Medium optimization based on yeast's elemental composition for glutathione production in Saccharomyces cerevisiae

Schmacht, Maximilian; Lorenz, Eric; Ståhl, Ulf; Senz, Martin

doi:10.1016/j.jbiosc.2016.12.011

Cited by 14 publications

(10 citation statements)

References 45 publications

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“…The addition of the precursor amino acids, such as cysteine, glycine and glutamate was investigated by many researchers to increase the intracellular GSH content (Alfafara et al, 1992 a and b;Wang et al, 2012;Schmacht et al, 2017), and it was concluded that cysteine represents the key amino acid for GSH production and is the most common inducer for GSH overproduction (Schmacht et al, 2017). It is important to nd out the optimal concentration of cysteine because this amino acid is the substrate of gamma-glutamyl cysteine synthetase; which is the rst enzyme committed in GSH biosynthesis, however, at the same time, cysteine can act as growth inhibitor.…”

Section: The Effect Of Addition Time Of Cysteinementioning

confidence: 99%

Bioprocess optimization of glutathione production by Saccharomyces boulardii: biochemical characterization of glutathione peroxidase

et al. 2021

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The well-known probiotic GRAS Saccharomyces boulardii (CNCM I-745) was used for the rst time to produce glutathione (GSH). The culture conditions affecting GSH biosynthesis were screened using a Plackett-Burman design (PBD). Analyzing the regression coe cients for 12 tested variables; 6 of them, including yeast extract, glucose, peptone and cysteine; temperature and agitation rate had a positive signi cant effect on GSH production with a maximum production of 192 mg/L. The impact of addition time of cysteine was investigated in 19 experiments during the growth time course (0-36 h), the best addition time was 8h post-inoculation producing 235 mg/L of GSH. The most signi cant variables were further explored at 5-levels using Central Composite Rotatable Design (CCRD), giving a maximum production of GSH (552 mg/L). Using ba ed asks, the GSH was increased to 730 mg/L, i.e 1.32-folds increment than obtained using CCRD. The two rate-limiting genes of GSH biosynthesis "γ-glutamyl cysteine synthetase (gsh1) and GSH-synthetase (gsh2) were ampli ed and sequenced to validate the GSH biosynthetic potency of S. boulardii. The sequences of genes showed 99% similarity with gsh1 and gsh2 genes of S. cerevisiae. Glutathione peroxidase was puri ed and characterized from S. boulardii with molecular mass and subunit structure of 80 kDa and 35 kDa as revealed from native and SDS-PAGE, ensuring its homodimeric identity. The activity of GPx was reduced by 2.5-folds upon demetallization con rming its metalloproteinic identity. The enzyme was strongly inhibited by hydroxylamine and DTNB, ensuring the implication of surface lysine and cysteine residues on the enzyme active site domains.

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Section: The Effect Of Addition Time Of Cysteinementioning

confidence: 99%

Bioprocess optimization of glutathione production by Saccharomyces boulardii: biochemical characterization of glutathione peroxidase

et al. 2021

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“…Применяются различные пути накопления глутатиона в дрожжах: использование мутагенных штаммов с высокой накопительной способностью к глутатиону, внесение аминокислот-прекурсоров [23,34].…”

Section: результаты и их обсуждениеunclassified

“…Рассматривалось влияние источников азота на синтез глутатиона дрожжами. Сульфат аммония, являясь источником как азота, так и серы, необходим для увеличения плотности биомассы и образования глутатиона [23].…”

Section: результаты и их обсуждениеunclassified

“…Добавление цистеина в экспоненциальную фазу замедляет рост клеток за счет появления эффекта Кребтри из-за того, что образование этанола ведет к подавлению цикла трикарбоновых кислот. Внесение L-цистеина в стационарную фазу положительно сказывается на содержании глутатиона (1,76 % на СВ) [23]. Имеет значение также и дозировка внесения L-цистеина [24].…”

Section: результаты и их обсуждениеunclassified

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Yeasts as a Glutathione Producer

Meledina

Morozov

Davydenko³

et al. 2020

Food Processing: Techniques and Technology

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Introduction. Yeast is a fast-growing single-celled microorganism and an inexpensive source of various biologically active substances, such as antioxidants, e.g. Glutathione (GSH). Antioxidant properties are determined by the presence of sulfhydryl group. The global demand for glutathione is estimated to exceed 9 billion USD at the expense not only of pure crystalized glutathione, but also of glutathione-enriched yeast extracts. In the food industry, glutathione is used to improve the quality of the dough and enhance the taste of various products. The present research featured domestic and foreign studies on the content of glutathione in yeast, methods of biosynthesis, and antioxidant properties. Results and discussion. The content of glutathione ranges from 0.1 to 1% per completely dry biomass (CDB) in wild yeast strains. The fermentative method for the accumulation of glutathione is based on the optimization of the nutrient medium and the use of glutathione precursors, i.e. cysteine, glutamic acid, and glycine. Thus, this method makes it possible to double the content of intracellular glutathione in certain cultivation conditions. The use of non-directed mutagenesis methods can increase glutathione synthesis up to 5% in separate mutant strains, although the mechanism of synthesis is not always clear under such conditions. However, up to 2.27% of glutathione is being formed under directed change of the genome. In addition, the level of glutathione in cells increases under the influence of certain physical factors. For example, glutathione biosynthesis increases by 39% if yeast is exposed to a magnetic field. The enzymatic method requires maintaining the following factors: the presence of precursors (L-glutamic acid, L-cysteine, glycine), ATP, Mg2+ ions to activate GSH1 and GSH2, the pH of the medium, and the introduction of the necessary enzymes into the bioreactor. Hiwever, this method is non-economically profitable in large scale productions due to the needs in use ATP. Conclusion. The survey research demonstrated the effect of technological characteristics of cultivation and biotechnological properties of Saccharomyces cerevisiae on the accumulation of glutathione.

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“…There is a high industrial demand for GSH due to its favorable anti‐oxidation, anti‐aging and detoxification functions (Navarro & Obrador, 1999). GSH is mainly produced via yeast fermentation (Schmacht & Lorenz, 2017) because of the simplicity of the steps, its mild fermentation conditions, low cost of production and fast production rate (Schmacht, Lorenz, & Senz, 2017). However, GSH is an intracellular product with few extracellular penetrations and because of the influences of feedback inhibition of the intracellular GSH, the GSH content would not continue to accumulate to high concentration (Schmidt & Dringen, 2012).…”

Section: Introductionmentioning

confidence: 99%

Stimulation of in situ low intensity ultrasound on batch fermentation of Saccharomyces cerevisiae to enhance the GSH yield

Yang

Xiang

Zhang

et al. 2020

J Food Process Engineering

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In this study, an in situ 7.5 L ultrasonic fermentation tank was used to culture Saccharomyces cerevisiae at 30 C for 36 hr to explore the effects of low intensity ultrasound on the production of glutathione (GSH). Ultrasonic conditions of 28 kHz frequency, 28 W/L power density for fermentation time of 0-12 hr, 280 W/L power density for fermentation time of 12-36 hr, 3 s of interval and fixed frequency mode were used for the setup and a nonsonicated setup served as the control for the experiment. Results showed that ultrasound treatment increased total GSH production by 15.60% from a maximum value of 45.52 mg/L in the control to a maximum value of 52.62 mg/L in the sonicated sample. Also, intracellular and extracellular GSH productions in the sonicated sample increased by 5.60% and 42.92% respectively. Regular detection of the changes of biomass yield, residual sugar, dissolved oxygen and pH showed that sonication resulted in biomass increased by 7.25%, glucose consumption rate increased by 53.08% and dissolved oxygen content decreased by 41.67%. The biomass models of sonication and nonsonication treatments were established according to growth of S. cerevisiae showed that the model fitted well with Logistic regression in which the strain specific growth rate increased by 60.95% after sonication. GSH production model agreed with Luedeking-Piret regression in which GSH synthesis parameters of sonication, α and β increased by 1.96% and 19.64% in comparison to the nonsonicated sample. This study clearly demonstrated that ultrasonic stimulation could improve the biosynthesis of GSH during fermentation. Practical Applications In this study, the GSH was producted by microbial fermentation in in situ 7.5 L ultrasonic fermentation tank. During the fermentation, sonication was applied to increasing yield of GSH. The results show that the study provides a theoretical basis for manufacture GSH combined with sonication industrialization. The fermentation kinetic model was established, which provided experimental basis for continous fermentation.

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Medium optimization based on yeast's elemental composition for glutathione production in Saccharomyces cerevisiae

Cited by 14 publications

References 45 publications

Bioprocess optimization of glutathione production by Saccharomyces boulardii: biochemical characterization of glutathione peroxidase

Bioprocess optimization of glutathione production by Saccharomyces boulardii: biochemical characterization of glutathione peroxidase

Yeasts as a Glutathione Producer

Stimulation of in situ low intensity ultrasound on batch fermentation of Saccharomyces cerevisiae to enhance the GSH yield

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