Low-temperature brewing by freeze-dried immobilized cells on gluten pellets

A biocatalyst, prepared by the immobilization of a cryotolerant strain of Saccharomyces cerevisiae on gluten pellets, was freeze-dried without any protecting medium and used for repeated batch fermentations of wort for each of the temperatures 15, 10, 5, and 0 degrees C. The fermentation time for freeze-dried immobilized cells was about 2-fold that of the corresponding time for wet immobilized cells on gluten pellets, and lower than the corresponding time for freeze-dried free cells, especially at 5 and 0 degrees C. Beers produced by freeze-dried immobilized cells contained alcohol levels in the range of 5.0-5.5% v/v, diacetyl concentrations lower than 0.5 mg/L, polyphenol concentrations lower than 145.5 mg/L, and free cell concentrations lower than 3 g/L. As a result, they had a very good clarity after the end of primary fermentation. The amounts of amyl alcohols were lower than 129.1 mg/L and reduced as the temperature was decreased. Ethyl acetate concentrations were found in the range of 22.1-29.2 mg/L, giving a very good aroma and taste in the produced beers.     

Evaluation of the thermally dried immobilized cells of Lactobacillus delbrueckii subsp. bulgaricus on apple pieces as a potent starter culture

The aim of the present study was to evaluate the impact of thermal drying of immobilized Lactobacillus delbrueckii subsp. bulgaricus on apple pieces on the use of the derived biocatalyst in whey fermentation. The thermally dried immobilized biocatalyst was compared to wet and freeze-dried immobilized cells, in respect to maintenance of cell viability and fermentation efficiency. The thermal drying process appeared to be more efficient on survival rate as an 84% of the cells used for immobilization survived the process, while the freeze-drying process led to a 78% rate. The thermally dried immobilized biocatalyst was used in 12 repeated batch fermentations of synthetic lactose medium and whey at 37, 45, and 50 degrees C in order to evaluate its metabolic activity. The high number of repeated batch fermentations showed a tendency for high operational stability. Fermentations continued for up to 2 months without any significant loss of metabolic activity. SPME GC/MS analysis of aroma-related compounds revealed the distinctive character of fermented whey produced by the thermally dried immobilized bacterium cells. The effect of storage at 4-6 degrees C for up to 165 days of the biocatalyst, held directly after drying and after repeated batch fermentations, on fermentation activity was also studied. After storage, reactivation in whey was immediate, and the immobilized biocatalyst was able to produce up to 51.7 g/L lactic acid at 37 degrees C. The potential of thermally dried immobilized L. delbrueckii as a starter culture for food production was subsequently evaluated.     

Volatile compounds of wines produced by cells immobilized on grape skins

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae cells on grape skins. Repeated batch fermentations were conducted using the immobilized biocatalyst as well as the free yeast cells at 25, 20, 15, and 10 degrees C. The major volatile byproducts were determined by GC, whereas the minor volatile constituents were extracted in dichloromethane and analyzed by HRGC-MS. The qualitative profiles of the wines produced were similar in every case. Immobilized cells gave wines with higher contents of ethyl and acetate esters that increased with temperature decreases from 25 to 15 degrees C. The amount of volatile alcohols was more pronounced in wines produced by free cells and decreased dramatically at low fermentation temperatures (10 degrees C).     

Wine production using yeast immobilized on apple pieces at low and room temperatures

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae strain AXAZ-1 on apple pieces. It was examined by electron microscope and studied during the fermentation of grape must for batch wine-making. The immobilized yeast showed an important operational stability without any decrease of its activity even at low temperatures (1-12 degrees C). Specifically, at 6 degrees C the biocatalyst favored wine production within 8 days, which is less time than is required for the natural fermentation of grape must. At 1 degrees C wine production was effected in 1 month. This speeding up of the fermentation could be accepted and adopted by the industry for scaling up the wine-making process. Total and volatile acidities were similar to those found in dry wines. The concentrations of higher alcohols (propanol-1 and isobutyl alcohol) were low. The presence of amyl alcohols proved to be temperature dependent and decreased with the temperature decrease. The values of ethyl acetate concentrations were relatively high, up to 154 mg/L. This probably contributes to the fruity aroma and excellent taste of the produced wines. There was no indication of vinegar odor in the product given that the volatile acidity was <0.47 g of acetic acid/L. From the GC-MS analysis it was concluded that cell immobilization did not create serious changes in the product (wine) with regard to the qualitative composition of the aroma compounds.     

Evolution of volatile byproducts during wine fermentations using immobilized cells on grape skins

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae cells on grape skins. Repeated batch fermentations were conducted using this biocatalyst as well as free cells, at 25, 20, 15, and 10 degrees C. Solid phase microextraction (SPME) was used in monitoring the evolution of volatile byproducts. The effect of immobilization and temperature on evolution patterns of volatiles was obvious. The major part of esters was formed after consumption of 40-50% of the sugars. Similar processes were observed for amyl alcohols and 2-phenylethanol, whereas 1-propanol and 2-methyl-1-propanol were formed during the whole alcoholic fermentation period at an almost constant formation rate. Acetaldehyde and acetoin were synthesized in the early stages of fermentation. Afterward, their amount decreased. In most cases, immobilized cells exhibited higher formation rates of volatiles than free cells. The final concentration of esters was higher in wines produced by immobilized biocatalyst. Their amount increased with temperature decrease. The opposite was observed for higher alcohols.     

Interesterification of olive oil with a fully hydrogenated fat in a batch reactor using step changes in temperature

Interesterification of a 60:40 (wt/wt) mixture of olive oil and fully hydrogenated canola oil was carried out in a batch reactor using a commercial immobilized lipase from Thermomyces lanuginose as a biocatalyst. The effects of a stepwise change of temperature on the degree of conversion, the solid fat content (SFC) of the products, and the residual activity of the enzyme were investigated. As a reference condition, an interesterification trial was conducted at a constant temperature of 70 degrees C for 48 h. For trials in which a temperature of 70 degrees C was used for the first 4 h of reaction and a temperature of 60 degrees C was employed for the following 44 h, there were no significant differences (p < 0.05) in the overall degree of conversion relative to the reference condition. Oils interesterified for only 1 or 2 h at 70 degrees C had melting points higher than 60 degrees C, whereas an oil produced by interesterification at 70 degrees C for only 4 h had a melting point of 58 degrees C. There was little difference (p < 0.05) between the SFC profiles of the interesterification products prepared by two different temperature protocols (70 degrees C for 24 h; 70 degrees C for 4 h followed by 60 degrees C for 20 h). Use of the protocol involving a step decrease in temperature significantly decreased catalyst deactivation effects, thereby increasing the residual activity of the immobilized lipase.     

Effect of carbohydrate substrate on fermentation by kefir yeast supported on delignified cellulosic materials

The suitability of delignified cellulosic (DC) material supported kefir yeast to ferment raw materials that contain various single carbohydrates, for the production of potable alcohol and alcoholic drinks, is examined in this investigation. Results are reported of fermentations carried out with sucrose, fructose, and glucose in synthetic media. Repeated batch fermentations at various initial sugar concentrations of sucrose, fructose, and glucose were performed at 30 degrees C in the presence of the aforementioned biocatalyst. The results clearly show feasible yields in the range of 0.38-0.41 g/g, alcohol concentrations of 7.6-8.2% v/v, fermentation time of 90-115 h, and conversion of 92-96%. DC material supported kefir fermented 11-fold more rapidly than free cells and 9-fold more rapidly in comparison to kissiris supported kefir. The main volatile byproducts such as amyl alcohols (mixture of 2-methyl-1-butanol and 3-methyl-1-butanol), ethanal, and ethyl acetate were formed in all sugar fermentation products. The formation of 65-110 ppm of ethyl acetate is as high and even higher than that obtained with traditional wine yeasts. The increase of the initial concentration of sugar in the fermentation media resulted in an increase in contents of volatiles. The fine aroma that was obtained in the product of fructose could be attributed to the high percentage of ethyl acetate on total volatiles. The efficiency of DC material supported kefir was the same for the fermentations of individual sugars or a mixture of fructose, sucrose, and glucose. When whey with raisin extracts was fermented, lower yields were obtained but the aroma of the product was even better.     

Food additive lactic acid production by immobilized cells of Lactobacillus brevis on delignified cellulosic material

Improvements in yield and productivity in lactic acid fermentation by Lactobaccilus brevis cells immobilized on delignified cellulosic (DC) material are reported. The system proved to be more efficient in comparison with the work reported by other workers. Yields of 80 and 100% conversion using glucose were obtained at 30 degrees C in 1 day of fermentation time. Lactic acid fermentation using whey as substrate was obtained at 30 degrees C in 1-1.5 days, resulting in 70% yield, whereas the remaining lactose in whey was converted to alcohol byproduct, leading to a 90% lactose exploitation and 100% conversion. Cell immobilization of L. brevis on DC material was proved by its reuses in repeated batch fermentations and through electron microscopy. A series of 10 repeated batch fermentations without any loss in cell activity showed a tendency for high operational stability. The presence of DC material resulted in a drastic drop of the fermentation time from 48 to 13 h.     

Investigation of volatiles evolution during the alcoholic fermentation of grape must using free and immobilized cells with the help of solid phase microextraction (SPME) headspace sampling

A biocatalyst was prepared by immobilization of Saccharomyces cerevisiae strain AXAZ-1 on delignified cellulosic material (DCM). Repeated batch fermentations were conducted using these biocatalysts and free cells, separately, at temperatures of 20, 15, and 10 degrees C. Solid phase microextraction (SPME) was used in monitoring the formation of volatile alcohols, acetate esters, and ethyl esters of fatty acids. The kinetics of volatile production were similar for free and immobilized cells. In all cases immobilized cells showed a better rate of volatile production, which was directly connected to sugar consumption. The main difference observed was in propanol production, which increased with temperature decrease for the immobilized cells, whereas it remained constant for the free ones. In the case of immobilized cells significant amounts of esters were also produced. It is well-known that esters contribute to the fruity aroma of wine. It was also established that SPME is a very sensitive, accurate, and reliable technique and can be used without any reservation in the characterization of volatile constituents of wine.     

Delignified cellulosic material supported biocatalyst as freeze-dried product in alcoholic fermentation

Freeze-dried delignified cellulosic (DC) material supported biocatalyst is proposed as a suitable form of biocatalyst to be preserved. The alcoholic fermentation of glucose using freeze-dried immobilized cells is reported. Freeze-dried immobilized baker's yeast cells on DC material do not need any protective medium during freeze-drying. The effect of initial glucose concentration and temperature on the alcoholic fermentation kinetic parameters is reported in the present study. It was found that the freeze-dried immobilized cells ferment more quickly than free freeze-dried cells and have a lower fermentation rate as compared with wet immobilized cells. However, repeated batch fermentations showed freeze-dried immobilized cells to ferment at about the same fermentation rate as wet immobilized cells. The results indicate that the freeze-dried immobilized cells must be further studied to establish a process for the preservation of immobilized cells.     

Whey liquid waste of the dairy industry as raw material for potable alcohol production by kefir granules

Kefir granular biomass was used in the fermentation of sweet whey and proved to be more effective compared to single-cell biomass of kefir yeast. The operational stability of the biocatalyst was assessed by carrying out 20 repeated batch fermentations. Levels of ethanol productivity reached 2.57 g L(-1) h(-1)), whereas the yield was 0.45 g/g. The fermentation time was only 8 h. Mixtures of sweet whey with molasses were fermented at initial densities ranging from 4.2 to 10.2 degrees Be and resulted in ethanol yield factors between 0.36 and 0.48 g of ethanol/g of utilized sugar. Lower degrees Be values led to an increase of percentages of ethyl acetate on total volatiles determined and a reduction of amyl alcohols. The addition of 1% black raisin extract to whey appears to promote whey fermentation, whereas the same was not observed in the case of white sultana extract addition. It was finally established that it is preferable to ferment mixtures of whey-molasses by adding molasses in whey after the completion of whey fermentation.     

Red wine making by immobilized cells and influence on volatile composition

Red wine making using yeast cells immobilized in two types of raisin berries, at various temperatures (6-30 degrees C), was studied. A modification of the batch bioreactor was used to separate the grape skins used for color extraction from the biocatalyst and the fermenting grape must. The evaluation of the immobilized biocatalysts was made on terms of productivity and organoleptic quality, including color intensity and formation of volatiles. The immobilized cells were found capable of low-temperature wine making, producing red wines containing more than 11% v/v alcohol in 8 days at 6 degrees C. The quality of wines was examined by gas chromatography (GC) and GC-MS analysis and sensory evaluation. Higher alcohol concentrations were decreased, and ethyl acetate concentrations increased by the drop of temperature. Many esters, alcohols, carbonyls, and miscellaneous compounds were identified in wines produced by immobilized cells, revealing no significant qualitative differences as compared to wines produced by free cells. The sensory evaluation showed that the best red wine was produced at 6 degrees C.     

Extremely low temperature fermentations of grape must by potato-supported yeast, strain AXAZ-1. A contribution is performed for catalysis of alcoholic fermentation

This investigation announces the use of potato pieces as a suitable support for cell immobilization resulting in extremely low temperature wine making. The results showed an increase of the total esters by immobilized cells and reduction of higher alcohols. Likewise, percentages of total esters on total volatiles were increased by the drop in temperature, while percentages of higher alcohols were reduced in wines. Kinetics experiments at different temperatures allowed the calculation of activation energy (Ea) and showed reduction in the case of immobilized cells as compared with free cells. These results may lead to the conclusion that the increased productivities that are obtained by immobilized cells, can be attributed to the catalytic activity by the support to enzymes, which are involved in the process. Biocatalysts were prepared by immobilization of Saccharomyces cerevisiae, strain AXAZ-1, on whole potatoes and potato pieces, and their efficiency for alcoholic repeated batch fermentations of glucose and grape must in the range 2-30 degrees C was examined. To study the operational stability of biocatalyst, 35 repeated batch fermentations of grape must were performed without any significant reduction of the fermentation activity. Wines were analyzed for volatile byproducts determination by GC and GC-MS.     

Storage of immobilized yeast cells for use in wine-making at ambient temperature

A comparative study of the storage and reuse of immobilized yeast cells on apple pieces, kissiris, and gamma-alumina was carried out. The immobilized biocatalysts were allowed to remain in the fermented alcoholic liquid after the end of each fermentation batch for extended periods at 30 degrees C before reactivation in batch fermentation for wine-making. The results showed that the biocatalysts were able to reactivate and ferment after successively increased periods of storage compared to free cell systems both on glucose medium and on grape must. In glucose medium, apple-, kissiris-, and gamma-alumina-supported biocatalysts reactivated after 120, 80, and 83 days, respectively. Possible storage periods for grape must were lower but remained high. Immobilized yeast biocatalyst on apple pieces produced wines with an improved volatiles composition compared to kissiris- and gamma-alumina-supported biocatalysts. There were no significant negative effects on the fermentation activity and volatile byproduct composition.     

Study of some Saccharomyces cerevisiae strains for winemaking after preadaptation at low temperatures

Low-temperature fermentations (13 degrees C) are considered to improve wine aromatic profiles. However, because the risk of stuck and sluggish fermentations is high, these fermentations are not common. The aim of this paper was to analyze the effect of different preadaptation protocols in two commercial wine strains on the fermentation and some wine parameters. Preadaptation is understood to be the process between the rehydration of active dry yeast and the inoculation. In this study, it consisted of preparing a fermentation starter (addition of yeast grown at 25 degrees C) or inocula preadapted at low temperatures (as before, but grown at a fermentation temperature of 13 or 17 degrees C). These results were compared with those of rehydrated active dry yeast, and a commercial "cryotolerant" yeast was used as a reference. General fermentation kinetic parameters, yeast imposition, nitrogen consumption, and main wine products were analyzed. The results showed that the preadaptation of a yeast could improve the fermentation performance, although this improvement was strain-dependent. Low-temperature fermentations also had some general effects: reduction of acetic acid and fusel alcohol production and increased concentrations of glycerol. When the yeast performed better in fermentation because of preadaptation, nitrogen consumption was faster and the wine's "negative" attributes (acetic acid, fusel alcohols) were significantly reduced. Thus, in some strains, preadaptation could be an effective mechanism for improving low-temperature fermentation, which also significantly reduces detrimental wine attributes.     

Continuous whey fermentation using kefir yeast immobilized on delignified cellulosic material

Delignified cellulosic-supported biocatalyst, prepared by immobilization of kefir yeast on delignified cellulosic material (DCM), was found to be suitable for continuous, modified whey fermentation. The modified whey contained 1% raisin extract and molasses. Ethanol productivities ranged from 3.6 to 8.3 g L(-1)day(-1), whereas parameters such as ethanol concentration, residual sugars, and daily fermented whey productivity were acceptable for the production of potable alcohol and alcoholic drinks in industrial fermentations. The continuous fermentation bioreactor was operated for 39 days, stored for 18 days at 4 degrees C, and operated again for another 15 days without any diminution of the ethanol productivity. The concentrations of higher alcohols (propanol-1, isobutyl alcohol, and amyl alcohols) were low. The main volatile byproducts formed in the continuous process were similar to those observed in alcoholic beverages, and the fermented whey had a good aroma. The concentrations of higher alcohols were very low when compared to that of ethyl acetate, therefore resulting in a quality product. The possibility of using such a process for the production of potable alcohol or a novel, low-alcohol content drink is proposed.     

Malolactic fermentation in wine with Lactobacillus casei cells immobilized on Delignified cellulosic material

In this work Lactobacillus casei ATCC 393 cells immobilized on delignified cellulosic material (DCM) were used for malolactic fermentation (MLF) of wine. Wine was produced using yeast cells immobilized on DCM at 20 degrees C, and after alcoholic fermentation, MLF at 27 degrees C followed using immobilized L. casei ATCC 393 cells. A total of 11 repeated alcoholic and subsequent MLF batches were performed within a period of 1 month. As the repeated MLF batches proceeded, the MLF activity of the immobilized biocatalyst was reduced. Malic acid degradation was reduced from 80 to 2%, pH was reduced by 0.5-0.1 unit, acetic acid concentrations were slightly reduced or remained stable (0.002 g/L), the higher alcohols 1-propanol, isobutyl alcohol, and amyl alcohol were decreased by 84, 23, and 11%, respectively, and ethyl acetate concentration was increased by approximately 56%. Wine samples were analyzed by GC-MS before and after MLF, revealing some qualitative differences.     

戴尔有孢圆酵母调控晚采小芒森葡萄酒乙酸和香气

为降低甜型葡萄酒中的挥发酸,进一步增加其香气复杂度,该研究利用本土戴尔有孢圆酵母R12与酿酒酵母NX11424同时接种和顺序接种发酵,研究其对贺兰山东麓产区晚采小芒森葡萄酒乙酸及香气成分的调控。与酿酒酵母单菌发酵相比,本土戴尔有孢圆酵母与酿酒酵母按照5:1、20:1、50:1的不同菌体数量比例同时接种和间隔5d顺序接种发酵,均可以显著降低晚采小芒森葡萄酒的乙酸含量,其乙酸产率分别降低19.1%、21.2%、38.2%和48.9%。此外,按20:1比例同时接种发酵,可显著提高晚采小芒森葡萄酒中萜烯类和降异戊二烯类等品种香气物质含量,和乙酸异戊酯、丁酸乙酯及苯乙醇等发酵香气物质的含量。该研究表明合理的酿酒酵母接种发酵能有效降低高糖原料酿造葡萄酒挥发酸含量,为贺兰山东麓产区葡萄酒新产品的开发提供了的技术参考。     

Investigation of the beta-damascenone level in fresh and aged commercial beers

This study investigated the increase of beta-damascenone content during aging in a variety of commercial Belgian beers. Quantities detected in fresh beers were generally low (from 6 ng/g to 25 ng/g). After 5 days at 40 degrees C, the level increased (to as much as 210 ng/g) in most of the beers studied, according to the type of beer. Further experiments showed that wort initially contains large quantities of beta-damascenone (450 ng/g), but that degradation of the compound during fermentation accounts for the low concentrations observed in fresh beers. Production during beer aging can be partially explained by acid hydrolysis of glycosides.     

Digestion residues of typical and high-beta-glucan oat flours provide substrates for in vitro fermentation

In vitro fermentabilities of the oat flour digestion residues (ODR) from two commercial oat lines with 4.7 and 5.3% beta-glucan and from two high-beta-glucan experimental lines with 7.6 and 8.1% beta-glucan were evaluated and compared with fermentations of lactulose, purified oat beta-glucan (POBG), and purified oat starch (POS). Substrates were fermented by using an in vitro batch fermentation system under anaerobic conditions for 24 h. The progress of the fermentation was studied by following the change in pH of the fermentation medium, production of short-chain fatty acids (SCFA) and gases, and consumption of carbohydrates. The substrate from the flour with the greatest amount of beta-glucan tended to have the greatest pH decline and the greatest total SCFA production. A significant correlation occurred between gas production and SCFA formation (R 2 = 0.89-0.99). Acetate was produced in the greatest amounts by all of the substrates except POBG, by which butyrate was produced in the greatest amount. More propionate and butyrate, but less acetate, were produced from high-beta-glucan ODR. With the given fermentation conditions, >80% of the total carbohydrate was depleted by the bacteria after 24 h. Glucose was the most rapidly consumed carbohydrate among other available monosaccharides in the fermentation medium. Overall, the high-beta-glucan experimental lines provided the best conditions for optimal in vitro gut fermentations.