Engineering redox cofactor utilization for detoxification of glycolaldehyde, a key inhibitor of bioethanol production, in yeast Saccharomyces cerevisiae

Jayakody, Lahiru N.; Horie, Kenta; Hayashi, Naoaki; Kitagaki, Hiroshi

doi:10.1007/s00253-013-4997-4

Cited by 32 publications

(15 citation statements)

References 41 publications

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“…To investigate the time course of yeast growth in the presence of various GlcCers, the OD 600 of independent duplicate cultures was measured. The maximum specific growth rate ( max ) was calculated according to the method described previously (23)(24)(25)(26).…”

Section: Methodsmentioning

confidence: 99%

Glucosylceramide Contained in Koji Mold-Cultured Cereal Confers Membrane and Flavor Modification and Stress Tolerance to Saccharomyces cerevisiae during Coculture Fermentation

Sawada

Sato

Hamajima

et al. 2015

Appl Environ Microbiol

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fIn nature, different microorganisms create communities through their physiochemical and metabolic interactions. Many fermenting microbes, such as yeasts, lactic acid bacteria, and acetic acid bacteria, secrete acidic substances and grow faster at acidic pH values. However, on the surface of cereals, the pH is neutral to alkaline. Therefore, in order to grow on cereals, microbes must adapt to the alkaline environment at the initial stage of colonization; such adaptations are also crucial for industrial fermentation. Here, we show that the yeast Saccharomyces cerevisiae, which is incapable of synthesizing glucosylceramide (GlcCer), adapted to alkaline conditions after exposure to GlcCer from koji cereal cultured with Aspergillus kawachii. We also show that various species of GlcCer derived from different plants and fungi similarly conferred alkali tolerance to yeast. Although exogenous ceramide also enhanced the alkali tolerance of yeast, no discernible degradation of GlcCer to ceramide was observed in the yeast culture, suggesting that exogenous GlcCer itself exerted the activity. Exogenous GlcCer also increased ethanol tolerance and modified the flavor profile of the yeast cells by altering the membrane properties. These results indicate that GlcCer from A. kawachii modifies the physiology of the yeast S. cerevisiae and demonstrate a new mechanism for cooperation between microbes in food fermentation. In nature, microbial communities are formed through metabolic and physiochemical interactions among different microorganisms (1). The pH of cereals that have ripened and dropped from the husk to the ground is neutral to alkaline (2-4). Fermentation microbes, including Saccharomyces cerevisiae, Lactobacillus lactis, and Acetobacter aceti, efficiently utilize carbohydrates in the cereals to produce organic acids (1). As a result, the microbial communities that colonize such carbohydrate-rich cereals, as well as those found in fermented foods, reduce the pH to 4.0 to 6.0. After these microbes establish an acidic pH, they can dominate the environment because they can grow faster at acidic pH values than other microbes. Therefore, these microbes must first adapt to the alkaline environment at the initial stage of colonization on cereal; such adaptation mechanisms are also crucial for industrial fermentation.Traditional alcoholic beverages are produced via the fermentation of cereals by the yeast S. cerevisiae, which can efficiently catabolize glucose to produce ethanol. However, because S. cerevisiae is incapable of hydrolyzing starch to glucose, additional biological catalysts are often added to the fermentation reaction. For example, malt catalyzes starch hydrolysis in wheat-and barleyderived beverages such as whiskey and beer. In East and Southeast Asia, various cereals fermented by fungi are widely used as starchhydrolyzing catalysts for the production of rice-derived alcoholic beverages. These include Japanese sake (5, 6) and shochu, Korean Makgeolli, and Chinese Huangjiu, as well as various other fermented foods ...

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Section: Methodsmentioning

confidence: 99%

Glucosylceramide Contained in Koji Mold-Cultured Cereal Confers Membrane and Flavor Modification and Stress Tolerance to Saccharomyces cerevisiae during Coculture Fermentation

Sawada

Sato

Hamajima

et al. 2015

Appl Environ Microbiol

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“…In addition, a variety of inhibitory compounds are released, such as furfural, hydroxymethylfurfural, methylglyoxal, and acetate (4)(5)(6). It is therefore desirable not only to expand the substrate range of S. cerevisiae (3,(7)(8)(9) but also to increase its inhibitor tolerance (10)(11)(12)(13).…”

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confidence: 99%

Increasing Anaerobic Acetate Consumption and Ethanol Yields in Saccharomyces cerevisiae with NADPH-Specific Alcohol Dehydrogenase

Henningsen

Hon

Covalla

et al. 2015

Appl Environ Microbiol

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Saccharomyces cerevisiae has recently been engineered to use acetate, a primary inhibitor in lignocellulosic hydrolysates, as a cosubstrate during anaerobic ethanolic fermentation. However, the original metabolic pathway devised to convert acetate to ethanol uses NADH-specific acetylating acetaldehyde dehydrogenase and alcohol dehydrogenase and quickly becomes constrained by limited NADH availability, even when glycerol formation is abolished. We present alcohol dehydrogenase as a novel target for anaerobic redox engineering of S. cerevisiae. Introduction of an NADPH-specific alcohol dehydrogenase (NADPH-ADH) not only reduces the NADH demand of the acetate-to-ethanol pathway but also allows the cell to effectively exchange NADPH for NADH during sugar fermentation. Unlike NADH, NADPH can be freely generated under anoxic conditions, via the oxidative pentose phosphate pathway. We show that an industrial bioethanol strain engineered with the original pathway (expressing acetylating acetaldehyde dehydrogenase from Bifidobacterium adolescentis and with deletions of glycerol-3-phosphate dehydrogenase genes GPD1 and GPD2) consumed 1.9 g liter ؊1 acetate during fermentation of 114 g liter ؊1 glucose. Combined with a decrease in glycerol production from 4.0 to 0.1 g liter ؊1 , this increased the ethanol yield by 4% over that for the wild type. We provide evidence that acetate consumption in this strain is indeed limited by NADH availability. By introducing an NADPH-ADH from Entamoeba histolytica and with overexpression of ACS2 and ZWF1, we increased acetate consumption to 5.3 g liter ؊1and raised the ethanol yield to 7% above the wild-type level. Saccharomyces cerevisiae is the principal microorganism used to produce ethanol, of which 93 billion liters were globally produced in 2014 (1). However, new strains are needed for the production of second-generation biofuels. Pretreatment and monomerization of lignocellulosic substrates produce complex sugar mixtures, including the C 5 sugars xylose and arabinose that are poorly fermented by most wild-type S. cerevisiae strains (2, 3). In addition, a variety of inhibitory compounds are released, such as furfural, hydroxymethylfurfural, methylglyoxal, and acetate (4-6). It is therefore desirable not only to expand the substrate range of S. cerevisiae (3, 7-9) but also to increase its inhibitor tolerance (10-13).Acetate is released during deacylation of hemicellulose and lignin and can be present in concentrations of Ͼ10 g liter Ϫ1 in cellulosic hydrolysates (4,14). At low pH particularly, this weak organic acid is a potent inhibitor of S. cerevisiae metabolism (15-17). Unfortunately, wild-type S. cerevisiae strains are poorly equipped to eliminate acetate from the medium under anoxic conditions. S. cerevisiae can grow on acetate as the sole carbon source under oxic conditions by converting acetate to acetyl coenzyme A (acetyl-CoA) with acetyl-CoA synthetase (ACS) (EC 6.2.1.1) and by either respiring acetyl-CoA in the tricarboxylic acid (TCA) cycle or upgrading acetyl-CoA to four-...

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“…This would accelerate the glycolytic flux and enhance the NADH level in the central metabolic pathway [15]. At the same time, NADH-dependent alcohol dehydrogenase (ADH) activity might increase, causing up-regulation of acetaldehyde to ethanol conversion, accompanied by the oxidation of NADH to NAD + [36,37] (figure 1). Therefore, the NADH level obtained from 1 min exposure was decreased compared with the control because of the lower level of ATP ( figure 4 1 min versus figure 3 1 min).…”

Section: Discussionmentioning

confidence: 99%

A novel approach to regulate cell membrane permeability for ATP and NADH formation inSaccharomyces cerevisiaeinduced by air cold plasma

Dong

Liu

Xiong

2017

Plasma Sci. Technol.

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Air cold plasma has been used as a novel method for enhancing microbial fermentation. The aim of this work was to explore the effect of plasma on membrane permeability and the formation of ATP and NADH in Saccharomyces cerevisiae, so as to provide valuable information for largescale application ofplasma in the fermentation industry. Suspensions of S. cerevisiae cells were exposed to air cold plasma for 0, 1, 2, 3, 4 and 5 min, and then subjected to various analyses prior to fermentation (0 h) and at the 9 and 21 h stages of fermentation. Compared with nonexposed cells, cells exposed to plasma for 1 min exhibited a marked increase in cytoplasmic free Ca 2+ concentration as a result of the significant increase in membrane potential prior to fermentation. At the same time, the ATP level in the cell suspension decreased by about 40%, resulting in a reduction ofabout 60% in NADH prior to culturing. However, the levels of ATP and NADH in the culture at the 9 and 21 h fermentation stages were different from the level at 0 h. Taken together, the results indicated that exposure of S. cerevisiae to air cold plasma could increase its cytoplasmic free Ca 2+ concentration by improving the cell membrane potential, consequently leading to changes in ATP and NADH levels.

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Engineering redox cofactor utilization for detoxification of glycolaldehyde, a key inhibitor of bioethanol production, in yeast Saccharomyces cerevisiae

Cited by 32 publications

References 41 publications

Glucosylceramide Contained in Koji Mold-Cultured Cereal Confers Membrane and Flavor Modification and Stress Tolerance to Saccharomyces cerevisiae during Coculture Fermentation

Glucosylceramide Contained in Koji Mold-Cultured Cereal Confers Membrane and Flavor Modification and Stress Tolerance to Saccharomyces cerevisiae during Coculture Fermentation

Increasing Anaerobic Acetate Consumption and Ethanol Yields in Saccharomyces cerevisiae with NADPH-Specific Alcohol Dehydrogenase

A novel approach to regulate cell membrane permeability for ATP and NADH formation inSaccharomyces cerevisiaeinduced by air cold plasma

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