Cofactor engineering in Saccharomyces cerevisiae: Expression of a H2O-forming NADH oxidase and impact on redox metabolism

Heux, Stéphanie; Cachon, Rémy; Dequin, Sylvie

doi:10.1016/j.ymben.2005.12.003

Cited by 141 publications

(118 citation statements)

References 49 publications

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“…Consequently, numerous attempts have been made to engineer Saccharomyces cerevisiae yeast strains with reduced ethanol yields, which would offer faster and less expensive biological alternatives to the current physical processes available for the production of low-and reduced-alcohol wines (29). The biological approaches used so far are all based on diverting sugar metabolism toward byproducts other than ethanol by metabolic engineering strategies (7,8,18,19,21,22,30). However, these strategies have so far not satisfied the need to obtain a significant reduction in the ethanol yield without causing the accumulation of undesirable secondary products and/or without affecting yeast physiology.…”

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confidence: 99%

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Ehsani

Fernández

Biosca

et al. 2009

Appl Environ Microbiol

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110

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Engineered Saccharomyces cerevisiae strains overexpressing GPD1, which codes for glycerol-3-phosphate dehydrogenase, and lacking the acetaldehyde dehydrogenase Ald6 display large-scale diversion of the carbon flux from ethanol toward glycerol without accumulating acetate. Although GPD1 ald6 strains have great potential for reducing the ethanol contents in wines, one major side effect is the accumulation of acetoin, having a negative sensory impact on wine. Acetoin is reduced to 2,3-butanediol by the NADH-dependent 2,3-butanediol dehydrogenase Bdh1. In order to investigate the influence of potential factors limiting this reaction, we overexpressed BDH1, coding for native NADH-dependent Bdh1, and the engineered gene BDH1 221,222,223 , coding for an NADPH-dependent Bdh1 enzyme with the amino acid changes 221 EIA 223 to 221 SRS 223, in a glycerol-overproducing wine yeast. We have shown that both the amount of Bdh1 and the NADH availability limit the 2,3-butanediol dehydrogenase reaction. During wine fermentation, however, the major limiting factor was the level of synthesis of Bdh1. Consistent with this finding, the overproduction of native or engineered Bdh1 made it possible to redirect 85 to 90% of the accumulated acetoin into 2,3-butanediol, a compound with neutral sensory characteristics. In addition, the production of diacetyl, a compound causing off-flavor in alcoholic beverages, whose production is increased in glycerol-overproducing yeast cells, was decreased by half. The production of higher alcohols and esters, which was slightly decreased or unchanged in GPD1 ald6 cells compared to that in the control cells, was not further modified in BDH1 cells. Overall, rerouting carbons toward glycerol and 2,3-butanediol represents a new milestone in the engineering of a low-alcohol yeast with desirable organoleptic features, permitting the decrease of the ethanol contents in wines by up to 3°.

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confidence: 99%

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Ehsani

Fernández

Biosca

et al. 2009

Appl Environ Microbiol

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110

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“…XDH enzyme activity was determined as previously described (4, 30) using a CECIL 2000 series (Bioquest, England) spectrophotometer at 340 nm. The enzyme activity of water-forming NADH oxidase was measured as micromoles of converted substrate per milligram of protein per minute according to Heux et al (10).…”

Section: Methodsmentioning

confidence: 99%

“…2A and B). The slow growth and xylose consumption rates probably were caused by the overexpression of noxE, which might lead to redox imbalance, hence affecting cell growth (10). To clarify this phenomenon, the enzyme activities of NADH oxidase and XDH were measured to see if the microcycle we are trying to build was in balance.…”

Section: Oxygen-limited Batch Fermentation Of Kam-3x Carrying Noxementioning

confidence: 99%

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Decreased Xylitol Formation during Xylose Fermentation in Saccharomyces cerevisiae Due to Overexpression of Water-Forming NADH Oxidase

Zhang

Liu

Ding

2012

Appl Environ Microbiol

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The recombinant xylose-fermenting Saccharomyces cerevisiae strain harboring xylose reductase (XR) and xylitol dehydrogenase (XDH) from Scheffersomyces stipitis requires NADPH and NAD ؉ , creates cofactor imbalance, and causes xylitol accumulation during growth on D-xylose. To solve this problem, noxE, encoding a water-forming NADH oxidase from Lactococcus lactis driven by the PGK1 promoter, was introduced into the xylose-utilizing yeast strain KAM-3X. A cofactor microcycle was set up between the utilization of NAD ؉ by XDH and the formation of NAD ؉ by water-forming NADH oxidase. Overexpression of noxE significantly decreased xylitol formation and increased final ethanol production during xylose fermentation. Under xylose fermentation conditions with an initial D-xylose concentration of 50 g/liter, the xylitol yields for of KAM-3X(pPGK1-noxE) and control strain KAM-3X were 0.058 g/g xylose and 0.191 g/g, respectively, which showed a 69.63% decrease owing to noxE overexpression; the ethanol yields were 0.294 g/g for KAM-3X(pPGK1-noxE) and 0.211 g/g for the control strain KAM-3X, which indicated a 39.33% increase due to noxE overexpression. At the same time, the glycerol yield also was reduced by 53.85% on account of the decrease in the NADH pool caused by overexpression of noxE.

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“…Yeast is the key raw material in many consumer products, for instance, in the production of beverages (Heux et al, 2006), antibiotics (Kim, 2004) and recombinant proteins (Kato et al, 1998). In addition, it plays an essential role in the energy sector for the production of biomass or biofuel.…”

Section: Introductionmentioning

confidence: 99%

Intensification of yeast production with microbubbles

Hanotu

Kong

Zimmerman

2016

Food and Bioproducts Processing

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Cofactor engineering in Saccharomyces cerevisiae: Expression of a H2O-forming NADH oxidase and impact on redox metabolism

Cited by 141 publications

References 49 publications

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Engineering of 2,3-Butanediol Dehydrogenase To Reduce Acetoin Formation by Glycerol-Overproducing, Low-AlcoholSaccharomyces cerevisiae

Decreased Xylitol Formation during Xylose Fermentation in Saccharomyces cerevisiae Due to Overexpression of Water-Forming NADH Oxidase

Intensification of yeast production with microbubbles

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