Developing Clostridia as Cell Factories for Short- and Medium-Chain Ester Production

Short- and medium-chain volatile esters with flavors and fruity fragrances, such as ethyl acetate, butyl acetate, and butyl butyrate, are usually value-added in brewing, food, and pharmacy. The esters can be naturally produced by some microorganisms. As ester-forming reactions are increasingly deeply understood, it is possible to produce esters in non-natural but more potential hosts. Clostridia are a group of important industrial microorganisms since they can produce a variety of volatile organic acids and alcohols with high titers, especially butanol and butyric acid through the CoA-dependent carbon chain elongation pathway. This implies sufficient supplies of acyl-CoA, organic acids, and alcohols in cells, which are precursors for ester production. Besides, some Clostridia could utilize lignocellulosic biomass, industrial off-gas, or crude glycerol to produce other branched or straight-chain alcohols and acids. Therefore, Clostridia offer great potential to be engineered to produce short- and medium-chain volatile esters. In the review, the efforts to produce esters from Clostridia via in vitro lipase-mediated catalysis and in vivo alcohol acyltransferase (AAT)-mediated reaction are comprehensively revisited. Besides, the advantageous characteristics of several Clostridia and clostridial consortia for bio-ester production and the driving force of synthetic biology to clostridial chassis development are also discussed. It is believed that synthetic biotechnology should enable the future development of more effective Clostridia for ester production.

Volatile Esters and Fusel Alcohol Concentrations in Beer Optimized by Modulation of Main Fermentation Parameters in an Industrial Plant

Contents of selected volatile esters and fusel alcohols and their relation to the sensory quality of a bottom-fermented lager beer fermented under high-gravity conditions (15.5 °P) were analyzed using response surface methodology (RSM, Box–Behnken design). The influence of various pitching rates (6–10 mln cells/mL), aeration levels (8–12 mgO2/mL), times (4.5–13.5 h) of filling CCTs (cylindroconical fermentation tanks; 3850 hL), and fermentation temperatures (8.5–11.5 °C) on the contents of selected esters, as well as on concentrations of amyl alcohols and on the sum of higher alcohols in beer, was determined in a commercial brewery fermentation plant. Beers produced throughout the experiments met or exceeded all criteria established for a commercial, marketed beer. Statistical analyses of the results revealed that within the studied ranges of process parameters, models with diversified significance described the concentrations of volatiles in beer. The multiple response optimization procedure analyses showed that the values of process parameters that minimized higher alcohols in beer (97.9 mg/L) and maximized its ethyl acetate (22.0 mg/L) and isoamyl acetate (2.09 mg/L) contents, as well as maximized the sensory quality of beer, (66.4 pts) were the following: Pitching rate 10 mln cells per mL; fermentation temperature 11.5 °C; aeration level 8.8 mg/L; and CCT filling time 4.5 h.

Effects of Ethanol Concentration on Oral Aroma Release After Wine Consumption

This paper evaluates, for the first time, the effects of ethanol concentration on the dynamics of oral (immediate and prolonged) aroma release after wine consumption. To do this, the intraoral aroma release of 10 panelists was monitored at two sampling points (0 and 4 min) after they rinsed their mouths with three rosé wines with different ethanol content (0.5% v/v, 5% v/v and 10% v/v) that were aromatized with six fruity esters (ethyl butanoate, isoamyl acetate, ethyl pentanoate, ethyl hexanoate, ethyl octanoate and ethyl decanoate). Overall, the results indicated that the extent of the effects of ethanol content on the oral aroma release were influenced by the subject, the ethanolconcentration and the type of aroma compound. This effect was also different in the immediate than in the prolonged aroma release. In the first in-mouth aroma monitoring, an increase in the ethanol content provoked a higher release of the more polar and volatile esters (ethyl butanoate, ethyl pentanoate), but a lower release for the more apolar and less volatile esters (ethyl octanoate, ethyl decanoate). Regarding the prolonged oral aroma release, an increase of ethanol content in wine increased the oral aroma release of the six esters, which might also increase the fruity aroma persistence in the wines. Future works with a higher number of individuals will be needed to understand the mechanisms behind this phenomenon.