Acetate Esters Research Articles

Bridging the Gap: linking Torulaspora delbrueckii Genotypes to Fermentation Phenotypes and Wine Aroma.

Climate change and consumer preferences are driving innovation in winemaking, with a growing interest in non-Saccharomyces species. Among these, Torulaspora delbrueckii (Td) has gained recognition for its ability to reduce volatile acidity and enhance aromatic complexity in wine. However, knowledge regarding its phenotypic and genomic diversity impacting alcoholic fermentation remains limited. Aiming to elucidate the metabolic differences between Td and Saccharomyces cerevisiae (Sc) and the Td intraspecies diversity, we conducted a comprehensive metabolic characterization of fifteen Td strains. This analysis delved beyond standard fermentation parameters (kinetics and major metabolites production) to explore non-conventional aromas and establish genotype-phenotype links. Our findings confirmed that most Td strains produce less acetic acid and more succinate and glycerol than Sc. The overall aromatic profiles of Td strains differed from Sc, exhibiting higher levels of monoterpenes and higher alcohols, while producing less acetate esters, fatty acids, their corresponding ethyl esters, and lactones. Moreover, we identified the absence of genes responsible for specific aroma profiles, such as decreased ethyl esters production, as well as the absence of cell wall genes, which might negatively affect Td performance when compared to Sc. This work highlights the significant diversity within Td and underscores potential links between its genotype and phenotype.

Systematic study on the hydrogen abstraction reactions from oxygenated compounds by H and HO2

AbstractTo extend the rule‐based approach for hydrogen abstraction reactions from oxygenated compounds, a systematic investigation was performed to examine the reactivity of gas‐phase hydrogen abstraction reactions from alkyl groups (methyl and ethyl groups) bound to oxygen atoms in five types of oxygenated compounds (alcohols, ethers, formate esters, acetate esters, and carbonate esters) by H atoms and HO2 radicals comprehensively considering rotational conformers. Quantum chemical calculations were conducted at the CBS‐QB3 level for stationary points. Rate constants were determined employing conventional transition state theory (TST). For hydrogen abstraction reactions by H, the rotational conformer distribution partition function was employed to approximate partition functions, owing to the similarity in vibrational energy‐level structures among conformers. In hydrogen abstraction reactions by HO2, the vibrational structures of transition‐state (TS) conformers varied significantly due to the hydrogen bonding, leading to an inappropriate evaluation of rate constants when using the lowest‐energy conformer as a representative. Therefore, the rate constants were calculated by the multi‐structural TST. It was revealed that the differences in functional groups containing O atoms mainly affect the bond dissociation energies of the C–H bonds and the activation energies of hydrogen abstraction reactions only when the C atoms are adjacent to the O atoms. Additionally, it was found that hydrogen bonds formed in the TSs show minor effect on rate parameters for the overall rate constants, apart from the reduction of the pre‐exponential factors for the H‐abstraction reactions from the methylene position of ethyl groups. The comparison with the rate constants from previous studies showed reasonable results, indicating that the rate constants in this study, which thoroughly consider rotational conformers, can be the current best estimates.

Study on Evaluation of Fruit Aroma of Plum Variety Resources Based on Headspace Solid-Phase Microextraction Combined with Gas Chromatography-Mass Spectrometry.

To explore the characteristics of and variations in aroma components across different plum varieties and maturity stages, this study employed headspace solid-phase microextraction combined with gas chromatography-mass spectrometry (HS-SPME-GC-MS). This method was used to systematically analyze the aroma components of 12 early-maturing, 15 medium-maturing, and 11 late-maturing plum varieties. The variations in volatile components among these three germplasm types were then compared using multivariate statistical methods. The examination revealed that 138 aromatic components were meticulously identified and quantified, such as 26 aldehydes, 63 esters, 13 ketones, 30 alcohols, and six other compounds. Thirteen main aroma compounds including acetic acid hexyl ester, (Z)-3-hexen-1-ol acetate, hexanal, 1-hexanol, 3-hexenal, butanoic acid butyl ester, (E)-2-hexen-1-ol, hexanoic acid butyl ester, propanoic acid butyl ester, (E)-2-hexenal, L-.alpha.-terpineol, (Z)-2-hexen-1-ol acetate, and 1-butanol were considered dominant. The orthogonal partial least squares discriminant analysis (OPLS-DA) combined with variable importance projection (VIP) results showed that 24 differential aroma compounds were screened out from 38 varieties of plum fruits based on their differences in aroma components, which can be used to distinguish plum fruits at different ripening times. Twenty-four aroma-contributing compounds were identified based on their odor activity values (OAVs). Among these, 14 key aroma components with OAVs ≥ 10 were highlighted. In summary, the aroma compounds of early- and late-maturing plum germplasm exhibited rich diversity, with significant differences in aroma components between plums of varying maturity and germplasm. These differences can serve as indicators for identifying different plum germplasm.

Rapid in situ carbon-13 hyperpolarization and imaging of acetate and pyruvate esters without external polarizer

Hyperpolarized 13C MRI visualizes real-time metabolic processes in vivo. In this study, we achieved high 13C polarization in situ in the bore of an MRI system for precursor molecules of most widely employed hyperpolarized agents: [1-13C]acetate and [1-13C]pyruvate ethyl esters in their perdeuterated forms, enhancing hyperpolarization lifetimes, hyperpolarized to P13C ≈ 28% at 80 mM concentration and P13C ≈ 19% at 10 mM concentration, respectively. Using vinyl esters as unsaturated Parahydrogen-Induced Polarization via Side-Arm Hydrogenation (PHIP-SAH) precursors and our novel polarization setup, we achieved these hyperpolarization levels by fast side-arm hydrogenation in acetone-d6 at elevated temperatures (up to 90°C) and hydrogenation pressures (up to 32 bar). We optimized the hyperpolarization process, reducing it to under 10 s, and employed advanced pulse sequences to enhance the polarization transfer efficiency. The hyperpolarization system has a small footprint, allowing it to be positioned in the same magnet, where 13C MRI is performed. We exemplified the utility of the design with sub-second in situ 13C MRI of ethyl [1-13C]pyruvate-d6. However, challenges remain in side-arm cleavage and purification in the MRI system to extract highly polarized aqueous agent solutions. Our results showcase efficient and rapid 13C hyperpolarization of these metabolite precursors in an MRI system with minimal additional hardware, promising to enhance future throughput and access to hyperpolarized 13C MRI.

The Role of Keeving in Modulating Fermentation and the Flavour Profiles of Apple Brandy

Keeving is the removal of nutrients from apple musts due to their binding to pectin, resulting in a slower fermentation and spontaneous arrest. The aim of this study was to determine the effect of keeving on the chemical composition of fermented apple must and on the volatile profile and sensory analysis of apple brandies. We compared the application of keeving during spontaneous fermentation with fermentation carried out by Saccharomyces cerevisiae (SafSpirit HG-1). We evaluated the impact of adding different doses of calcium chloride on various parameters of fermented musts and distillates. Calcium chloride had a greater effect on the ethanol concentration, total extract, and fermentation efficiency than on the type of fermentation used. However, a different phenomenon was observed with respect to the volatiles. The concentration of most of the higher alcohols, acetaldehyde, dodecanal, and geranylaceton, decreased after spontaneous fermentation and increased during the fermentation carried out with Saccharomyces cerevisiae SafSpirit HG-1. In general, the application of keeving contributed to a decrease in the concentration of ethyl and methyl esters, but caused an increase in the concentration of all acetate esters and terpenoids. When the amount of nutrients in the environment is limited and starvation occurs, microorganisms use the available nutrients for basic metabolic processes that allow them to survive and limit the formation of side metabolites such as volatiles. However, most of the samples fermented after the faecal depletion achieved high scores for the floral, fruity, and “overall note” parameters in the sensory analysis. This means that this method, carried out with a properly selected yeast strain, could be feasible for the distilling industry.

Characterization of Changes in Ripening Process of Volatile Apple Compounds Based on HS-SPME-GC-MS Analysis

The aim of this study was to identify the aromatic compounds present in different apple varieties and to gain insights into the changes in the aromatic compounds during ripening. Three apple varieties (“Red Astrachan”, “Ning Qiu”, and “Golden Delicious”) at different stages of ripening were selected for this study; their peel and pulp were analyzed via headspace solid-phase microextraction (HS-SPME)–gas chromatography–mass spectrometry (HS-SPME-GC-MS), and 30 volatile compounds were identified. The samples’ differences were analyzed using heat map cluster analysis, principal component analysis (PCA), and an independent samples t-test. The results showed that the content of aromatic compounds in the peel was higher than that in the pulp. The relative content of esters in the aromatic compounds of the three apple varieties followed the order of pulp > peel and “Ning Qiu” > “Golden Delicious” > “Red Astrachan”. This suggests that “Ning Qiu” combines the advantages of its parents in terms of its aroma content. The highest concentrations of aroma compounds in “Red Astrachan” and “Ning Qiu” accumulate before the ripening stage, and care should be taken to choose an appropriate harvesting time according to the different needs during production. The main compounds of “Red Astrachan” are aldehydes and C8 esters, while those of “Ning Qiu” and “Golden Delicious” are alkenes and esters. After analyzing the relative odor activity values (rOAVs) of key volatile compounds and their aroma descriptors during the harvest period, acetic acid pentyl ester, butanoic acid hexyl ester, hexanoic acid hexyl ester, and 2-ethyl-1-Hexanol were found to contribute the most to the overall flavor of the peel and pulp. “Ning Qiu” combines the parental advantages of the concentrated peel of “Red Astrachan” and the astringent pulp of “Golden Delicious”, with compounds in its composition that give a pleasing aroma. Mature “Ning Qiu” fruits have a more intense aroma and fruity flavor. The development of flavor-specific varieties has provided the theoretical basis for future research in molecular hybridization, molecular-assisted breeding, and the molecular biology of apple flavor synthesis and metabolism.

Enhancing the fermentation rate and aroma compounds of kiwi wine via the addition of nitrogen during the stationary phase

Nitrogen addition is a viable strategy for preventing sluggish or stuck fermentation and enhancing aroma profiles in the winemaking industry. Herein, nitrogen, in the form of amino acids mixture or diammonium hydrogen phosphate (DAP), was added at different fermentation periods to investigate its effects on the fermentation characteristics, non-volatile compounds and aroma profiles of kiwi wines. Both the amino acids mixture and DAP shortened fermentation time by 1–2 days and balanced organic acid proportion by reducing malic acid content and increasing citric acid content. Adding DAP significantly decreased higher alcohols content, whereas higher alcohols content was remarkably increased with amino acids mixture supplementation. Furthermore, nitrogen addition during the stationary phase was most conducive to synthesizing other beneficial aromas, especially acetate esters and medium-chain fatty acid ethyl esters (MCFAEEs), and improving the perception of tropical fruity and cream aromas in kiwi wines. Partial least-squares regression (PLSR) results showed there was a tight correlation between amino acids and volatile compounds. This study provides an insight into the development of high-quality kiwi wine and should serve as the theoretical foundation to support the application of nitrogen addition in the wine industry.

Design, Synthesis, and Antimicrobial Evaluation of Novel Schiff Bases Derived from Thymol

Schiff bases, a significant class of organic compounds, are synthesized through the reaction of a primary amine with an aldehyde or ketone under specific conditions. In this study, we report the synthesis of novel Schiff bases derived from thymol (2-Isopropyl-5-methylphenol). The synthesis began with the esterification of thymol to produce (2-Isopropyl-5-methylphenoxy)-acetic acid ethyl ester (1), followed by hydrazination to yield (2-Isopropyl-5-methylphenoxy)-acetic acid hydrazide (2). The hydrazide (2) was then treated with various aromatic aldehydes to synthesize a series of Schiff bases (3A-J). The chemical structures of these compounds were elucidated using infrared (IR) spectroscopy, proton nuclear magnetic resonance (1H-NMR), and mass spectrometry. The synthesized Schiff bases were evaluated for their antibacterial and antifungal activities using standard screening methods. The results indicated that all synthesized compounds exhibited varying degrees of antimicrobial activity. Among the series, compound 3C showed the highest antibacterial and antifungal potency, suggesting it could be a promising candidate for further development. This study highlights the potential of Schiff bases as bioactive compounds with broad-spectrum antimicrobial properties. The structure-activity relationship (SAR) suggests that the presence of specific substituents on the aromatic aldehyde moiety could enhance biological activity. Further investigation into the mechanisms of action and optimization of these Schiff bases could contribute to the development of novel antimicrobial agents

Effect of Grist Particle Size Distribution and Wort Turbidity on Ester Composition of Malt Whisky New Make Spirit

The parameters used during milling of malt have considerable implications for the turbidity of wort collected during lautering, and wort turbidity has previously been demonstrated to impact ester development by yeast (during fermentation of brewer’s wort). The present research assesses the impact of milling regime and wort turbidity on congener development during production of malt whisky new make spirit. New make spirit was produced from pot still malt prepared with a disc mill under differing milling regimes (mill gap: 0.5–1.5 mm; laboratory scale). Use of increasingly coarse milling conditions positively correlated with wort turbidity and reduced concentration of acetate and ethyl esters in new-make spirit. Similar to previous research within the brewing industry, turbid wort was generally associated with reduced ester content, particularly in samples produced under the coarsest milling conditions (mill gap: 1.5 mm). Turbidity of new-make spirit (diluted to 30% ABV) was reduced in samples produced from wort of elevated turbidity, likely due to reduced content of ethyl esters and other haze-active congeners.

Xylan acetate ester ameliorates ulcerative colitis through intestinal barrier repair and inflammation inhibition via regulation of macrophage M1 polarization

Ulcerative colitis (UC) is a chronic inflammatory disease resulting from abnormal immune response to gut microflora translocating through damaged intestinal barrier. Xylan acetate ester (XylA) can increase colon short-chain fatty acids (SCFAs) levels and alleviate kidney disease by inhibiting inflammation through the G protein-coupled receptor pathway. Here, we synthesized and purified XylA, and then the effects and mechanisms of XylA on dextran sodium sulfate-induced UC in mice were investigated. The results showed that in mice, similar to the positive drug 5-aminosalicylic acid, oral administration of XylA significantly alleviated all UC symptoms, including weight loss, diarrhea, and hematochezia. Further mechanism studies revealed that XylA could repair the damaged colon structure and intestinal barrier function by increasing the expression of tight junction protein zonula occludens 1 and occludin, thus reducing LPS penetration. Moreover, XylA could also restrain intestinal inflammation via inhibiting LPS-TLR4 pathway, downregulating M1 macrophage polarization, and reducing proinflammatory cytokines expression, and in vitro cell experiments showed that these effects may be mediated by XylA derived SCFAs, particularly acetates, propionates and butyrates. All these results suggested that XylA may be a potential improving agent for UC treatment, and natural polysaccharides may represent a novel avenue for drug development of UC.

Antimicrobial metabolites from Probiotics, Pleurotus ostreatus and their co-cultures against foodborne pathogens isolated from ready-to-eat foods

BackgroundThe incidence of foodborne pathogens in ready- to-eat (RTE) can be attributed to various foodborne diseases. Most of the isolated microorganisms from RTE foods are resistant to common antibiotics and thus, resulted to treatment failure when commercially available antibiotics are administered. However, the secondary metabolites secreted by microorganisms can serve as alternative therapy that are reliable and safe. Secondary metabolites obtained from mono- and co-culture microorganisms can inhibit the growth of antibiotic-resistant microorganisms. Bioactive compounds in the secreted metabolites can be identified and utilized as sources of new antibiotics. In this study, antimicrobial activity of secondary metabolites from Lactobacillus fermentum, Saccharomyces cerevisiae, Pleurotus ostreatus, and their co-cultures were tested against foodborne pathogens isolated from RTE foods using agar well diffusion. The bioactive compounds in the metabolites were identified using gas chromatography-mass spectrometry.ResultsFrom a total of 100 RTE foods examined, Salmonella enterica, Shigella dysenteriae, Escherichia coli, Klebsiella pneumoniae (subsp ozaenae), Pseudomonas fluorescens, Clostridium perfringes, Bacillus cereus, Listeria monocytogens, and Staphylococcus aureus, Penicillium chrysogenum, Aspergillus flavus, and Aspergillus niger were isolated and displayed multiple antibiotic resistance. The secondary metabolites secreted by co-culture of L. fermentum + P. ostreatus + S. cerevisiae, and co-culture of P. ostreatus + S. cerevisiae have the highest (P ≤ 0.05) zones of inhibition (23.70 mm) and (21.10 mm) against E. coli, respectively. Metabolites from mono-cultured L. fermentum, P. ostreatus, and S. cerevisiae showed zones of inhibition against indicator microorganisms with values ranging from 8.80 to 11.70 mm, 9.00 to 14.30 mm, and 9.30 to 13.00 mm, respectively. Some of the bioactive compounds found in the metabolites of co-cultured microorganisms were alpha-linolenic acid (25.71%), acetic acid 3-methylbutyl ester (13.83%), trans-squalene (12.39%), pentadecylic acid (11.68%), 3- phenyllactic acid (30.13%), linolelaidic acid methyl ester (15.63%), and 4-O-methylmannose (53.74%).ConclusionRTE foods contain multiple antibiotics resistance pathogens. The pronounced antimicrobial activity of the secondary metabolites against microorganisms from RTE foods could be attributed to the presence of bioactive compounds in the metabolites. These metabolites can be exploited as alternative food preservatives, biopharmaceuticals and can be used towards better health delivering systems.

Volatile Organic Compounds from Ceratocystis cacaofunesta, a Causal Agent of Ceratocystis Wilt of Cacao.

Fungi of the genus Ceratocystis are aggressive tree pathogens that cause serious diseases in several crops around the world. Ceratocystis wilt disease caused by C. cacaofunesta has been shown to be responsible for severe reductions in cacao production. In this study, headspace solid-phase microextraction (HS-SPME) coupled with gas chromatography-mass spectrometry (GC-MS) was used in combination with chemometric analysis for monitoring volatile organic compounds (VOCs) released from C. cacaofunesta. Low-molecular-weight esters, alcohols, ketones, and sulphur compounds were identified in the liquid broth. Monitoring the volatile profile over five days of fungal growth revealed that the concentrations of alcohol and esters were inversely proportional. Acetate esters were responsible for the intense fruity aroma of the C. cacaofunesta culture produced within the first hours after fungal inoculation, which decreased over time, and are likely associated with the attraction of insect vectors to maintain the life cycle of the pathogen. PCA revealed that 3-methylbutyl acetate was the metabolite with the highest factor loading for the separation of the VOC samples after 4h of fungal growth, whereas ethanol and 3-methylbutan-1-ol had the highest factor loadings after 96 and 120h. 3-Methylbutan-1-ol is a phytotoxic compound that is likely associated with host cell death since C. cacaofunesta is a necrotrophic fungus. Fungal VOCs play important roles in natural habitats, regulating developmental processes and intra- and interkingdom interactions. This is the first report on the volatiles released by C. cacaofunesta.

Effect of fatty acid composition on the volatile compounds of pasteurized milk during low-temperature storage

The change in milk fat during storage greatly influences its flavor. This study investigates the effect of fatty acid composition on milk flavor by analyzing volatile compounds in pasteurized whole milk (PWM) and pasteurized skim milk (PSM) during storage at 4 °C. 33 types of volatile compounds were detected and the content of ketones was highest, followed by esters and aldehydes. Based on variable importance in projection and relative odor activity value, 2-hexenal dimer, acetic acid ethyl ester dimer, acetic acid ethyl ester, and butanal were identified as the key differential volatile compounds. These compounds were found in higher concentrations in PWM than in PSM, indicating a close relationship with the changes in the fatty acid composition of milk fat. Among 11 fatty acids detected in PWM, the content of saturated fatty acids (SFA) and polyunsaturated fatty acids (PUFA) decreased by 0.69 % and 49.1 %, respectively, while the content of monounsaturated fatty acids increased by 46.8 % during 15 days storage, which suggests that the oxidation of SFA and PUFA contributed more to the volatile compound formation. Correlation analysis between fatty acid composition and volatile compounds found that fatty acid C18:2 and C16:0 were strongly associated for 2-hexenal, acetic acid ethyl ester, and butanal. These fatty acids were mainly derived from neutral lipids or phospholipids. These findings provide a new perspective for the formation pathway of milk flavor.

Two-Stage Screening of Metschnikowia spp. Bioprotective Properties: From Grape Juice to Fermented Must by Saccharomyces cerevisiae.

Gluconobacter oxydans (Go) and Brettanomyces bruxellensis (Bb) are detrimental micro-organisms compromising wine quality through the production of acetic acid and undesirable aromas. Non-Saccharomyces yeasts, like Metschnikowia species, offer a bioprotective approach to control spoilage micro-organisms growth. Antagonist effects of forty-six Metschnikowia strains in a co-culture with Go or Bb in commercial grape juice were assessed. Three profiles were observed against Go: no effect, complete growth inhibition, and intermediate bioprotection. In contrast, Metschnikowia strains exhibited two profiles against Bb: no effect and moderate inhibition. These findings indicate a stronger antagonistic capacity against Go compared to Bb. Four promising Metschnikowia strains were selected and their bioprotective impact was investigated at lower temperatures in Chardonnay must. The antagonistic effect against Go was stronger at 16 °C compared to 20 °C, while no significant impact on Bb growth was observed. The bioprotection impact on Saccharomyces cerevisiae fermentation has been assessed. Metschnikowia strains' presence did not affect the fermentation time, but lowered the fermentation rate of S. cerevisiae. An analysis of central carbon metabolism and volatile organic compounds revealed a strain-dependent enhancement in the production of metabolites, including glycerol, acetate esters, medium-chain fatty acids, and ethyl esters. These findings suggest Metschnikowia species' potential for bioprotection in winemaking and wine quality through targeted strain selection.

Chemical Characterization of Cider Produced in Hardanger—From Juice to Finished Cider

Our investigation delves into the previously uncharted territory of cider composition from Norway. This study aimed to obtain an overview of the qualitative and quantitative compositions of general chemical parameters, polyphenols (individual and total expressed as gallic acids equivalents), selected esters, and selected C6-alcohols in ciders with the PDO label Cider from Hardanger. In total, 45 juice and cider samples from the fermentation process were collected from 10 cider producers in Hardanger in 2019, 2020, and 2021. Individual sugars, acids, ethanol, and 13 individual phenols were quantified using HPLC-UV/RI. Seven ethyl esters of fatty acids, four ethyl esters of branched fatty acids, ten acetate esters, two ethyl esters of hydroxycinnamic acids, and four C6-alcohols were quantified using HS-SPME-GC-MS. For samples of single cultivars (‘Aroma’, ‘Discovery’, ‘Gravenstein’, and ‘Summerred’), the sum of the measured individual polyphenols in the samples ranges, on average, from 79 to 289 mg L−1 (the lowest for ‘Summerred’ and highest for ‘Discovery’ and ‘Gravenstein’). Chlorogenic acid was the most abundant polyphenol in all samples. Ethyl butyrate, ethyl hexanoate, ethyl octanoate, ethyl decanoate, ethyl isobutyrate, ethyl 2-methylbutyrate, isoamyl acetate, and hexanol were present at concentrations above the odour threshold and contributed to the fruity flavour of the Cider from Hardanger.

Mechanistic studies on an isothiourea-catalyzed reaction of an aromatic ester with an imine

Increasing attention is being paid to divergent chemical reactions as a strategy for synthesizing complex organic molecules simply by regulating and adjusting reaction conditions. However, the factors controlling this switchable transformation remain unclear. A systematic study was conducted using density functional theory (DFT) at the M06–2X level to determine the mechanism of an isothiourea-catalyzed divergent chemical reaction of an aryl acetic acid ester with an imine in different solvents (acetonitrile and ethanol). The origins of the stereoselectivity and chemoselectivity of the reaction were determined using noncovalent interaction (NCI) and electrostatic potential surface (EPS) analyses. Lactamization reaction occurs preferentially in acetonitrile, and esterification proceeds preferentially in ethanol. The reaction of an ammonium enolate intermediate with an alkynyl imine preferentially generates the SS-configured isomer. The results of this theoretical study provide insight into the general mechanism of isothiourea-catalyzed reactions in which the key intermediate is ammonium enolate.

Experimental and thermodynamic study of amyl acetate, amyl acetoacetate, and isoamyl acetoacetate in CO2 solvent

Acetate ester compounds find widespread use in various applications such as surface coatings, ink formulations, pharmaceuticals, and adhesives. It is essential to investigate the phase transition behavior of amyl acetate, amyl acetoacetate, and isoamyl acetoacetate in high-pressure supercritical CO2 (SC-CO2). The vapor-liquid equilibria (VLE) of the SC-CO2 + amyl acetate, SC-CO2 + amyl acetoacetate, and SC-CO2 + isoamyl acetoacetate systems were examined at different temperatures (313.2 ≤ T ≤ 393.2 K) and pressures (1.67 ≤ P ≤ 20.76 MPa). The solubility curve of these systems shows Type-I phase behavior, and the critical points of these binary mixtures were observed between the critical properties of the pure components involved in the systems. The solubility of amyl acetate, amyl acetoacetate, and isoamyl acetoacetate in the SC-CO2 + amyl acetate, SC-CO2 + amyl acetoacetate, and SC-CO2 + isoamyl acetoacetate systems increases with increasing temperature at constant pressure. The two-parameter model of Peng-Robinson equation of state along with a mixing rule, accurately correlated the phase transition behavior and critical mixtures curves for all three systems. The binary interaction parameters (BIPs) were adjusted, and the minimum root mean square deviation percentage was identified for all three systems. The calculated error% was found to be within reasonable limits, with values of 4.95 %, 3.93 %, and 4.18 % for SC-CO2 + amyl acetate, SC-CO2 + amyl acetoacetate, and SC-CO2 + isoamyl acetoacetate systems, respectively. Furthermore, the interaction parameters for the SC-CO2 + amyl acetoacetate mixture was found to be temperature-dependent, and the tested linear regression correlation coefficient for the BIPs parameters of (kij) and (ηij) are 0.98533 and 0.99083, respectively. This is the first research study on the phase behavior of acetate ester compounds in SC-CO2 solvents, and the results have a significant impact on industries at different operating conditions.

Cyberlindnera fabianii, an Uncommon Yeast Responsible for Gluten Bread Spoilage.

A single strain of yeast was isolated from industrial gluten bread (GB) purchased from a local supermarket. This strain is responsible for spoilage consisting of white powdery and filamentous colonies due to the fragmentation of hyphae into short lengths (dust-type spots), similar to the spoilage produced by chalk yeasts such as Hyphopichia burtonii, Wickerhamomyces anomalus and Saccharomycopsis fibuligera. The isolated strains were identified initially by traditional methods as Wickerhamomyces anomalus, but with genomic analysis, they were definitively identified as Cyberlindnera fabianii, a rare ascomycetous opportunistic yeast species with low virulence attributes, uncommonly implicated in bread spoilage. However, these results demonstrate that this strain is phenotypically similar to Wi. anomalus. Cy. fabianii grew in GB because of its physicochemical characteristics which included pH 5.34, Aw 0.97 and a moisture of about 50.36. This spoilage was also confirmed by the presence of various compounds typical of yeasts, derived from sugar fermentation and amino acid degradation. These compounds included alcohols (ethanol, 1-propanol, isobutyl alcohol, isoamyl alcohol and n-amyl alcohol), organic acids (acetic and pentanoic acids) and esters (Ethylacetate, n-propil acetate, Ethylbutirrate, Isoamylacetate and Ethylpentanoate), identified in higher concentrations in the spoiled samples than in the unspoiled samples. The concentration of acetic acid was lower only in the spoiled samples, but this effect may be due to the consumption of this compound to produce acetate esters, which predominate in the spoiled samples.

Enhanced production of isobutyl and isoamyl acetate using Yarrowia lipolytica.

Short-chain esters, particularly isobutyl acetate and isoamyl acetate, hold significant industrial value due to their wide-ranging applications in flavors, fragrances, solvents, and biofuels. In this study, we demonstrated the biosynthesis of acetate esters using Yarrowia lipolytica as a host by feeding alcohols to the yeast culture. Initially, we screened for optimal alcohol acyltransferases for ester biosynthesis in Y. lipolytica. Strains of Y. lipolytica expressing atf1 from Saccharomyces cerevisiae, produced 251 or 613 mg/L of isobutyl acetate or of isoamyl acetate, respectively. We found that introducing additional copies of ATF1 enhanced ester production. Furthermore, by increasing the supply of acetyl-CoA and refining the culture conditions, we achieved high production of isoamyl acetate, reaching titers of 3404 mg/L. We expanded our study to include the synthesis of a range of acetate esters, facilitated by enriching the culture medium with various alcohols. This study underscores the versatility and potential of Y. lipolytica in the industrial production of acetate esters.

Aptamer-functionalized triptolide with release controllability as a promising targeted therapy against triple-negative breast cancer

Targeted delivery and precise release of toxins is a prospective strategy for the treatment of triple-negative breast cancer (TNBC), yet the flexibility to incorporate both properties simultaneously remains tremendously challenging in the X-drug conjugate fields. As critical components in conjugates, linkers could flourish in achieving optimal functionalities. Here, we pioneered a pH-hypersensitive tumor-targeting aptamer AS1411-triptolide conjugate (AS-TP) to achieve smart release of the toxin and targeted therapy against TNBC. The multifunctional acetal ester linker in the AS-TP site-specifically blocked triptolide toxicity, quantitatively sustained aptamer targeting, and ensured the circulating stability. Furthermore, the aptamer modification endowed triptolide with favorable water solubility and bioavailability and facilitated endocytosis of conjugated triptolide by TNBC cells in a nucleolin-dependent manner. The integrated superiorities of AS-TP promoted the preferential intra-tumor triptolide accumulation in xenografted TNBC mice and triggered the in-situ triptolide release in the weakly acidic tumor microenvironment, manifesting striking anti-TNBC efficacy and virtually eliminated toxic effects beyond clinical drugs. This study illustrated the therapeutic potential of AS-TP against TNBC and proposed a promising concept for the development of nucleic acid-based targeted anticancer drugs.