Molecular cloning and nucleotide sequence of the isoamyl acetate-hydrolyzing esterase gene (EST2) from Saccharomyces cerevisiae

Fukuda, Katsuhiko; Kuwahata, Osamu; Kiyokawa, Yoshifumi; Yanagiuchi, Toshiyasu; Wakai, Yoshinori; Kitamoto, Katsuhiko; Inoue, Yoshiharu; Kimura, Akira

doi:10.1016/0922-338x(96)89447-5

Cited by 35 publications

(24 citation statements)

References 40 publications

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“…Whether these differences derive from post-translational modifications of the same protein or from the expression of different genes remains to be studied. Different genes encoding for enzymes acting on carboxylic ester bonds have been found in oenological yeast strains 6,26 and the results presented here indicate that the polymorphism of yeast esterases could be larger than previously believed.…”

Section: (Ixigxmsr Sj Iwxivewi Egxmzmx] Sr Ipigxvs Tlsvixmg Kipwmentioning

confidence: 49%

“…Formation and breakdown of specific aroma compounds have been shown to result from the action of yeast enzymes 4,6,8 . In the case of compounds that cannot be taken into the cell, these enzymes would come into contact with their specific substrates only after an active or post-mortem release into the medium.…”

Section: -2863(9'8-32mentioning

confidence: 99%

“…Esterases are expressed in yeast strains used for alcoholic beverage production and their involvement in affecting the aroma of such beverages has been the subject of several studies. By regulating the equilibrium between esters and free acids, esterases might determine ester synthesis 3,12,20,23,25 , but seem to be mainly involved in their breakdown 6,8,12 . For example, isoamyl acetate, which is thought to be synthesized from isoamyl alcohol and acetyl CoA by yeast alcohol acetyltransferase, is hydrolyzed by yeast esterases in sake mash 7,8 thus affecting the flavour quality of this beverage.…”

Section: -2863(9'8-32mentioning

confidence: 99%

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Solubilization and Activity Detection in Polyacrylamide Gels of a Membrane-Bound Esterase from an Oenological Strain of Saccharomyces cerevisiae

Rizzi

Lomolino

Lante

et al. 2003

Journal of the Institute of Brewing

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Due to the importance of esters in determining wine aroma, the presence of different esterases in soluble and insoluble cell fractions of an oenological strain of Saccharomyces cerevisiae was studied. Cells of an oenological yeast strain were separated into three fractions corresponding to the cytoplasm, periplasm and plasma membrane, all showing esterase activity. The conditions for optimal solubilization of the membrane-bound esterase were found, as well as those allowing its electrophoretic analysis, using the detergent disodium-n-lauryl-β-iminodipropionate (Deriphat) in the cathodic buffer. Comparison of the activity bands detected on gels revealed the presence of different esterase isoforms in the three sub-cellular fractions. The method can also be used as the first electrophoretic step in two-dimensional PAGE. Therefore, the procedures described are also a promising tool for the study of the yeast enzymes in relation to their effects during winemaking.

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Section: (Ixigxmsr Sj Iwxivewi Egxmzmx] Sr Ipigxvs Tlsvixmg Kipwmentioning

confidence: 49%

Section: -2863(9'8-32mentioning

confidence: 99%

Section: -2863(9'8-32mentioning

confidence: 99%

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Solubilization and Activity Detection in Polyacrylamide Gels of a Membrane-Bound Esterase from an Oenological Strain of Saccharomyces cerevisiae

Rizzi

Lomolino

Lante

et al. 2003

Journal of the Institute of Brewing

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“…As esterases are responsible for the breakdown of esters, it is clear that their activity may affect the concentration of certain volatile esters. In S. cerevisiae, for example, it has been shown that the balance between ester-synthesizing enzymes and esterases, such as Iah1p (also known as Est2p), is important for the net rate of ester accumulation (27)(28)(29)(30) (59,60,61,62,70). This indicates that esterases may also play a role in the formation of certain esters (42,62,70).…”

Section: Discussionmentioning

confidence: 99%

“…However, the effect of Iah1p on other esters, such as the group of ethyl esters, has not yet been thoroughly studied. In addition, partial purification of esterase activities in S. cerevisiae has shown that at least two different fractions with esterase activity exists, indicating that the proteome may contain more, as-yet-unidentified esterases (28,33,59,60,70,73,82). Hence, a possible role for these additional esterases in ester synthesis in brewer's yeast cannot be ruled out.…”

Section: Discussionmentioning

confidence: 99%

Expression Levels of the Yeast Alcohol Acetyltransferase GenesATF1,Lg-ATF1, andATF2Control the Formation of a Broad Range of Volatile Esters

Verstrepen

Laere

Vanderhaegen

et al. 2003

Appl Environ Microbiol

336

322

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Volatile aroma-active esters are responsible for the fruity character of fermented alcoholic beverages such as beer and wine. Esters are produced by fermenting yeast cells in an enzyme-catalyzed intracellular reaction. In order to investigate and compare the roles of the known Saccharomyces cerevisiae alcohol acetyltransferases, Atf1p, Atf2p and Lg-Atf1p, in volatile ester production, the respective genes were either deleted or overexpressed in a laboratory strain and a commercial brewing strain. Subsequently, the ester formation of the transformants was monitored by headspace gas chromatography and gas chromatography combined with mass spectroscopy (GC-MS). Analysis of the fermentation products confirmed that the expression levels of ATF1 and ATF2 greatly affect the production of ethyl acetate and isoamyl acetate. GC-MS analysis revealed that Atf1p and Atf2p are also responsible for the formation of a broad range of less volatile esters, such as propyl acetate, isobutyl acetate, pentyl acetate, hexyl acetate, heptyl acetate, octyl acetate, and phenyl ethyl acetate. With respect to the esters analyzed in this study, Atf2p seemed to play only a minor role compared to Atf1p. The atf1⌬ atf2⌬ double deletion strain did not form any isoamyl acetate, showing that together, Atf1p and Atf2p are responsible for the total cellular isoamyl alcohol acetyltransferase activity. However, the double deletion strain still produced considerable amounts of certain other esters, such as ethyl acetate (50% of the wild-type strain), propyl acetate (50%), and isobutyl acetate (40%), which provides evidence for the existence of additional, as-yet-unknown ester synthases in the yeast proteome. Interestingly, overexpression of different alleles of ATF1 and ATF2 led to different ester production rates, indicating that differences in the aroma profiles of yeast strains may be partially due to mutations in their ATF genes.During fermentation processes, yeast cells produce a broad range of aroma-active substances which greatly affect the complex flavor of fermented alcoholic beverages. While these secondary metabolites are often formed only in trace amounts, their concentrations determine the distinct aroma of these beverages. Flavor-active substances produced by fermenting yeast cells can be divided into five main groups: sulfur-containing molecules, organic acids, higher alcohols, carbonyl compounds, and volatile esters (32,39,56,57,66). Of these categories, volatile esters represent the largest and most important group. They are responsible for the highly desired fruity character of beer and, to a lesser extent, other alcoholic beverages, such as wine.The major flavor-active esters in beer are acetate esters such as ethyl acetate (solvent-like aroma), isoamyl acetate (banana flavor), and phenylethyl acetate (flowery, rose aroma). In addition, C 6 -C 10 medium-chain fatty acid ethyl esters such as ethyl hexanoate (ethyl caproate) and ethyl octanoate (ethyl caprylate), which have "sour apple" aromas, are also important for the overall bouqu...

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A novel esterase fromSaccharomyces carlsbergensis, a possible function for the yeastTIP1 gene

Horsted

Dey

Holmberg

et al. 1998

Yeast

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Molecular cloning and nucleotide sequence of the isoamyl acetate-hydrolyzing esterase gene (EST2) from Saccharomyces cerevisiae

Cited by 35 publications

References 40 publications

Solubilization and Activity Detection in Polyacrylamide Gels of a Membrane-Bound Esterase from an Oenological Strain of Saccharomyces cerevisiae

Solubilization and Activity Detection in Polyacrylamide Gels of a Membrane-Bound Esterase from an Oenological Strain of Saccharomyces cerevisiae

Expression Levels of the Yeast Alcohol Acetyltransferase GenesATF1,Lg-ATF1, andATF2Control the Formation of a Broad Range of Volatile Esters

A novel esterase fromSaccharomyces carlsbergensis, a possible function for the yeastTIP1 gene

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