valproate [VPA] as anantiepileptic drug on liver and erythrocyte oxidative state and on testicular testosterone synthesis. The therapeuticefficacy of either L-carnitine or deferoxamine [DFO] was assessed in such situations. This study included 108 albino rats divided into control groups, single dose VPA treated groups withand without L-carnitine or DFO treatment and repeated dose VPA treated groups with and without L-carnitine orDFO treatment. Liver triglycerides and malondialdehyde levels and catalase and glutathione S transferaseactivities as well as erythrocyte ergothioneine were estimated. In testes, P450cl7 and 17/3 hydroxysteroiddehydrogenase activities were measured as testosterone synthesis. Single and repeated VPA administration caused an increase in liver triglycerides, malondialdehyde,catalase and glutathione S transferase and a decrease in erythrocyte ergothioneine. Only repeated VPAadministration could decrease testicular testosterone synthesis. Co-administration of L-carnitine or DFO withVPA resulted in improvement of all the studied parameters. These data support the oxidative stress as a possible mechanism implicated in VPA-induced liver anderythrocyte damage. Moreover, VPA could alter testosterone synthesis giving a possible link between malereproductive function and VPA. The protective influence of L-carnitine and DFO may be partly mediated by theirantioxidant effects and by improving gonadal testosterone synthesi

L-ergothioneine level in red blood cells of healthy human males in the Western province of Saudi Arabia

Ergothioneine is widely distributed in biological systems, particularly in red blood cells of animals. However, it's functional role in human body is not well understood. In order to investigate the biochemical effect of L-ergothioneine, its concentration changes in human blood with respect to ages in healthy individuals was first investigated. L-ergothioneine concentrations in the blood of Saudi males from western province at different stages of life were measured by the procedure of Carlsson et al., 1974. At early stages of life (1-10 years), the concentrations of LER is 1.5-2.0 mg/100 ml. It increases gradually at the age of 11-18 years where it reaches the maximum value of 3.7 mg/100 ml. Then, it declines gradually to 3.0-2.3 mg/ 100 ml during the period of 19-50 years. An increase in the level of LER (2.8 mg/100 ml) was seen at the age of 51+.