Purification and Some Properties of Alcohol Acetyltransferase from Sake Yeast

Akita, Osamu; Suzuki, Syuzi; Obata, Takaji; Hara, Shodo

doi:10.1080/00021369.1990.10870152

Cited by 5 publications

(10 citation statements)

References 3 publications

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“…These results indicate that the enzyme is relatively stable below 10C and that low temperature fermentation is better for the improvement of the quality of cider. These results of thermal characteristics are similar to the enzymes isolated from brewer's yeast, sake yeast and cucumis melons (Akita et al. 1990; Minetoki et al.…”

supporting

confidence: 80%

“…2004). After detergent solubilization from the phospholipid layers, AAT is labile and difficult to be purified because this enzyme is sensitive to temperature and easy to denature because of oxidation (Yoshioka and Hashimoto 1981; Akita et al. 1990; Malcorps and Dufour 1992; Longacre et al.…”

Section: Introductionmentioning

confidence: 99%

“…AAT was first purified by gel filtration and chromatography on DEAE by Yoshioka and Hashimoto (1981). Later, Akita et al. (1990) reported the purification and properties of a membrane‐bound AAT from sake yeast.…”

Section: Introductionmentioning

confidence: 99%

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ENZYMATIC CHARACTERISTICS OF ALCOHOL ACETYLTRANSFERASE FROMHANSENIASPORA VALBYENSIS, A NON-SACCHAROMYCESYEAST

Wang

Xu²,

Фан

et al. 2008

Journal of Food Biochemistry

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Alcohol acetyltransferase (AAT) catalyzing 2‐phenylethyl alcohol and acetyl co‐enzyme A (acetyl‐CoA) to form phenylethyl acetate in Hanseniaspora valbyensis was studied. The enzyme was solubilized from lipid particles using 1% Triton X‐100 and then purified by three steps of chromatographic separations with diethylaminoethlyl (DEAE) Sepharose, Sephadex G‐75 and Octyl Sepharose, respectively. Its molecular weight was estimated to be about 37 kDa. It was discovered that AAT was most active at pH 7.0 and 30C and was stable between pH 7.0–8.0. But most activity was lost at temperatures above 10C. It was found that AAT was strongly inhibited by heavy metal ions such as Hg2+, Zn2+and Pb2+, slightly stimulated by Mg2+and slightly inhibited by ethylenediamine tetraacetic acid (EDTA) and Mn2+. The Km was 12.7 mM for 2‐phenylethyl alcohol and 23.2 mM for isoamyl alcohol. Finally, the enzyme was observed to catalyze the esterification of other alcohols with acetyl‐CoA, but have preference for 2‐phenylethyl alcohol. PRACTICAL APPLICATIONS Aroma and flavor are affected by the esters that exist in cider. Alcohol acetyltransferase (AAT) can catalyze the formation of esters. Moreover, Saccharomyces cerevisiae and non‐Saccharomyces yeast produce diverse esters with different concentration during fermentation. The influence of non‐Saccharomyces yeast on the sensory quality of wine and mixed fermentation using different yeasts have been discussed. So, the purification of AAT from Hanseniaspora valbyensis provides an understanding of the formation of esters in non‐Saccharomyces yeast.

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supporting

confidence: 80%

Section: Introductionmentioning

confidence: 99%

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ENZYMATIC CHARACTERISTICS OF ALCOHOL ACETYLTRANSFERASE FROMHANSENIASPORA VALBYENSIS, A NON-SACCHAROMYCESYEAST

Wang

Xu²,

Фан

et al. 2008

Journal of Food Biochemistry

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“…, which are much lower than the activity of Atf1 128 . Before any process conditions are optimized for higher product titers, this issue has to be addressed, e.g.…”

mentioning

confidence: 81%

“…In literature, an activity of 0.3 U/mg was reported for AtfA 127 and 3*10 -3 U/mg for Eeb1 93 , indicating that the activity of the alcoholacyltransferases is intrinsically low. Atf1 was reported to have an activity up to 190.4 U/mg 128 . Potentially protein engineering of AtfA/Eeb1 could yield a protein that has a higher activity, such as Atf1.…”

Section: 431mentioning

confidence: 99%

Production of medium-chain, a, omega-bifunctional monomers from fatty acids and n-alkanes

Nuland¹

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Saccharomyces cerevisiae Atf1p is an alcohol acetyltransferase and a thioesterase in vitro

Nancolas

Bull

Stenner

et al. 2017

Yeast

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The alcohol‐O‐acyltransferases are bisubstrate enzymes that catalyse the transfer of acyl chains from an acyl‐coenzyme A (CoA) donor to an acceptor alcohol. In the industrial yeast Saccharomyces cerevisiae this reaction produces acyl esters that are an important influence on the flavour of fermented beverages and foods. There is also a growing interest in using acyltransferases to produce bulk quantities of acyl esters in engineered microbial cell factories. However, the structure and function of the alcohol‐O‐acyltransferases remain only partly understood. Here, we recombinantly express, purify and characterize Atf1p, the major alcohol acetyltransferase from S. cerevisiae. We find that Atf1p is promiscuous with regard to the alcohol cosubstrate but that the acyltransfer activity is specific for acetyl‐CoA. Additionally, we find that Atf1p is an efficient thioesterase in vitro with specificity towards medium‐chain‐length acyl‐CoAs. Unexpectedly, we also find that mutating the supposed catalytic histidine (H191) within the conserved HXXXDG active site motif only moderately reduces the thioesterase activity of Atf1p. Our results imply a role for Atf1p in CoA homeostasis and suggest that engineering Atf1p to reduce the thioesterase activity could improve product yields of acetate esters from cellular factories. © 2017 The Authors. Yeast published by John Wiley & Sons, Ltd.

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Purification and Some Properties of Alcohol Acetyltransferase from Sake Yeast

Cited by 5 publications

References 3 publications

ENZYMATIC CHARACTERISTICS OF ALCOHOL ACETYLTRANSFERASE FROMHANSENIASPORA VALBYENSIS, A NON-SACCHAROMYCESYEAST

ENZYMATIC CHARACTERISTICS OF ALCOHOL ACETYLTRANSFERASE FROMHANSENIASPORA VALBYENSIS, A NON-SACCHAROMYCESYEAST

Production of medium-chain, a, omega-bifunctional monomers from fatty acids and n-alkanes

Saccharomyces cerevisiae Atf1p is an alcohol acetyltransferase and a thioesterase in vitro

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