Purification and some properties of alcohol acetyltransferase from sake yeast.

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

doi:10.1271/bbb1961.54.1485

Cited by 10 publications

(16 citation statements)

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“…CM-AAT1 protein was active over the pH range 6-8 consistent with the previous studies on banana [26], strawberry [27], melon [8] and yeast [28]. Na + and Mg 2+ stimulated AAT activity by 100% and 150%, respectively.…”

Section: Effect Of Ph and Various Effectors On Recombinant Cm-aat1supporting

confidence: 90%

“…These data are different from those obtained with banana AAT [26] where 10 )3 to 10 )1 M NaCl and MgCl 2 had an inhibitory effect. Akita et al [28] reported a slight effect of Mg 2+ (10% increase) in sake yeast AAT activity. However, excess Mg 2+ , but not excess Na + , caused a decrease in AAT activity.…”

Section: Effect Of Ph and Various Effectors On Recombinant Cm-aat1mentioning

confidence: 98%

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Molecular and biochemical characteristics of a gene encoding an alcohol acyl‐transferase involved in the generation of aroma volatile esters during melon ripening

Yahyaoui

Wongs‐Aree

Latché

et al. 2002

European Journal of Biochemistry

151

138

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Two genes (CM-AAT1 and CM-AAT2) with strong sequence homology (87% identity at the protein level) putatively involved in the formation of aroma volatile esters have been isolated from Charentais melon fruit. They belong to a large and highly divergent family of multifunctional plant acyl-transferases and show at most 21% identity to the only other fruit acyl-transferase characterized so far in strawberry. RT-PCR studies indicated that both genes were specifically expressed in fruit at increasing rates in the early and mid phases of ripening. Expression was severely reduced in ethylene-suppressed antisense ACC oxidase (AS) fruit and in wild-type (WT) fruit treated with the ethylene antagonist 1-MCP. Cloning of the two genes in yeast revealed that the CM-AAT1 protein exhibited alcohol acyl-transferase activity while no such activity could be detected for CM-AAT2 despite the strong homology between the two sequences. CM-AAT1 was capable of producing esters from a wide range of combinations of alcohols and acyl-CoAs. The higher the carbon chain of aliphatic alcohols, the higher the activity. Branched alcohols were esterified at differential rates depending on the position of the methyl group and the nature of the acyl donor. Phenyl and benzoyl alcohols were also good substrates, but activity varied with the position and size of the aromatic residue. The cis/trans configuration influenced activity either positively (2-hexenol) or negatively (3-hexenol). Because ripening melons evolve the whole range of esters generated by the recombinant CM-AAT1 protein, we conclude that CM-AAT1 plays a major role in aroma volatiles formation in the melon.

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Section: Effect Of Ph and Various Effectors On Recombinant Cm-aat1supporting

confidence: 90%

Section: Effect Of Ph and Various Effectors On Recombinant Cm-aat1mentioning

confidence: 98%

Molecular and biochemical characteristics of a gene encoding an alcohol acyl‐transferase involved in the generation of aroma volatile esters during melon ripening

Yahyaoui

Wongs‐Aree

Latché

et al. 2002

European Journal of Biochemistry

151

138

View full text Add to dashboard Cite

show abstract

“…On the other hand, this value is bigger than that of the Cladosporium cladosporioides No. 9 enzyme (22 kDa) (Yamakawa et al 1978) and the sake yeast enzyme (20 kDa) (Akita et al 1990). Furthermore, it is interesting to note that the molecular weight of AAT varies more widely…”

Section: Purification Of Aat From H Valbyensismentioning

confidence: 99%

“…Although some researchers discovered that the AAT from S. cerevisiae was a membrane-associated enzyme (Minetoki et al 1993;Fujii et al 1994;Lyness et al 1997), others have revealed that it is localized in lipid particles (Verstrepen et al 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 1997). AAT was first purified by gel filtration and chromatography on DEAE by Yoshioka and Hashimoto (1981).…”

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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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 10 publications

References 0 publications

Molecular and biochemical characteristics of a gene encoding an alcohol acyl‐transferase involved in the generation of aroma volatile esters during melon ripening

Molecular and biochemical characteristics of a gene encoding an alcohol acyl‐transferase involved in the generation of aroma volatile esters during melon ripening

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

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

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