Importance of the structural zinc atom for the stability of yeast alcohol dehydrogenase

Magonet, E.; P, Hayen; Delforge, Dominique; Delaive, Edouard; Remacle, José

doi:10.1042/bj2870361

Cited by 126 publications

(91 citation statements)

References 20 publications

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“…Yeast cellular zinc content therefore appears to directly influence ethanol production. Despite the fact that alcohol dehydrogenase ADH (the terminal enzyme in ethanol fermentation) is a zinc metalloenzyme 20 , yeast cells with high zinc content produced low concentrations of ethanol. This could be related to zinc accumulation in vacuoles 18,19,22,8 not made available to ADH or to the presence in the studied strains of other zinc enzymes with higher affinities for Zn.…”

Section: Effects Of Zn-preconditioned Yeastmentioning

confidence: 99%

“…This metal is used as cofactor in numerous enzymes 32 and plays a structural and functional role in proteins and nucleic acids 1,26,31 . For yeast fermentation processes, zinc is absolutely essential for alcohol production, as it is required for the functioning of the terminal enzymatic step in ethanologenesis, namely alcohol dehydrogenase 20 . In some industrial yeast-based processes, such as wine-making, zinc concentrations in the medium (e.g.…”

Section: Introductionmentioning

confidence: 99%

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Influence of Zinc on Distiller's Yeast: Cellular Accumulation of Zinc and Impact on Spirit Congeners

Nicola

Hall

Melville

et al. 2009

Journal of the Institute of Brewing

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Accumulation of zinc by a whisky distilling yeast strain of Saccharomyces cerevisiae was studied during fermentation of malt wort and synthetic defined medium. Zinc uptake by yeast cells was very rapid in malt wort, as zinc (0.32 μg/mL) was completely removed from the fermentation medium within one hour. The type of fermentable carbohydrate had an impact on the kinetics of zinc accumulation, with maltose most effective at enhancing metal uptake at zinc concentrations above 3.2 µg/mL. Enriching yeast cells with zinc by "preconditioning" impacted on the production of flavour congeners in the distillates produced from fermented cultures. Such distillates were characterised by an altered flavour and aroma profile. In particular, the production of some higher alcohols increased when yeast cells were preconditioned with zinc. This phenomenon is yeast strain related. Industrial fermentation processes, including brewing and distilling, may benefit from optimization of zinc bioavailability in yeast cultures resulting in more efficient fermentations and improved product quality.

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Section: Effects Of Zn-preconditioned Yeastmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Influence of Zinc on Distiller's Yeast: Cellular Accumulation of Zinc and Impact on Spirit Congeners

Nicola

Hall

Melville

et al. 2009

Journal of the Institute of Brewing

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“…With such a stabilizing role, the zinc is referred to as`structural zinc', as has been reported for other proteins including alcohol dehydrogenase [23], adenylate kinase [24] and GP32 of bacteriophage T4 [25]. In these cases, Zn is ligated by either four Cys residues or three Cys and one His.…”

Section: Discussionmentioning

confidence: 99%

Zinc and an N‐terminal extra stretch of the ferredoxin from a thermoacidophilic archaeon stabilize the molecule at high temperature

Kojoh

Matsuzawa

Wakagi

1999

European Journal of Biochemistry

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Ferredoxin from the thermoacidophilic archaeon Sulfolobus sp. strain 7 has a 36-residue extra domain at its N-terminus and a 67-residue core domain carrying two iron±sulfur clusters. A zinc ion is held at the interface of the two domains through tetrahedral coordination of three histidine residues (26, 219 and 234) and one aspartic acid residue (276) [Fujii, T., Hata, Y., Oozeki, M., Moriyama, H., Wakagi, T., Tanaka, N. & Oshima, T. (1997) Biochemistry 36, 1505±1513]. To elucidate the roles of the novel zinc ion and the extra N-terminal domain, a series of truncated mutants was constructed: G1, V12, S17, G23, L31 and V38, which lack residues 0, 11, 16, 22, 30 and 37 starting from the N-terminus, respectively. A mutant with two histidine residues each replaced by an alanine residue, H16A/H19A, was also constructed. All the mutant ferredoxins had two iron±sulfur clusters, while zinc was retained only in G1 and V12. The thermal stability of the proteins was investigated by monitoring A 408 ; the melting temperature (T m ) was <109 8C for the natural ferredoxin, <109 8C for G1, 97.6 8C for V12, 89.0 8C for S17, 89.2 8C for G23, 89.3 8C for L31, 82.1 8C for V38, and 89.4 8C for H16A/H19A. K m and V max values of 2-oxoglutarate:ferredoxin oxidoreductase for natural ferredoxin, G1, S17 and L31 were similar, suggesting that electron-accepting activities were not affected by the deletion. The combination of CD and fluorescent spectroscopic analyses with truncated mutant S17 indicated that not only the clusters but also the secondary and tertiary structures were simultaneously degraded at a T m around 89 8C. These results unequivocally demonstrate that the zinc ion and certain parts, but not all, of the extra sequence stretch in the N-terminal domain are responsible not for function but for thermal stabilization of the molecule.Keywords: archaea; ferredoxin; stability; zinc.Ferredoxin is a small protein widely distributed among all living organisms, with one or more redox center(s) composed of acid-labile sulfur and non-heme iron participating in various oxidation±reduction processes in the cell [1,2]. Archaeal ferredoxins are not classified into a simple`archaeal type', but into different categories based on both the clusters and amino-acid sequences; the ferredoxins from thermoacidophilic archaea belong to the 7Fe-bicluster type, those from anaerobic hyperthermophilic archaea to the 4Fe-monocluster type, those from extremely halophilic archaea to the 2Fe-type, and so on [2,3]. It is, however, striking that they play a common role as electron acceptors in the CoA-dependent oxidative decarboxylation of 2-oxoacids, which is seldom found in eubacteria or eukaryotes [4]. In contrast with the widely found 2-oxoacid dehydrogenase multienzyme complex with NAD(P) as an electron acceptor [5], 2-oxoacid:ferredoxin oxidoreductase (OFOR) is a very simple enzyme comprising one to four subunits, with a total molecular mass of around 103±260 kDa [6,7].The primary structure, comprising 103 amino acid residues, of the ferredoxin from the...

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“…The alcohol dehydrogenase lyophilized powder from Saccharomyces cerevisiae (Sigma-Aldrich A7011) (ADH) was used with no further purification. This enzyme has a molecular weight 141-151 kDa, an isoelectric point between 5.4-5.8 and the optimal pH is reported to be in the range of 8.6 and 9.0 [17][18][19].…”

Section: Methodsmentioning

confidence: 99%

“…The other zinc atom (structural zinc) is linked to four cysteine residues (97, 100, 103 and 111) and it is in an external location protected by disulphide bridges. Its function is still unclear, although it seems to have an important conformational role, by stabilizing the tertiary structure of each subunit [19]. Its removal from the enzyme did not reduce the catalytic activity, but the enzyme was more susceptible to heat denaturation [19].…”

Section: Introductionmentioning

confidence: 99%

Achieving Enzyme Stability Using a Simple Fabrication Procedure: The Alcohol Dehydrogenase Example

Ribau¹,

Fortunato²

2018

JPP

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Abstract:The use of screen-printing biosensors has been updated in this article as a tool to analyze the electron transfer process involving redox proteins or enzymes. The aim of this research was to fabricate a simple apparatus which allowed the use of the enzymes (in the solid state) to maintain their stability. To prove this concept an enzyme in the solid state was mixed with the carbon ink and this mixture was used to print the working electrode. We choose as proving the alcohol dehydrogenase. The first reason is because it metabolizes the alcohol, which can be present in biological samples of blood, saliva and urine and also in the beverage; the second is that this enzyme is still a challenge to electrochemistry due to having lower stability in sensors. The results show that in this device the enzyme was active and stable during all the experiments and in the experimental conditions that could catalyze the ethanol to acetaldehyde. These devices have the advantage of being disposable, cheap and are easy to fabricate. And also, they do not need expensive tools to be fabricated, they only need 2 µL of electrolyte or sample, and they need lower amounts of enzyme to permit electrochemical studies.

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Importance of the structural zinc atom for the stability of yeast alcohol dehydrogenase

Cited by 126 publications

References 20 publications

Influence of Zinc on Distiller's Yeast: Cellular Accumulation of Zinc and Impact on Spirit Congeners

Influence of Zinc on Distiller's Yeast: Cellular Accumulation of Zinc and Impact on Spirit Congeners

Zinc and an N‐terminal extra stretch of the ferredoxin from a thermoacidophilic archaeon stabilize the molecule at high temperature

Achieving Enzyme Stability Using a Simple Fabrication Procedure: The Alcohol Dehydrogenase Example

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