Enzymatic synthesis of 2-phenethyl acetate in water catalyzed by an immobilized acyltransferase from Mycobacterium smegmatis

Although water is an ideal green solvent for organic synthesis, it is difficult for most biocatalysts to carry out transesterification reactions in water because of the reversible hydrolysis reaction.

Bactrocera oleae (Rossi) (Diptera: Tephritidae) Response to Different Blends of Olive Fruit Fly-Associated Yeast Volatile Compounds as Attractants

The olive fruit fly, Bactrocera oleae (Rossi) is economically the most important olive pest, causing yield losses in all olive growing areas where is detected. Considering that EU requires the reduction of pesticide use by up to 100% by 2050, more effective non-pesticide lures for B. oleae monitoring and/or controlling are needed. This research was aimed at investigating the attractiveness of different blends of olive fruit fly-associated yeast volatiles toward B. oleae. Three blends of olive fruit fly-associated yeast volatiles: isoamyl alcohol and 2-phenethyl alcohol; isoamyl alcohol, 2-phenethyl alcohol and 2-phenethyl acetate; and isoamyl alcohol, 2-phenethyl acetate and isobutyl acetate were selected and tested on yellow sticky traps for attraction of B. oleae in olive orchard. Results showed that traps containing all tested blends of olive fruit fly-associated yeast volatile compounds, in total, were significantly more attractive to B. oleae and were not significantly attractive to green lacewings, compared to the control. Among them, the most promising was the one containing the blend of isoamyl alcohol, 2-phenethyl acetate and isobutyl acetate because its attractiveness was constantly significant during investigation compared to the others. This blend in the future could lead us to the discovery of a new attractant for the monitoring and/or controlling of B. oleae.

Development of Korean Indigenous Low-Temperature-Tolerant Yeast for Enhancing the Flavor of Yakju

Yakju, a traditional fermented beverage in Korea, is prepared using various raw materials and methods, and, hence, exhibits various characteristics. Low-temperature-fermented yakju can inhibit the growth of undesirable bacteria and is known for its unique flavor and refreshing taste. To increase the production of volatile aromatic compounds in yakju, strains with strong resistance to low temperatures and excellent production of volatile aromatic compounds were screened from indigenous fruits (grape, persimmon, plum, aronia, wild grape) and nuruk in Korea. One Saccharomyces cerevisiae and three non-Saccharomyces strains were finally screened, and yakju was fermented at 15 °C through mono/co-culture. The analysis of volatile aromatic compounds showed that S. cerevisiae W153 produced 1.5 times more isoamyl alcohol than the control strain and reduced the production of 2,3-butanediol by a third. Similarly, a single culture of Pichia kudriavzevii N373 also produced 237.7 mg/L of ethyl acetate, whereas Hanseniaspora vineae G818 produced ~11 times greater levels of 2-phenethyl acetate than the control. Alternatively, Wickerhamomyces anomalus A159 produced 95.88 mg/L of ethyl hexadecanoate. During principal component analysis, we also observed that the co-culture sample exhibited characteristics of both volatile aroma compounds of the single cultured sample of each strain. Our results suggest that yakju with unique properties can be prepared using various non-Saccharomyces strains.

Effects of Different Winemaking Yeasts on the Composition of Aroma-Active Compounds and Flavor of the Fermented Jujube Wine

For the winemaking bioprocess of jujube wine, the selection of optimal starter cultures is one of the major concerns before fermentation. In this study, we investigated the effects of different winemaking yeasts on the composition of aroma-active compounds in the fermented jujube wine and identified the principal components that determine the flavor quality. It showed that the starter winemaking yeasts produced a total of 43 aroma-active compounds, of which esters (e.g., ethyl caprylate, ethyl decanoate, ethyl hexanoate, and phenethyl acetate) contribute more to the wine quality attributes, especially for the improvement of the aroma. Moreover, the composition of aroma-active compounds, for example, the ratio of the content of esters and alcohols, exerts a great impact on the flavor quality of jujube wine. Different starter winemaking yeasts resulted in significant differences in the composition (both species and content) of aroma-active compounds, and thus formed different flavors in the jujube wine. Thus, we propose that screening of a desirable starter winemaking yeast is essential before the fermentation of jujube wine at a large scale, and more considerations should be taken into the resulting composition of aroma-active compounds.

Identification and Functional Analysis of SlitOBP11 From Spodoptera litura

Odorant binding proteins (OBPs) play a key role in the olfactory recognition of insects, whose functions have been extensively studied in adult insects but rarely in larvae. In this study, one OBP (SlitOBP11) with high expression in larval antenna but low expression in adult antenna of Spodoptera litura was screened by RNA-seq and verified by quantitative real-time PCR. Furthermore, the function of SlitOBP11 was explored by analysis of the expression patterns and prokaryotic expression of proteins as well as assays of competitive binding. Competitive binding assay demonstrated that SlitOBP11 had high binding affinity to all four female sex pheromone components, but exhibited almost no binding affinity to plant volatiles except for a low affinity to Phenylacetaldehyde and Phenethyl acetate. Homology modeling and molecular docking implied that the shape of these four sex pheromones were linear, which were appropriate for the binding channel of SlitOBP11 and the amino acid residue Asn99 of SlitOBP11 might play an important role in binding. Taken together, our results indicate that SlitOBP11 may be involved in the perception of female sex pheromones by S. litura larvae, and OBPs in the larvae of S. litura play an important role in the olfactory perception process.

Evaluation of the Effect of Auxiliary Starter Yeasts With Enzyme Activities on Kazak Cheese Quality and Flavor

To investigate the effect of yeasts on Kazak cheese quality and flavor, three isolated yeasts (Kluyveromyces marxianus A2, Pichia kudriavzevii A11, and Pichia fermentans A19) were used to ferment cheeses and designated as StC, LhC, and WcC, respectively. The cheese fermented with a commercial lactic acid starter without adding yeast was used as control named LrC. The results showed that the texture of cheese added with yeasts were more brittle. K. marxianus A2 contributed to the formation of free amino acids and organic acids, especially glutamate and lactic acid. Moreover, K. marxianus A2 provides cheese with onion, oily, and floral aromas. Furthermore, P. kudriavzevii A11 promotes a strong brandy, herbaceous, and onion flavor. Although no significant aroma change was observed in PfC, it promoted the production of acetic acid, isoamyl acetate, and phenethyl acetate. These results indicate that yeasts are important auxiliary starters for cheese production.

Influence of Phenethyl Acetate and Naphthalene Addition before and after Pyrolysis on the Quantitative Analysis of Bio-Oil

The condensation-collection and quantitative analysis of bio-oil limit its component investigation and utilization. In order to find a convenient method for the analysis of bio-oil, the present study conducted an attempt for bio-oil quantitative analysis with the addition of internal standards before pyrolysis. Based on their good thermal stability, phenethyl acetate and naphthalene were selected as standards in the study and experiments were carried out to compare the effects of two added modes (adding into the biowaste before pyrolysis or adding into bio-oil after pyrolysis) on the bio-oil analysis. The results showed that both phenethyl acetate and naphthalene were mainly volatilized under testing conditions, which could be transferred into the oil with the volatile matters during biowaste pyrolysis. Through the co-pyrolysis experiments of the internal standards with lignin and cellulose, almost no interactions were found between the internal standards and such components. Furthermore, adding these standards before pyrolysis hardly affected the properties of noncondensable gas and biochar from the used biowaste samples (tobacco and sawdust waste). Compared with the bio-oil analysis results via traditional methods by adding standards into the bio-oil after pyrolysis, the results regarding the component distribution characteristics of the bio-oil were similar using the proposed method through the addition of standards before pyrolysis. Considering adequate mixing of the added standards (before pyrolysis) in the generated bio-oil, the proposed method could partly help to avoid inaccurate analysis of bio-oil components caused by incomplete collection of the pyrolytic volatiles.

Chemoenzymatic Total Synthesis of (+)-10-Keto-Oxycodone from Phenethyl Acetate

The total synthesis of (+)-10-keto-oxycodone was attained from phenethyl acetate in a stereoselective manner. Absolute stereochemistry was established via enzymatic dihydroxylation of phenethyl acetate with the recombinant strain JM109 (pDTG601A) that furnished the corresponding cis-cyclohexadienediol whose configuration corresponds to the absolute stereochemistry of the ring C of (+)-10-keto-oxycodone. Intramolecular Heck reaction was utilized to establish the quaternary carbon at C-13, along with the dibenzodihydrofuran functionality. The C-14 hydroxyl and C-10 ketone were installed via SmI2-mediated radical cyclization, and oxidation of a benzylic alcohol (obtained from an intermediate nitrate azide), respectively. The synthesis of (+)-10-keto-oxycodone was completed in a total of 14 operations (21 steps) and an overall yield of ~2%. Experimental and spectral data are provided for key intermediates and new compounds.

Characterization of the Key Aroma Compounds in Marselan Wine by Gas Chromatography-Olfactometry, Quantitative Measurements, Aroma Recombination, and Omission Tests

Key odorants of red wine made from the hybrid grapes of Marselan (Vitis vinifera L.) were isolated by solid-phase extraction (SPE) and explored by gas chromatography-olfactometry (GC-O) analysis. Application of aroma extract dilution analysis (AEDA) revealed 43 odor-active compounds, and 31 odorants among them were detected with flavor dilution (FD) factors ranging from 9 to 2187. Comprehensive two-dimensional gas chromatography and time-of-flight mass spectrometry (GC × GC-TOF-MS) were exploited to quantitate the aroma-active compounds with FD ≥9. The identification indicated β-damascenone as having the highest FD factors, followed by eugenol, 2,3-butanedione, citronellol, 4-hydroxy-2,5-dimethyl-3(2H)-furanone, phenethyl acetate, guaiacol, and 2-methoxy-4-vinylphenol. A total of 21 compounds were found to have odor activity values (OAVs) >1.0. Aroma reconstitution validation experiments showed a good similarity of blackberry, green pepper, honey, raspberry, caramel, smoky, and cinnamon aroma attributes between the original Marselan wine and the reconstructed wine. In addition, omission tests were carried out to further determine the contribution of odorants to the overall aroma.