Cloning and Nucleotide Sequence of the Alcohol Acetyltransferase II Gene (<i>ATF2</i>) from<i>Saccharomyces cerevisiae</i>Kyokai No. 7

Nagasawa, Naoshi; Bogaki, Takayuki; Iwamatsu, Akihiro; Hamachi, Masaaki; Kumagai, Chieko

doi:10.1271/bbb.62.1852

Cited by 97 publications

(73 citation statements)

References 17 publications

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“…Evolution of isoamyl alcohol (A) and isobutanol (B) levels during fermentation of wort brewed with 100% Scarlett or 50% Scarlett and 50% of dark malt (Melanoidin, Cara-aroma and roasted malt). Properties of these wort samples are presented in Table VII. are the key enzymes involved in the biosynthesis of acetate esters 15,25,27 . Deletion and overexpression studies indicated that the ATF1 gene is responsible for the formation of both isoamyl acetate and ethyl acetate, whereas ATF2 mainly leads to isoamyl acetate 35 .…”

Section: Fusel Alcohols and Estersmentioning

confidence: 99%

Impact of Dark Specialty Malts on Extract Composition and Wort Fermentation

Coghe

D'Hollander

Verachtert

et al. 2005

Journal of the Institute of Brewing

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Dark specialty malts are important ingredients for the production of several beer styles. These malts not only impart colour, flavour and antioxidative activity to wort and beer, they also affect the course of wort fermentations and the production of flavouractive yeast metabolites. The application of considerable levels of dark malt was found to lower the attenuation, mainly as a result of lower levels of fermentable sugars and amino acids in dark wort samples. In fact, from the darkest caramel malts and from roasted malts, practically no fermentable material can be hydrolysed by pilsner malt enzymes during mashing. Compared to wort brewed with 50% pilsner malt and 50% dark caramel malt or roasted malt, wort brewed with 100% pilsner malt contained nearly twice as much fermentable sugars and amino acids. Reduced levels of yeast nutrients also lowered the fermentation rate, ranging from 1.7°P/day for the reference pilsner wort of 9 EBC to 1.1°P/day for the darkest wort (890 EBC units), brewed with 50% roasted malt. This additionally indicates that lower attenuation values for dark wort are partially due to the inhibitory effects of Maillard compounds on yeast metabolism. The application of dark caramel or roasted malts further led to elevated levels of the vicinal diketones diacetyl and 2,3-pentanedione. Only large levels of roasted malt gave rise to two significant diacetyl peaks during fermentation. The level of ethyl acetate in beer was inversely related to colour, whereas the level of isoamyl acetate appeared to be affected by the use of roasted malt. With large levels of this malt type, negligible isoamyl acetate was generated during fermentation.

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Section: Fusel Alcohols and Estersmentioning

confidence: 99%

Impact of Dark Specialty Malts on Extract Composition and Wort Fermentation

Coghe

D'Hollander

Verachtert

et al. 2005

Journal of the Institute of Brewing

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“…In addition, little is known about the contribution of other enzymes to the total ester-synthesizing activity in fermenting yeast cells. Nagasawa et al (49) and Yoshimoto et al (83) suggested that Atf2p might be responsible for the remaining 20% of isoamyl acetate production in a atf1⌬ disruptant, but this remains to be confirmed. In addition, it remains unclear whether the Atf proteins are also responsible for the formation of other esters besides isoamyl acetate and ethyl acetate.…”

mentioning

confidence: 98%

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

345

311

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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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“…While extensive research has been carried out on the enzymes responsible for ester formation by wine strains of Saccharomyces cerevisiae (22,23,45,51), esterase activity for winerelated LAB is not well documented. Most characterization of esterases in LAB has focused on dairy isolates (9,(16)(17)(18)20).…”

mentioning

confidence: 99%

Cloning and Characterization of an Intracellular Esterase from the Wine-Associated Lactic Acid BacteriumOenococcus oeni

Sumby

Matthews

Grbin

et al. 2009

Appl Environ Microbiol

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We report the cloning and characterization of EstB28, the first esterase to be so characterized from the wine-associated lactic acid bacterium, Oenococcus oeni. The published sequence for O. oeni strain PSU-1 was used to identify putative esterase genes and design PCR primers in order to amplify the corresponding region from strain Ooeni28, an isolate intended for inoculation of wines. In this way a 912-bp open reading frame (ORF) encoding a putative esterase of 34.5 kDa was obtained. The amino acid sequence indicated that EstB28 is a member of family IV of lipolytic enzymes and contains the GDSAG motif common to other lactic acid bacteria. This ORF was cloned into Escherichia coli using an appropriate expression system, and the recombinant esterase was purified. Characterization of EstB28 revealed that the optimum temperature, pH, and ethanol concentration were 40°C, pH 5.0, and 28% (vol/vol), respectively. EstB28 also retained marked activity under conditions relevant to winemaking (10 to 20°C, pH 3.5, 14% [vol/vol] ethanol). Kinetic constants were determined for EstB28 with p-nitrophenyl (pNP)-linked substrates ranging in chain length from C 2 to C 18 . EstB28 exhibited greatest specificity for C 2 to C 4 pNP-linked substrates.

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Cloning and Nucleotide Sequence of the Alcohol Acetyltransferase II Gene (ATF2) fromSaccharomyces cerevisiaeKyokai No. 7

Cited by 97 publications

References 17 publications

Impact of Dark Specialty Malts on Extract Composition and Wort Fermentation

Impact of Dark Specialty Malts on Extract Composition and Wort Fermentation

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

Cloning and Characterization of an Intracellular Esterase from the Wine-Associated Lactic Acid BacteriumOenococcus oeni

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