Alcohol Acetyltransferase Activity Research Articles

The effects of different rootstocks on aroma components, activities and genes expression of aroma-related enzymes in oriental melon fruit.

Grafting is widely applied in the cultivation of melon. In this study, 'Qinmi No.1' (Cucumis melo L.(QG)) and 'Ribenxuesong' (Cucurbita maxima Duch. (RG)) were used as rootstocks for 'Qingxin Yangjiaocui' (Cucumis melo L.). The results showed that grafting with muskmelon rootstocks had no significant effect on fruit aroma, but grafting with pumpkin rootstocks significantly reduced the odor intensity and odor preference scores of melon fruits. Compared with the fruits from self-grafted plants (SG), four new aromatic volatiles with a sweet smell were detected, the alcohol dehydrogenase (ADH) activity was significantly decreased at 30 DAP, but unaffected at 42 DAP in QG fruits. There was no difference for alcohol acetyltransferase (AAT) activity between QG and SG fruits. The expression level of CmADH2 was significantly higher at 30 DAP and 42 DAP, but CmAAT2 was significantly lower at 42 DAP in QG fruits compared with SG fruits. In RG fruits, the main aroma compounds including butanoic acid ethyl ester, 2-methyl-2-butene-1-al, and 2-methylheptan-1-al were absent, while the volatile compounds with unpleasant odor characteristics including trans, cis-2,6-nonadien-1-ol, (E,E)-2,4-heptadienal, octanoic acid, and styrene were detected. Compared with SG fruits, 1-nonanol and 1-heptanol with green odor characteristics were significantly increased, but eucalyptol and farnesene with fruity aroma characteristics were significantly decreased in RG fruits. The ADH activity of RG fruits was significantly lower than that of SG fruits at 30 DAP and the AAT activity was significantly lower than that of SG fruits at 42 DAP. In addition, the expression levels of CmADH and CmAAT homologs in RG fruits were significantly lower than those in SG or QG fruits. These results show that grafting with pumpkin rootstocks affected the main aroma components, reduced ADH and AAT activities, and down-regulated the expression levels of CmADHs and CmAATs in the melon fruits. This study reveals the mechanism of different rootstocks on melon fruit aroma quality, and lays a theoretical foundation for the selection of rootstocks in melon production. Future studies using overexpression or CRISPR/CAS system to obtain stable transgenic lines of genes encoding key aromatic volatiles, would be promising to effectively improve the flavor quality of melon.

Screening of the Volatile Composition of Moroccan Olive Oils by Using SPME/GC-MS-FID over a Two-Year Period: A Pedoclimatic Discrimination

The volatile profile of an olive oil is a crucial attribute indicating its sensory quality. Hence, to elucidate the impact of geographical origin (including edaphoclimatic conditions) and the crop season on the volatile composition of monovarietal Moroccan olive oil “Picholine Marocaine”, over a two-year harvest period (2018/19 and 2019/20), thirty-eight olive oil samples were obtained from nineteen Moroccan areas well-known by the abundance of olive tree cultivation. By using SPME/GC-FID-MS, 229 and 215 volatile compounds were characterized in olive oils produced during the 2018/19 and 2019/20 crop years, respectively. The identified compounds belong to nine volatile groups: terpenes, hydrocarbons, furans, esters, alcohols, acids, ketones, aldehydes, and nitrogen compounds. The one-way ANOVA and interactive heatmap revealed significant differences in the volatiles proportion in oils from different geographical origins. Our results imply that environmental (edaphoclimatic) conditions considerably influence the volatile compounds’ biosynthesis, e.g., when soil granulometry decreases (from sand to silt), alcohols become esters due to the activity of alcohol dehydrogenase (ADH) and alcohol acetyl transferase (AAT) enzymes. Moreover, our findings exhibit a significant influence of the crop season on the volatile composition of Moroccan olive oils.

Metabolic flux analysis of the effect of carbon dioxide top pressure on acetyl coenzyme A and ester production by Saccharomyces cerevisiae

ABSTRACT Carbon dioxide can affect the growth of beer yeast, the production of acetyl-CoA and the synthesis of esters. Through the analysis of metabolic flux, the influence of CO2 top pressure application on the flow direction of carbon metabolism in beer fermentation and the key sites of its action, especially the flow direction of the substrate acetyl-CoA, were studied. It was found that the CO2 top pressure inhibited the formation of acetyl-CoA and higher alcohols but barely affected the activity of alcohol acetyltransferase (AATase). Carbon dioxide top pressure application increased the permeation of acetic acid to extracellular environments, decreased the flux of carbon metabolism toward acetyl-CoA production, and, finally, reduced the synthesis of total esters in beer. Carbon dioxide top pressure application enhanced the pyruvate carboxylation of oxaloacetic acid nodes, while acetyl-CoA, involved in biosynthesis, including that of fatty acids, was inhibited.

Expansion of PmBEAT genes in the Prunus mume genome induces characteristic floral scent production

Prunus mume is the only plant in the genus Prunus of the Rosaceae family with a characteristic floral scent, and the main component of this scent is benzyl acetate. By contrast, benzyl acetate is not synthesized in Prunus persica flowers. Here, we searched for benzyl alcohol acetyltransferase (BEAT) genes based on genomic data from P. mume and P. persica and found 44 unique PmBEATs in P. mume. These genes, which were mainly detected in clusters on chromosomes, originated from gene duplication events during the species evolution of P. mume, and retroduplication and tandem duplication were the two dominant duplication patterns. The genes PmBEAT34, PmBEAT36 and PmBEAT37, which were generated by tandem duplication, were highly expressed in flowers, and their highest levels were detected during the blooming stage. In vitro, PmBEAT34, PmBEAT3, and PmBEAT37 all had benzyl alcohol acetyltransferase activity that was localized in the cytoplasm. Overexpression of the PmBEAT36 or PmBEAT37 genes increased benzyl acetate production in the petal protoplasts of P. mume, and interference in the expression of these genes slightly decreased the benzyl acetate content. In addition, light and temperature regulated the expression of the PmBEAT34, PmBEAT36 and PmBEAT37 genes. According to these results, we hypothesize that the expansion of the PmBEAT genes in the genome induce the characteristic floral scent of P. mume.

Acetate-ester hydrolase activity for screening of the variation in acetate ester yield of Cyberlindnera fabianii, Pichia kudriavzevii and Saccharomyces cerevisiae

Esters constitute an important class of aroma compounds and contribute significantly to the aroma of yeast-fermented beverages. Ester formation is well studied in Saccharomyces cerevisiae, while production of aroma compounds by non-conventional yeasts has received little attention. The selection of such strains for co-culturing with S. cerevisiae offers opportunities for product innovations. Therefore, we performed a comparative analysis of the diversity in ester production by Cyberlindnera fabianii 65 (Cf65), Pichia kudriavzevii 129 (Pk129) and S. cerevisiae 131 (Sc131). For all three species distinct aroma profiles were identified, with Cf65 producing the highest amount of acetate esters. Since esters are formed from alcohols and acyl-CoA or acetyl-CoA, we analysed in vitro alcohol dehydrogenase and alcohol acetyl transferase activities in those three yeasts and found no correlation with ester formation. In contrast, a clear inverse correlation between the acetate-ester hydrolase activity and acetate ester yield was found for the three yeast species. Our study indicates that acetate-ester hydrolase activity plays a key role in determination of the final amount of acetate esters in fermentation broths.

PGK1 Promoter Library for the Regulation of Acetate Ester Production in Saccharomyces cerevisiae during Chinese Baijiu Fermentation.

Appropriate concentrations and proportion of acetate esters and higher alcohols improve the quality of Chinese Baijiu. To regulate the concentrations of acetate esters in Chinese Baijiu, we constructed a PGK1 promoter library through error-prone PCR. Then, we used an enhanced green fluorescent protein as a reporter to characterize the activities of PGK1p mutants. The PGK1p library contained 28 PGK1p mutants and spanned an activity that ranged between 0.1% and 141% of wild-type PGK1p. Seven PGK1p mutants were characterized by an additional reporter β-galactosidase and then used for the overexpression of ATF1 with BAT2 deletion in Saccharomyces cerevisiae a45. The production of ethyl acetate in strains A8, A17, A18, A27, A22, A25, A28, and AWT were 1.66-, 3.09-, 10.59-, 13.07-, 15.99-, 22.67-, 24.06-, and 27.22-fold higher than that of the parental strain. The results on alcohol acetyltransferase (AATase) activity showed that the PGK1p library precisely controlled ATF1 expression and regulated the acetate esters production.

Concealed ester formation and amino acid metabolism to volatile compounds in table grape (Vitis vinifera L.) berries

Volatile esters contribute to the aroma and flavor of many fruits but are normally absent in grape berries (Vitis vinifera L.). To examine the biosynthetic potential of grape berries to form volatile esters, berry sections were incubated with exogenous L-Phe, L-Leu or L-Met. In general, amino-acid incubation caused the accumulation of the respective aldehydes and alcohols. Moreover, L-Leu incubation resulted in the accumulation of 3-methylbutyl acetate and L-Phe incubation resulted in the accumulation 2-phenylethyl acetate in ‘Muscat Hamburg’ but not in the other grape accessions. Exogenous L-Met administration did not result in volatile esters accumulation but the accumulation of sulfur volatile compounds such as methional and dimethyl disulfide was prominent. Berry-derived cell-free extracts displayed differential alcohol acetyltransferase activities and supported the formation of 3-methylbutyl acetate and benzyl acetate. 2-Phenylethyl acetate was produced only in ‘Muscat Hamburg’ cell-free extracts. VvAAT2, a newly characterized gene, was preferentially expressed in ‘Muscat Hamburg’ berries and functionally expressed in E. coli. VvAAT2 possesses alcohol acetyltransferase activity utilizing benzyl alcohol, 2-phenylethanol, hexanol or 3-methylbutanol as substrates. Our study demonstrates that grape berries have a concealed potential to accumulate volatile esters and this process is limited by substrate availability.

Ethyl acetate production by the elusive alcohol acetyltransferase from yeast

Ethyl acetate is an industrially relevant ester that is currently produced exclusively through unsustainable processes. Many yeasts are able to produce ethyl acetate, but the main responsible enzyme has remained elusive, hampering the engineering of novel production strains. Here we describe the discovery of a new enzyme (Eat1) from the yeast Wickerhamomyces anomalus that resulted in high ethyl acetate production when expressed in Saccharomyces cerevisiae and Escherichia coli. Purified Eat1 showed alcohol acetyltransferase activity with ethanol and acetyl-CoA. Homologs of eat1 are responsible for most ethyl acetate synthesis in known ethyl acetate-producing yeasts, including S. cerevisiae, and are only distantly related to known alcohol acetyltransferases. Eat1 is therefore proposed to compose a novel alcohol acetyltransferase family within the α/β hydrolase superfamily. The discovery of this novel enzyme family is a crucial step towards the development of biobased ethyl acetate production and will also help in selecting improved S. cerevisiae brewing strains.

Differences in Enzymatic Properties of the Saccharomyces kudriavzevii and Saccharomyces uvarum Alcohol Acetyltransferases and Their Impact on Aroma-Active Compounds Production.

Higher alcohols and acetate esters belong to the most important yeast secondary metabolites that significantly contribute to the overall flavor and aroma profile of fermented products. In Saccharomyces cerevisiae, esterification of higher alcohols is catalyzed mainly by the alcohol acetyltransferases encoded by genes ATF1 and ATF2. Previous investigation has shown other Saccharomyces species, e.g., S. kudriavzevii and S. uvarum, to vary in aroma-active higher alcohols and acetate esters formation when compared to S. cerevisiae. Here, we aimed to analyze the enzymes encoded by the ATF1 and ATF2 genes from S. kudriavzevii (SkATF1, SkATF2) and S. uvarum (SuATF1, SuATF2). The heterologous expression of the individual ATF1 and ATF2 genes in a host S. cerevisiae resulted in the enhanced production of several higher alcohols and acetate esters. Particularly, an increase of 2-phenylethyl acetate production by the strains that harbored ATF1 and ATF2 genes from S. kudriavzevii and S. uvarum was observed. When grown with individual amino acids as the nitrogen source, the strain that harbored SkATF1 showed particularly high 2-phenylethyl acetate production and the strains with introduced SkATF2 or SuATF2 revealed increased production of isobutyl acetate, isoamyl acetate, and 2-phenylethyl acetate compared to the reference strains with endogenous ATF genes. The alcohol acetyltransferase activities of the individual Atf1 and Atf2 enzymes measured in the cell extracts of the S. cerevisiae atf1 atf2 iah1 triple-null strain were detected for all the measured substrates. This indicated that S. kudriavzevii and S. uvarum Atf enzymes had broad range substrate specificity as S. cerevisiae Atf enzymes. Individual Atf1 enzymes exhibited markedly different kinetic properties since SkAtf1p showed c. twofold higher and SuAtf1p c. threefold higher Km for isoamyl alcohol than ScAtf1p. Together these results indicated that the differences found among the three Saccharomyces species during the aroma-active acetate ester formation may be due, to some extent, to the distinct properties of Atf enzymes.

A novel method to quantify the activity of alcohol acetyltransferase Using a SnO2-based sensor of electronic nose

Alcohol acetyltransferase (AATFase) extensively catalyzes the reactions of alcohols to acetic esters in microorganisms and plants. In this work, a novel method has been proposed to quantify the activity of AATFase using a SnO2-based sensor of electronic nose, which was determined on the basis of its higher sensitivity to the reducing alcohol than the oxidizing ester. The maximum value of the first-derivative of the signals from the SnO2-based sensor was therein found to be an eigenvalue of isoamyl alcohol concentration. Quadratic polynomial regression perfectly fitted the correlation between the eigenvalue and the isoamyl alcohol concentration. The method was used to determine the AATFase activity in this type of reaction by calculating the conversion rate of isoamyl alcohol. The proposed method has been successfully applied to determine the AATFase activity of a cider yeast strain. Compared with GC–MS, the method shows promises with ideal recovery and low cost.

Influence of carbon and nitrogen source on production of volatile fragrance and flavour metabolites by the yeast Kluyveromyces marxianus.

The yeast Kluyveromyces marxianus produces a range of volatile molecules with applications as fragrances or flavours. The purpose of this study was to establish how nutritional conditions influence the production of these metabolites. Four strains were grown on synthetic media, using a variety of carbon and nitrogen sources and volatile metabolites analysed using gas chromatography-mass spectrometry (GC-MS). The nitrogen source had pronounced effects on metabolite production: levels of the fusel alcohols 2-phenylethanol and isoamyl alcohol were highest when yeast extract was the nitrogen source, and ammonium had a strong repressing effect on production of 2-phenylethyl acetate. In contrast, the nitrogen source did not affect production of isoamyl acetate or ethyl acetate, indicating that more than one alcohol acetyl transferase activity is present in K. marxianus. Production of all acetate esters was low when cells were growing on lactose (as opposed to glucose or fructose), with a lower intracellular pool of acetyl CoA being one explanation for this observation. Bioinformatic and phylogenetic analysis of the known yeast alcohol acetyl transferases ATF1 and ATF2 suggests that the ancestral protein Atf2p may not be involved in synthesis of volatile acetate esters in K. marxianus, and raises interesting questions as to what other genes encode this activity in non-Saccharomyces yeasts. Identification of all the genes involved in ester synthesis will be important for development of the K. marxianus platform for flavour and fragrance production.

Residual effects of low oxygen storage of mature green fruit on ripening processes and ester biosynthesis during ripening in bananas

Mature green banana (Musa sapientum L. cv. Cavendish) fruit were stored in 0.5%, 2%, or 21% O2 for 7 days at 20°C before ripening was initiated by ethylene. Residual effects of low O2 storage in mature green fruit on ripening and ester biosynthesis in fruit were investigated during ripening for up to 6 d at 20°C. Concentrations of ethanol in mature green fruit did not change during storage in both 21% and 2% O2 atmospheres, but increased in fruit stored in 0.5% O2. The activities of alcohol dehydrogenase (ADH) in 2% and 21% O2 atmospheres remained very low throughout the storage period, but significantly increased with 0.5% O2. After transferring fruit to regular air and trigging ripening with ethylene, yellowing of peel, fruit softening and hydrolysis of starch in fruit stored in low O2 atmospheres were slower than in the control. Fruit stored in low O2 also showed a delayed onset of the climacteric peak. The activities of ADH were lower in the low O2 stored fruit than in the control fruit. Productions of ethyl acetate, isoamyl acetate, and isobutyl acetate were remarkably suppressed by low O2 storage. Alcohol acetyltransferase activity increased gradually with storage time in all treatments, being significantly lower in fruit with low O2 pretreatments. The results indicate that low O2 plus room temperature storage can extend storage life of bananas with the sacrifice of a low production of ester volatiles.

Antisense suppression of alcohol acetyltransferase gene in ripening melon fruit alters volatile composition

Volatile esters are characteristic aroma components of melon (Cucumis melo L.) fruit. Alcohol acetyltransferase (AAT) (EC2.3.1.84) catalyzes the reactions of aliphatic and aromatic alcohols to their esters in the presence of acyl CoA in fruit and flowers. In this study, melon plants were transformed with a gene construct containing a melon AAT cDNA (AAT2) in an antisense orientation, driven by the constructive cauliflower mosaic virus 35S promoter, by ovary-injection transformation method. The transformed plants displayed a range of decreased AAT activities in the ripening fruit. The relative content of volatile esters was reduced, and the average relative content in the transgenic fruit was 78% of the wild-type fruit. However, the relative contents of aldehydes and alcohols increased; the average relative contents were 3.2 times and 2.4 times of those in the wild-type fruit, respectively. Apart from the difference in the volatile compounds, no other phenotypic alterations were noted, including other quality indices, such as the sugar, vitamin C and carotenoid contents. Our study indicates that AAT plays the key role in volatile ester formation in melon fruit.

Characterization and Isolation of Mutants Producing Increased Amounts of Isoamyl Acetate Derived from Hygromycin B-Resistant Sake Yeast

Hygromycin B is an aminoglycoside antibiotic that inhibits protein synthesis in prokaryotes and eukaryotes. Twenty-four hygromycin B-resistants mutants were isolated from sake yeast, and were divided into three different degrees of strength according to hygromycin B resistance. Three of four hygromycin B strongly resistant mutants produced increased amounts of isoamyl acetate in sake brewing test, although isoamyl alcohol levels remained unchanged. Many hygromycin B-resistants mutants showed higher E/A ratios than K-701 in culture with koji extract medium. Strain HMR-18 produced the largest amount of isoamyl acetate, and its alcohol acetyltransferase (AATFase) activity was 1.3-fold that of K-701. DNA microarray analysis showed that many genes overexpressed in HMR-18 were involved in stress responses (heat shock, low pH, and so on) but HMR-18 showed thermo- and acid-sensitivity. It was strongly resistant to hygromycin B and another aminoglycoside antibiotic, G418.

Effects of Deep Sea Water (DSW) Nigari on the Alcohol Production of Yeasts Fermented in Acidic Roselle Extract

The effects of deep sea water (DSW) nigari refined from DSW pumped from a depth of 400 m off Hualien, Taiwan, on the brewer's yeasts used in making a beverage based on roselle extract (pH 2.57) were investigated. Experimental results showed that the addition of DSW nigari (300 mg/L) produced a higher alcohol yield with yeast strains B1 (4.13% [vol/vol]), B6 (4.97% [vol/vol]), and Saccharomyces bayanus BCRC 21673 (3.00% [vol/vol]), in marked contrast to not adding DSW nigari to the three yeast strains (2.23, 1.77, and 0.77% [vol/vol] for B1, B6, and BCRC 21673, respectively). The results also indicated that the increase in alcohol yield was dose dependent when DSW nigari was used at 0, 150, 300, and 450 mg/L with strain B6. Although the addition of DSW nigari resulted in higher levels of viable cells than those in the control samples, they did not significantly differ from each other. A concentration of four individual metal chlorides (CaCl2, KCl, MgCl2, and NaCl) or a mixture of these metals equivalent to 300 mg of DSW nigari per L was added to investigate which major components of DSW nigari affect strain B6 with regard to growth and the production of alcohol and flavor compounds. The results revealed that Mg and Ca did not affect yeast growth, whereas K, Na, and the metal mixture did, although they were still less effective than DSW nigari. Furthermore, DSW nigari produced the highest yield of alcohol, followed by the metal mixture. No significant differences were observed between the individual metals and the control group. Addition of DSW nigari increased the amounts of ethyl acetate, isoamyl alcohol, and isobutanol produced; however, this phenomenon was not directly due to stimulation from alcohol acetyltransferase activity but was caused mainly by an increase in yeast growth.

Enantioselective monoterpene alcohol acetylation in Origanum, Mentha and Salvia species

Selected plants within the Origanum, Mentha and Salvia genera, that contain significant amounts of chiral volatile alcohols and their related acetates, exhibit remarkable enantioselectivity of alcohol acetyl transferase (AAT) activity and particularly can discriminate between linalool enantiomers. Origanum dayi AAT produced almost enantiomerically pure ( R)-linalyl acetate by enzymatic acetylation of racemic linalool, whereas the closely related O. majorana AAT produced a mixture of ( R)- and ( S)-linalyl acetate with a ratio of 6:4. V max of O. dayi acetylation activity was 30-fold higher for ( R)-linalool, whereas in O. majorana no such differences were found.

Effect of short‐term anaerobic conditions on the production of volatiles, activity of alcohol acetyltransferase and other quality traits of ripened bananas. Sumithra K Wendakoon. Yoshinori Ueda. Yoshihiro Imahori. Megumi Ishimaru. Journal of the Science of Food and Agriculture, 2006; 86: 1475–1480

AbstractThe original article to which this Erratum refers was published in Journal of the Science of Food and Agriculture, 2006; 86: 1475–1480.

The effect of increased yeast alcohol acetyltransferase and esterase activity on the flavour profiles of wine and distillates

The fruity odours of wine are largely derived from the synthesis of esters and higher alcohols during yeast fermentation. The ATF1- and ATF2-encoded alcohol acetyltransferases of S. cerevisiae are responsible for the synthesis of ethyl acetate and isoamyl acetate esters, while the EHT1-encoded ethanol hexanoyl transferase is responsible for synthesizing ethyl caproate. However, esters such as these might be degraded by the IAH1-encoded esterase. The objectives of this study were: (a) to overexpress the genes encoding ester-synthesizing and ester-degrading enzymes in wine yeast; (b) to prepare Colombard table wines and base wines for distillation using these modified strains; and (c) to analyse and compare the ester concentrations and aroma profiles of these wines and distillates. The overexpression of ATF1 significantly increased the concentrations of ethyl acetate, isoamyl acetate, 2-phenylethyl acetate and ethyl caproate, while the overexpression of ATF2 affected the concentrations of ethyl acetate and isoamyl acetate to a lesser degree. The overexpression of IAH1 resulted in a significant decrease in ethyl acetate, isoamyl acetate, hexyl acetate and 2-phenylethyl acetate. The overexpression of EHT1 resulted in a marked increase in ethyl caproate, ethyl caprylate and ethyl caprate. The flavour profile of the wines and distillates prepared using the modified strains were also significantly altered as indicated by formal sensory analysis. This study offers prospects for the development of wine yeast starter strains with optimized ester-producing capability that could assist winemakers in their effort to consistently produce wine and distillates such as brandy to definable flavour specifications and styles.

Cold Storage Effects on Pawpaw Fruit Ripening and Quality

Pawpaw [Asimina triloba (L.) Dunal] has significant potential as a new fruit crop. During ripening, loss of firmness is extremely rapid, and this trait may be the biggest obstacle to the development of a broader market. Cold storage of pawpaw fruit seems limited to 4 weeks at 4 °C, though fruit softening merely slows during storage. A study of several cultivars with commercial appeal has shown that none have superior cold storage life. Extending the cold storage beyond 4 weeks resulted in increasing loss of fruit firmness, poor poststorage ripening, and development of quality traits, and many fruit exhibiting flesh and peel discoloration. Cold storage duration affected fruit volatile production. By 4 weeks of cold storage, ethyl hexanoate and ethyl octanoate were the major volatiles produced, replacing methyl hexanoate, which was the major volatile produced by ripe fruit after harvest. By 8 weeks of cold storage, volatile ester production was generally low and ethyl hexanoate became the only major volatile. This loss of volatile production was accompanied by a decrease in alcohol acetyl transferase activity. Also, during cold storage, there was an increase in total phenolic content, lipid peroxidation products, and polyphenol oxidase activity. These changes may contribute to the black discoloration that developed in fruit cold-stored for 8 weeks or more. It is apparent that cold storage alone may not be sufficient to extend the storage life of most, if not all, current pawpaw cultivars beyond 4 weeks.

Effect of short‐term anaerobic conditions on the production of volatiles, activity of alcohol acetyltransferase and other quality traits of ripened bananas

AbstractYellow, ripened (yellow with green tips) bananas were treated with nitrogen gas for 12, 24 and 48 h at 20 °C to study the effects of anaerobic conditions on the fruit quality, including the production of volatile compounds and activity of alcohol acetyltransferase (AAT) during the post‐harvest period. Significantly higher concentrations of acetaldehyde and ethanol were found in the treated fruit than in the untreated bananas after removal from the conditions. The 48‐h treatment showed higher levels of ethanol and acetaldehyde after storage. Immediately after releasing the fruit from the anaerobic atmosphere, the amounts of isobutyl acetate and isoamyl acetate decreased. The production of ethyl acetate increased markedly in each treatment unit 1 day after removing the fruit. Regardless of recovery from the production of isobutyl acetate and isoamyl acetate 1 day after treatment, the banana‐like aroma was denatured because of the production of high levels of ethyl acetate and ethanol. The increase in AAT was slightly lower in nitrogen‐treated fruit than untreated fruit. However, the activity in treated fruit was sufficient for ester production in bananas. The endogenous alcohol levels were increased during nitrogen gas treatment in the fruit due to the inhibition of ester production. Colour development of the bananas was greatly inhibited by the treatments, while the sugar content did not show any differences between the treated and untreated fruit. The results suggest that, even if bananas are removed from the short‐term anaerobic conditions, a loss in quality, except sweetness, easily occurs afterwards, especially the characteristic aroma of the fruit during the post‐harvest period. Copyright © 2006 Society of Chemical Industry