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Glass transition temperatures and fermentative activity of heat-treated commercial active dry yeasts.

Differential scanning calorimetry thermograms of various samples of commercial instant active dry yeasts revealed a clear glass transition typical of amorphous carbohydrates and sugars. The resulting glass transition temperatures were found to decrease with increasing moisture content. The observed glass curve was similar to that of pure trehalose, which is known to accumulate in large amounts in baker's yeast. The effect of heat treatment at various temperatures on the fermentative activity (as measured by the metabolic production of CO(2)) of dry yeast was studied. First-order plots were obtained representing the loss of fermentative activity as a function of heating time at the various temperatures assayed. Significant losses of fermentative activity were observed in vitrified yeast samples. The dependence of rate constants with temperature was found to follow Arrhenius behavior. The relationship between the loss of fermentative activity and glass transition was not verified, and the glass transition was not reflected on the temperature dependence of fermentative activity loss.

Biotechnology↗

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.

Animals↗

Effect of fermentation on Sweetpotato (Ipomoea batatas) toxicity in mice.

Unfortunate bovine fatalities occurring after ingestion of mold-damaged sweetpotatoes preclude the use of the culled tubers in livestock feed. In cattle, mold-damaged sweetpotatoes induce an acute respiratory distress syndrome resulting in asphyxiation. Because of this potential toxicity and the general abundance of culled sweetpotatoes, the detoxification efficacy of ensiling was explored since it is an easy and economically viable technique often applied to preserve livestock feed. Sweetpotato slices with or without mold damage were stored either frozen (to represent unfermented samples) or fermented for 6 weeks at room temperature. Following fermentation, organic extracts were generated for administration to mice. Thirty hours following administration of the extracts, mice were evaluated for gross and microscopic lesions affecting the lungs, liver, and kidneys. Fermentation of 6 weeks duration was observed to inadequately eliminate the lung, liver, and kidney toxicity caused by mold-damaged sweetpotatoes. In fact, fermentation exacerbated the hepatotoxicity of mold-damaged sweetpotatoes. This is also the first demonstration that sweetpotato regions lacking visible mold damage can induce lung and kidney injury, which, however, is preventable by fermentation.

Animals↗

Influence of thermally processed carbohydrate/amino acid mixtures on the fermentation by Saccharomyces cerevisiae.

The production of alcoholic beverages such as Tequila, Mezcal, whiskey, or beer includes the fermentation of a mash containing Maillard reaction products. Because excessive heating of the mash can lead to complications during the following fermentation step, the impact of Maillard products on the metabolism of Saccharomyces cerevisiae was investigated. For this purpose, fermentation was carried out in a model system in the presence and absence of Maillard reaction products and formation of ethanol served as a marker for the progression of fermentation. We found that increasing amounts of Maillard products reduced the formation of ethanol up to 80%. This effect was dependent on the pH value during the Maillard reaction, reaction time, as well as the carbohydrate and amino acid component used for the generation of Maillard reaction products. Another important factor is the pH value during fermentation: The inhibitory effect of Maillard products was not detectable at a pH of 4 and increased with higher pH-values. These findings might be of relevance for the production of above-mentioned beverages.

Alcoholic Beverages↗

Glucosinolate derivatives in stored fermented cabbage.

The research focused on the glucosinolate (GLS) breakdown products formed during the fermentation of cabbage. A relationship between the contents of degradation products in fermented cabbage and native GLS in raw cabbage was investigated. The effect of fermented cabbage storage on the contents of individual compounds was also assayed. Ascorbigen formed from one of the degradation products of glucobrassicin (indole GLS) was found to be a dominating compound in fermented cabbage. Irrespective of the time of fermented cabbage storage, the content of ascorbigen reached approximately 14 micromol/100 g. Neither the content of isothiocyanates, the major degradation products of aliphatic GLS, nor that of cyanides exceeded 2.5 microM. Storage of cabbage caused periodical increases and decreases in the contents of cyanides and consequent declines in the contents of isothiocyanates. The highest relative contents (expressed as a percentage of the native GLS content) of degradation products--ranging from >70 to 96%--were reported for the products of glucoraphanin degradation, whereas the lowest-- <5% --were reported for the products of sinigrin degradation.

Ascorbic Acid↗

Fermentation of plant cell wall derived polysaccharides and their corresponding oligosaccharides by intestinal bacteria.

New types of nondigestible oligosaccharides were produced from plant cell wall polysaccharides, and the fermentation of these oligosaccharides and their parental polysaccharides by relevant individual intestinal species of bacteria was studied. Oligosaccharides were produced from soy arabinogalactan, sugar beet arabinan, wheat flour arabinoxylan, polygalacturonan, and rhamnogalacturonan fraction from apple. All of the tested substrates were fermented to some extent by one or more of the individual species of bacteria tested. Bacteroides spp. are able to utilize plant cell wall derived oligosaccharides besides their reported activity toward plant polysaccharides. Bifidobacterium spp. are also able to utilize the rather complex plant cell wall derived oligosaccharides in addition to the bifidogenic fructooligosaccharides. Clostridium spp., Klebsiella spp., and Escherichia coli fermented some of the selected substrates in vitro. These studies do not allow prediction of the fermentation in vivo but give valuable information on the fermentative capability of the tested intestinal strains.

Bacteria↗

Influence of a Saccharomyces cerevisiae selected strain in the volatile composition of rosé wines. Evolution during fermentation.

There has been considerable controversy about the use of selected pure strains in wine fermentation. For that reason it is important to determine the influence of this vinification technique in the composition of wine because it arises from the type of yeast and the subsequent evolution during fermentation. This study researches the volatile composition of rosé wines from the Garnacha must, inoculated with one selected NA33 strain of Saccharomyces cerevisiae. The inoculated yeast did not predominate in all of the samples. These samples showed a behavior intermediate between those of the control and samples in which NA33 did predominate. The greatest concentration of higher alcohols was in the control wine, and its evolution was similar in all fermentations. The esters formed at the end of the fermentation and their concentrations were higher in the control than in the inoculated samples. In the control, acids were produced above all, in the first half of fermentation, and decreased from then onward. In the sample in which the yeast predominated, the synthesis occurred later and to a lesser extent than in the control.

Biological Evolution↗

Product and redox potential analysis of sauerkraut fermentation.

The relationships between the redox potential of the brine, during fermentation of white cabbage into sauerkraut of two early and two late fermentation processes, and the changes in the amount of sugars, organic acids, the redox potential of the brine and of the ascorbic acid redox couple, and pH are described. The trend in the change of the redox potential of the brine is the same for all four fermentation processes studied. In the first phase a sharp decrease in redox potential is followed by an increase in redox potential. In the second phase the redox potential is rather constant. This second phase is followed by another decrease in redox potential, which stabilizes at a minimum value, the third phase. It was observed that sugar fermentation and acid production mainly took place during the first and third phases, probably representing, respectively, the heterogeneous and homogeneous fermentation processes.

Ascorbic Acid↗

Aerobic fermentation during tobacco pollen development.

In vegetative organs of plants, the metabolic switch from respiration to fermentation is dictated by oxygen availability. The two genes dedicated to ethanolic fermentation, pyruvate decarboxylase and alcohol dehydrogenase, are induced by oxygen deprivation and the gene products are active under oxygen stress. In pollen, these two genes are expressed in a stage-specific manner and transcripts accumulate to high levels, irrespective of oxygen availability. We have examined the expression pattern of pyruvate decarboxylase and alcohol dehydrogenase at the protein level in developing pollen and show that the active proteins are localized to the gametophytic tissue and begin to accumulate at microspore mitosis. A flux through the ethanolic fermentation pathway could already be detected very early in pollen development, occurring in all stages from premeiotic buds to mature pollen. This flux was primarily controlled not by oxygen availability, but rather by sugar supply. At a high rate of sugar metabolism, respiration and fermentation took place concurrently in developing and germinating pollen. We propose that aerobic fermentation provides a shunt from pyruvate to acetyl-CoA to accommodate the increased demand for energy and biosynthetic intermediates during pollen development and germination. A possible undesirable side-effect is the potential accumulation of toxic acetaldehyde. Our results support a model for cms-T-type male sterility in maize, in which degeneration of the tapetum is caused by the toxic effects of acetaldehyde on mitochondria weakened by the presence of the URF13 protein.

Aerobiosis↗

Effect of blanching time and salt concentration on pectolytic enzymes, texture and acceptability of fermented green beans.

Effect of blanching time, salt concentration and fermentation on texture of green beans (var. Tuf) was determined. Beans were blanched at 90 degrees C for 1, 10 or 20 minutes and fermented with brine containing 3.08 and 4.6% salt (corresponding to 1.2 and 1.8% salt in the finished product). The beans were assessed for texture mechanically using Universal Testing Machine (Model 1011, Instron). Sensory evaluation was carried out to assess the taste, texture and overall acceptability of the products. Pectolytic enzyme (pectinestarase and exopolygalacturonase) activities were determined in fresh and fermented beans. Increase in blanching time improved the texture significantly. Significant (p<0.05) decreases in shearing energy of the beans were observed in the first two days of fermentation and, thereafter, there were no significant (p>0.05) changes except for samples blanched for one minute. Salt concentration showed a small but significant effect on texture only in samples blanched for one minute and this was probably due to activation by salt of residual pectolytic enzymes in the beans. Most acceptable beans were those blanched for 20 minutes and fermented with 1.2% salt brine.

Carboxylic Ester Hydrolases↗

Effect of varied fermentation periods on the diabetogenic potential of toasted cassava granules.

The effects of exclusive gari diets prepared using varied fermentation periods on the fasting blood glucose levels (FBGL) and glucose tolerance (GT) of rats were studied. Thirty growing male albino rats divided randomly into 5 groups of 6 rats each were used. The 5 groups were each fed gari diets fermented for 0, 24, 48, 72 hours; and a standard rat diet, respectively, for 8 weeks. Data on the total cyanogen content and % crude protein of the diets; FBGL, GT, body weight, water:feed consumption ratio and clinical observations of the rats were collected. Results showed that the total cyanogen content and % crude protein of the diets were depleted as fermentation periods increased. The FBGL of all the gari-fed rats were significantly elevated and their GT significantly impaired with a significant variation between the groups. The elevated FBGL and impaired GT which were found to increase with increase in fermentation period were strongly inversely correlated to the % crude protein content of the diets (r = -0.92 and -0.96 respectively) suggesting that the diabetogenic potential of gari diets strongly depended on its % crude protein content. Shortened fermentation periods leading to production of gari with high total cyanogen content did not induce higher diabetogenicity.

Animals↗

A new method for yeast recovery in batch ethanol fermentations: filter aid filtration followed by separation of yeast from filter aid using hydrocyclones.

In the Melle-Boinot process for alcohol production, centrifuges are normally used for yeast recovery at the end of a batch fermentation. Centrifuges are expensive equipment and represent an impressive part of the equipment costs in alcohol industries. In the present work, an alternative method for yeast recovery using less expensive equipment was studied. Instead of using centrifuges, yeast was separated from the fermented broth by filter aid filtration, followed by separation of yeast from the filter aid using hydrocyclones. A stainless steel plate-and-frame filter of filtration area 1.14 m2 and two 30 mm hydrocyclones, which followed the Bradley and Rietema recommended proportions, were used in this work. The filter aid was perlite. Tests of direct separation of yeast from the fermented broth using the Bradley hydrocyclone proved to be completely unfeasible, since the maximal reduced total efficiency obtained was only 1%. When the hydrocyclones were used to separate perlite from the resuspended filtration cake, the perlite total separation efficiency obtained in the underflow was as high as 95% when using the Bradley hydrocyclone with an underflow diameter of 3 mm. To show the feasibility of the proposed new method of yeast recovery, a complete cycle of experiments, which included fermentation, yeast separation, and new fermentation using the recycled cells, was performed with good results.

Centrifugation↗

Development of an owoh-type product from African yam beans (Sphenostylis stenocarpa) (Hoechst (ex. A. Rich.) Harms.) seeds by solid substrate fermentation.

African yam beans were fermented to obtain an owoh-type product. Microorganisms associated with the fermentation were Bacillus licheniformis, B. pumilus, B. subtilis and Staphylococcus sp. Total microbial counts increased from 1.53 x 10(5) cfu/g to 1.51 x 10(9) cfu/g under aerobic conditions, and from 8.0 x 10(3) cfu/g to 1.35 x 10(7) cfu/g under conditions of reduced oxygen tension. The pH of the substrate increased throughout the fermentation, from 6.8 to 7.5. A comparison of unfermented seeds with the fermented product showed that there were decreases in the levels of total nitrogen, crude protein, crude fiber and lipids, and that there were increases in the levels of carbohydrate and total organic matter. Enzyme activities during fermentation revealed that amylase production was erratic showing a slight increase during the first 24 h followed by a steep rise in activity in the next 24 h. By contrast, lipase activity increased rapidly throughout the first 72 h while proteinase activity followed a type of sigmoid curve with a steady increase in activity within the first 48 h and a relatively high activity until the 96th h before plunging downwards.

Amylases↗

Acidogenic fermentation of source separated mixtures of vegetables and fruits wasted from supermarkets.

A pilot scale mesophilic anaerobic acidogenic fermenter was fed with mixtures of vegetables and fruits shredded by a hammer mill and mixed in a stock tank, in order to produce a liquid phase suitable as RBCOD source in denitrification and EBPR processes. Different operative conditions were studied working with a HRT in the range 1-12 days. The effluent coming from the fermenter was screw pressed, and the solid phase was recycled adopting different ratios to the fermenter, in order to define its effect on the final liquid phase composition. The variations of the VFA, lactate, methyl and ethyl alcohol concentrations, TCOD, SCOD and pH during more than one year were analysed and discussed both with reference to the fresh feed, and to the content of the fermenter. It was found that almost all the organic matter in the liquid phase inside the fermenter was represented by VFA (mainly acetate), lactate (in particular) and methyl and ethyl alcohols when HRT was longer than 6 days.

Acids↗

Fermentability of an enzymatically modified solubilised potato polysaccharide (SPP).

OBJECTIVE: To study processing and fermentability in the human gastro-intestinal tract of a newly isolated, enzymatically modified, soluble, and highly concentrated (> 80% dietary fibres) solubilised potato fibre (SPP). SETTING: Gastroenterological laboratory. DESIGN, SUBJECTS AND INTERVENTIONS: Seven healthy volunteers ingested in random order on seven different days: 20 g SPP; bread made of 180 g wheat flour served with 20 g raw SPP; bread baked of 180 g wheat flour and 20 g SPP; bread made from 180 g what flour; 20 g lactulose; 20 g oat bran; and 20 g wheat bran. The hydrogen breath test was used to evaluate oro-coecal transit time (OCTT) and fermentation. RESULTS: Fermentation of SPP yielded a measurable increase in end-expiratory H2. The total incremental increase in end expiratory H2 due to SPP was unaffected of whether SPP was served alone, as the raw flour served with bread, or baked into bread. The OCTT for raw SPP was significantly delayed compared to lactulose (P = 0.01). The OCTT for SPP baked into bread was significantly delayed compared to raw SPP (P = 0.01), indicating that SPP may be used as a marker of oro-coecal transit time for as well the fluid phase as the solid phase of a meal. CONCLUSIONS: SPP is a fermentable, highly concentrated soluble fibre source. Baking SPP did not interfere with the fermentable properties. Thus, SPP may be interesting as a fibre-supplement in fibre-poor diets. The change in oro-coecal transit time for SPP, depending on the composition of the total meal, makes SPP interesting as a marker in studies of gastrointestinal transit.

Avena↗

Maltotriose fermentation by Saccharomyces cerevisiae.

Maltotriose, the second most abundant sugar of brewer's wort, is not fermented but is respired by several industrial yeast strains. We have isolated a strain capable of growing on a medium containing maltotriose and the respiratory inhibitor, antimycin A. This strain produced equivalent amounts of ethanol from 20 g l(-1) glucose, maltose, or maltotriose. We performed a detailed analysis of the rates of active transport and intracellular hydrolysis of maltotriose by this strain, and by a strain that does not ferment this sugar. The kinetics of sugar hydrolysis by both strains was similar, and our results also indicated that yeast cells do not synthesize a maltotriose-specific alpha-glucosidase. However, when considering active sugar transport, a different pattern was observed. The maltotriose-fermenting strain showed the same rate of active maltose or maltotriose transport, while the strain that could not ferment maltotriose showed a lower rate of maltotriose transport when compared with the rates of active maltose transport. Thus, our results revealed that transport across the plasma membrane, and not intracellular hydrolysis, is the rate-limiting step for the fermentation of maltotriose by these Saccharomyces cerevisiae cells.

Beer↗

Succinate production in dual-phase Escherichia coli fermentations depends on the time of transition from aerobic to anaerobic conditions.

We examined succinic acid production in Escherichia coli AFP111 using dual-phase fermentations, which comprise an initial aerobic growth phase followed by an anaerobic production phase. AFP111 has mutations in the pfl, ldhA, and ptsG genes, and we additionally transformed this strain with the pyc gene (AFP111/pTrc99A-pyc) to provide metabolic flexibility at the pyruvate node. Aerobic fermentations with these two strains were completed to catalog physiological states during aerobic growth that might influence succinate generation in the anaerobic phase. Activities of six key enzymes were also determined for these aerobic fermentations. From these results, six transition times based on physiological states were selected for studying dual-phase fermentations. The final succinate yield and productivity depend greatly on the physiological state of the cells at the time of transition. Using the best transition time, fermentations achieved a final succinic acid concentration of 99.2 g/l with an overall yield of 110% and productivity of 1.3 g/l h.

Aerobiosis↗

A continuous fermentation technique for studying the kinetics of sugar uptake by baker's yeast.

1. Solutions of glucose, maltose or other sugars were pumped at controlled rates into a yeast suspension and the extracellular sugar concentration was determined. The technique was especially suitable for studying the kinetics of fermentation at low rates of sugar utilization. At high rates the fermentation system was unstable. 2. Glucose fermentation was fitted by a model in which a diffusion barrier with first-order kinetics is interposed between the environment and the site of hexokinase. 3. Aerobic conditions affected the fermentation enzyme system but not the diffusion mechanism. 4. The kinetics of maltose fermentation at low rates approximated to those of the hydrolysis of maltose by the enzyme maltase, as studied in suspensions of broken cells.

Colorimetry↗