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Continuous immobilized yeast reactor system for complete beer fermentation using spent grains and corncobs as carrier materials.

Despite extensive research carried out in the last few decades, continuous beer fermentation has not yet managed to outperform the traditional batch technology. An industrial breakthrough in favour of continuous brewing using immobilized yeast could be expected only on achievement of the following process characteristics: simple design, low investment costs, flexible operation, effective process control and good product quality. The application of cheap carrier materials of by-product origin could significantly lower the investment costs of continuous fermentation systems. This work deals with a complete continuous beer fermentation system consisting of a main fermentation reactor (gas-lift) and a maturation reactor (packed-bed) containing yeast immobilized on spent grains and corncobs, respectively. The suitability of cheap carrier materials for long-term continuous brewing was proved. It was found that by fine tuning of process parameters (residence time, aeration) it was possible to adjust the flavour profile of the final product. Consumers considered the continuously fermented beer to be of a regular quality. Analytical and sensorial profiles of both continuously and batch fermented beers were compared.

Beer↗

Immobilized yeast cell systems for continuous fermentation applications.

In several yeast-related industries, continuous fermentation systems offer important economical advantages in comparison with traditional systems. Fermentation rates are significantly improved, especially when continuous fermentation is combined with cell immobilization techniques to increase the yeast concentration in the fermentor. Hence the technique holds a great promise for the efficient production of fermented beverages, such as beer, wine and cider as well as bio-ethanol. However, there are some important pitfalls, and few industrial-scale continuous systems have been implemented. Here, we first review the various cell immobilization techniques and reactor setups. Then, the impact of immobilization on cell physiology and fermentation performance is discussed. In a last part, we focus on the practical use of continuous fermentation and cell immobilization systems for beer production.

Beer↗

Effects of colonic fermentation on respiratory gas exchanges following a glucose load in man.

Colonic fermentation produces short-chain fatty acids (SCFA). In humans, the amount of energy produced from the oxidation of these compounds is unknown and could modify the metabolic utilization of energetic fuels (eg, carbohydrates and lipids). If it were so, the equations used to evaluate the oxidation of nutrients from indirect calorimetry data should include the contribution of SCFA, which is not usually the case. Indeed, this fermentation process is usually considered as a minor and neglected energetic pathway. In this study, we have addressed the reliability of this assumption. Six normal subjects received orally either 50 g glucose or 50 g glucose plus 20 g lactulose. Their respiratory gas exchanges, breath hydrogen, methane, and 13CO2 concentrations, and plasma glucose, insulin, and free fatty acid (FFA) concentrations were monitored for 8 hours. CO2 production and breath hydrogen concentration were significantly greater with lactulose. No differences in oxygen consumption, breath 13CO2 production, or plasma concentrations of blood glucose, FFA, and insulin could be found between the two experiments. This suggests that the fermentation process induced by lactulose generates extra fuels going through an oxidation pathway. Therefore, the classic equations used to calculate carbohydrate and lipid oxidation and energy expenditure (EE) from indirect calorimetry data are probably not valid when fermentation is taking place. Indeed, in this experiment we could have overestimated glucose oxidation (12.5%) if the fermentation process were not considered. In conclusion, colonic fermentation in humans of nondigestible carbohydrates produces energetic substrates that could be used and oxidized as energetic fuels.(ABSTRACT TRUNCATED AT 250 WORDS)

Adolescent↗

Fermentation of cereals for reduction of bacterial contamination of weaning foods in Ghana.

Unfermented and fermented maize dough weaning foods prepared by mothers in a Ghanaian village were examined for gram-negative bacilli (GNB) immediately after preparation and during storage to assess the antimicrobial effect of fermentation. GNB were cultured from all samples of unfermented dough (51) and from 16 of 51 samples of fermented dough. The extent of contamination was significantly higher in the unfermented dough than in fermented dough (5.9 [SEM 0.1] vs 4.0 [0.4] log10 colony forming units/g). After 6 h and 12 h storage, a higher proportion of samples of porridge made from unfermented dough contained GNB than did those made with fermented dough (45/51 vs 22/55; 49/51 vs 20/51, respectively) and levels of GNB were significantly higher in the porridge made from unfermented dough after 6 h (4.2 [0.2] vs 3.8 [0.2]). Fermentation of maize dough is an effective method to reduce contamination of maize dough weaning foods with GNB.

Diarrhea, Infantile↗

Accelerated natural lactic fermentation of cereal-based formulas at reduced water activity.

Energy required to dehydrate fermented cereal-based food formulas plays a significant part in the production cost. Therefore the effect of reduced moisture content on the fermentation was investigated. Generally, lactic fermentation at reduced moisture content resulted in increased final pH. Significant acidification still occurred at 0.33 kg water/kg dry matter corresponding to either aW = 0.925 in a sorghum-maize-soya (SMS) mixture, or aW = 0.950 in a sorghum-maize-milk powder (SMM) mixture. Acidification adequate for microbiological safety (pH < or = 4.5) was achieved at 0.54 kg water/kg dm in SMS (aW = 0.950) and 0.43 kg/kg dm in SMM (aW = 0.965). In stable accelerated natural lactic fermentations obtained by inoculum recycling ('back-slopping') at 30 degrees C, dominant lactic acid bacteria included Lactobacillus plantarum, L. brevis, L. acidophilus and Lactococcus lactis. Dominating yeasts were Saccharomyces cerevisiae and Candida krusei. L. plantarum isolated from high-moisture (1.5 kg/kg dry matter) SMS fermentations had higher specific growth rates than L. plantarum isolated from reduced moisture (0.54 kg/kg dm) SMS fermentations, when tested under similar aW conditions. This effect was not caused by previous culturing conditions; probably different strains of L. plantarum having different aW-optima dominate at high or reduced moisture conditions.

Edible Grain↗

Significance of yeasts and moulds occurring in maize dough fermentation for 'kenkey' production.

Yeasts and moulds associated with the fermentation of maize dough during the processing of the West African traditional food 'kenkey' were investigated. A mixed flora comprising Candida, Saccharomyces, Trichosporon, Kluyveromyces and Debaryomyces species were isolated from raw maize, during steeping and early phases of fermentation. After 24-48 h of fermentation, Candida krusei and Saccharomyces cerevisiae dominated reaching counts exceeding 10(6) cfu/g. This succession of yeast populations and the significant multiplication of C. krusei and S. cerevisiae were observed in all cases for both the fermentation and the production sites investigated. Penicillium, Aspergillus and Fusarium species, including potential mycotoxin producers, were isolated from raw maize. Initial high counts of 10(5) cfu/g for moulds were reduced to less than 10(2)cfu/g within 24 h of fermentation. High levels of aflatoxins were observed in raw maize, and they were not affected during the fermentations.

Aflatoxins↗

Effect of fermentation conditions on the production of citric acid from cheese whey by Aspergillus niger.

The effect of pH value, methanol, and salt concentration on the production of citric acid from cheese whey by two strains of Aspergillus niger, i.e. CAIM 111 and CAIM 167, was investigated. Lactose concentration, utilized lactose, citric acid concentration, conversion coefficient of lactose to citric acid, and mycelial dry weight were measured during the fermentation process. The maximum citric acid concentration (1.06 and 0.82 g/l), and conversion coefficient (5.58 and 7.45%) were obtained at pH 3.5 after 9 days of fermentation for A. niger CAIM 111 and A. niger CAIM 167, respectively. The presence of 4% (v/v) methanol in the fermentation medium increased the amount of citric acid produced by A. niger CAIM 111 and A. niger CAIM 167 by 23% and 18%, respectively. Both strains showed a high ability to utilize lactose for the production of citric acid when grown in the presence of 10% (w/v) salt. The conversion coefficient of lactose to citric acid was 28.24% for A. niger CAIM 111 and 25.60% for A. niger CAIM 167 when the fermentation medium had a 10% (w/v) level of salt. The cumulative effect of fermentation medium pH (3.5), methanol concentration (4%, v/v) and salt concentration (10%, w/v) during the fermentation process of whey did not enhance the production of citric acid by A. niger CAIM 111, while it did increase the production of citric acid by A. niger CAIM 167 by about 4-fold.

Aspergillus niger↗

Ethanolic fermentation of acid pre-treated starch industry effluents by recombinant Saccharomyces cerevisiae strains.

Two industrial effluents, a pre-fermentation effluent and a post-fermentation effluent from a wheat starch production plant, were used as substrates for fuel ethanol production in anaerobic batch cultures using minimal nutritional amendment. The performances of three metabolically engineered xylose-utilizing Saccharomyces cerevisiae strains: TMB 3001 expressing XYL1, XYL2 and XKS1, redox metabolism modulated CPB.CR1 and glucose de-repressed CPB.CR2, as well as a reference strain CEN.PK 113-7D not fermenting xylose, were evaluated. For the recombinant strains a glucose consumption phase preceded the xylose consumption phase. In both effluents, biomass and ethanol production occurred predominantly during the glucose consumption phase, whereas xylitol and glycerol formation were predominant in the xylose consumption phase. Total specific ethanol productivities on glucose were 6-fold higher than on xylose in the pre-fermentation effluent and 15-fold higher than on xylose in the post-fermentation effluent. CPB.CR1 showed impaired growth compared to the two other xylose-utilizing strains, but displayed 18% increased ethanol yield in the post-fermentation effluent.

Biomass↗

Discrepancy in glucose and fructose utilisation during fermentation by Saccharomyces cerevisiae wine yeast strains.

While unfermented grape must contains approximately equal amounts of the two hexoses glucose and fructose, wine producers worldwide often have to contend with high residual fructose levels (>2 gl(-1)) that may account for undesirable sweetness in finished dry wine. Here, we investigate the fermentation kinetics of glucose and fructose and the influence of certain environmental parameters on hexose utilisation by wine yeast. Seventeen Saccharomyces cerevisiae strains, including commercial wine yeast strains, were evaluated in laboratory-scale wine fermentations using natural Colombard grape must that contained similar amounts of glucose and fructose (approximately 110 gl(-1) each). All strains showed preference for glucose, but to varying degrees. The discrepancy between glucose and fructose utilisation increased during the course of fermentation in a strain-dependent manner. We ranked the S. cerevisiae strains according to their rate of increase in GF discrepancy and we showed that this rate of increase is not correlated with the fermentation capacity of the strains. We also investigated the effect of ethanol and nitrogen addition on hexose utilisation during wine fermentation in both natural and synthetic grape must. Addition of ethanol had a stronger inhibitory effect on fructose than on glucose utilisation. Supplementation of must with assimilable nitrogen stimulated fructose utilisation more than glucose utilisation. These results show that the discrepancy between glucose and fructose utilisation during fermentation is not a fixed parameter but is dependent on the inherent properties of the yeast strain and on the external conditions.

Ethanol↗

Influence of yeast strain on ochratoxin A content during fermentation of white and red must.

The aim of this work was to examine whether the yeast strains, responsible for alcoholic fermentation, have an influence on the concentration of ochratoxin A (OTA) in wine. Before the fermentation, OTA was added to musts up to a concentration of about 2 microg/l. OTA content was determined in white and red wines resulting from respective musts and in methanolic extract of the yeast lees (MEL). Data showed a significant reduction of OTA at the end of alcoholic fermentation. However, depending on the yeast strain involved in the fermentation, there was a difference in the content of OTA in the wines. The percentage of OTA removal during the fermentation was between 46.83% and 52.16% in white wine and between 53.21% and 70.13% in red wine. The absence of degradation products suggested an adsorption mechanism. OTA concentration in MEL resulting from red must fermentation was higher than in white. A significant amount of OTA was not recovered either from wine or from MEL.

Adsorption↗

Changes in volatile compounds and related biochemical profile during controlled fermentation of cv. Conservolea green olives.

The effect of controlled fermentation processes on the profile of volatile and other biochemical compounds of cv. Conservolea green olives processed by the Spanish method was studied. The different treatments included: (a) inoculation with a commercial starter culture of Lactobacillus pentosus, (b) inoculation with a wild strain of Lactobacillus plantarum isolated from a previous fermentation, (c) uninoculated spontaneous process (control). Microbial growth, pH, titratable acidity, reducing sugars, organic acids and volatile compounds were monitored. Starter cultures were effective in establishing an accelerated fermentation process. Both were able to reduce the survival period of Enterobacteria by 7 days, minimizing thus the likelihood of spoilage. Higher acidification of the brines and faster pH drop was observed in inoculated processes, with L. pentosus presenting better performance than the wild strain of L. plantarum. Lactic and acetic were the major organic acids detected by HPLC, the concentration of which increased in the course of fermentation. Citric and malic acids were also present in the brines but they were degraded completely within the first 2 weeks of fermentation. Ethanol, methanol, acetaldehyde, ethyl acetate, isobutyric acid were the major volatile compounds identified by GC. Their concentration varied greatly among the fermentation processes, reflecting varying degrees of microbial activity in the brines.

Chromatography, High Pressure Liquid↗

Bioactive macromolecules in LAB-fermented cereals: Mechanisms of formation, functional properties, and health benefits.

Cereal and pseudo-cereal based fermented food products represent a substantial segment of global diet, nutrition as well as food security. Fermentation, especially by Lactic Acid Bacteria (LAB) increases the nutritional and functional values of foods by increasing palatability, bioavailability and minimizing antinutritional factors. LAB plays a pivotal role in synthesizing bioactive peptides, vitamins, minerals and reducing anti-nutrients parallelly. This review elucidates the mechanism through which LAB revamping nutritional macromolecules, such as peptides and polysaccharides, during fermentation and their role in the development of traditional as well as modern fermented foods. Additionally, these fermented foods have been associated with several health benefits. Recent advancement in biotechnology such as genome sequencing, functional genomics, and AI-assisted bioinformatics, have significantly enhanced our understanding of the diversity of LAB, the metabolism, and adaptation mechanisms. The combination of in silico and experimental methods has enabled the development of novel food enzymes as well as highly precise fermentation processes. Together with new innovations, growing demands for quality, consistency, safety as well as health benefits point out the significance of continued research. More studies employing both conventional and modern methods are necessary to explore these food groups completely and achieve better food quality, increased nutrition, more health benefits and comprehensive socioeconomic advantages.

Bioactive macromolecules↗

Effect of boiling and roasting on the fermentation of soybeans into dawadawa (soy-dawadawa).

Soybeans which had initially been dehulled by either boiling (boiled/dehulled) or roasting (roasted/dehulled) before peeling, were cooked and fermented into dawadawa, a traditional food condiment. The micropopulation, enzymatic activities, proximate composition, amino acid, and aroma profiles of the two types of soybean dawadawa were evaluated during fermentation. Only minor differences were found in the microbial profiles of the two types of soy-dawadawa. Although boiled/dehulled soy-dawadawa initially had lower microbial counts, it recorded higher counts at the advanced stages of fermentation. Proteolytic and amylolytic Bacillus species including Bacillus subtilis, Bacillus pumilus, Bacillus licheniformis, Bacillus cereus, and Bacillus firmus dominated the micropopulation of the two types of soy-dawadawa with Bacillus subtilis accounting for about 50% of the Bacillus species in all samples. Lactic acid bacteria and yeasts occurred in low numbers in the two types of soy-dawadawa. The proximate composition of the two types of soy-dawadawa were similar, and their contents of moisture and protein increased whilst fat and ash decreased during fermentation. Both types of fermenting soy-dawadawa recorded similar levels of alpha-amylase activity, but boiled/dehulled soy-dawadawa showed slightly higher protease activity. The levels of isoleucine, leucine, lysine, phenylalanine, arginine and proline increased significantly with fermentation time in both types of soy-dawadawa. With respect to differences in their aroma profiles, hexanodecanol, octadecyl acetate, 1,2-dimethyl benzene, tetradecene, (E)-5-eicosene, cyclohexadecane, and hexacosane were found only in the roasted/dehulled samples, whilst 1,2-ethanediol, ethyl acetate, dimethyl disulfide, cyclotetradecane, decene, indole , 2 butyl-octenal, acetophenone, and toluene were found only in the boiled/dehulled samples. A market focus group showed preference for roasted/dehulled soy-dawadawa over boiled/dehulled soy-dawadawa. Apart from the volatile aroma compounds, the biochemical and microbiological profiles of the two types of soy-dawadawa showed only minor differences and were also similar to the profiles reported for African locust bean dawadawa.

Amino Acids↗

Identification of potential probiotic starter cultures for Scandinavian-type fermented sausages.

Potential probiotic cultures suitable as starter cultures for the Scandinavian-type fermented sausages were identified among strains well-adapted to fermented meats as well as strains originating from a culture collection. From 15 different fermented meat products, 22 strains were isolated as dominant non-starter lactic acid bacteria (NSLAB). The isolates were identified by RAPD, API and sequence analysis of 16S rRNA and showed to be five strains of Lactobacillus sakei, five strains of Lactobacillus farciminis, five strains belonging to the group of Lactobacillus plantarum/pentosus, four strains of Lactobacillus alimentarius, two strains of Lactobacillus brevis and one strain of Lactobacillus versmoldensis. Heterofermentative strains as well as strains not growing at 37 degrees C and not lowering pH below 5.1 in a meat model were excluded leaving 9 strains for further studies. These strains together with 19 strains from a culture collection were evaluated by in vitro methods including survival upon exposure to pH 2.5 or 0.3% oxgall and adhesion to the human colon adenocarcinoma cell line Caco-2 as well as antimicrobial activity against potential pathogens. Strains that fulfilled all the probiotic criteria and showed to be fast acid producers in a meat model included three strains belonging to the group of Lb. plantarum/pentosus (MF1291, MF1298, MF1300) which originated from the dominant NSLAB of fermented meat products. MF1291 and MF 1298 were further identified as Lb. plantarum and MF1300 as Lb. pentosus. The three strains were all successfully applied as starter cultures for the production of fermented sausage. The viable count at the end of the processing period reached high cell numbers (4.7x10(7)-2.9x10(8) cfu/g) and pH of the sausages decreased to pH 4.8-4.9 without any flavour deviation compared to sausage fermented by a commercial meat starter culture.

Animals↗

Technological characterization of a bacteriocin-producing Lactobacillus sakei and its use in fermented sausages production.

The aim of this paper was the technological characterization of a Lactobacillus sakei strain, able to produce the bacteriocin sakacin P, that was originally isolated from naturally fermented sausages. Experiments were conducted in situ, using MRS-based medium, and in situ, when the strain was inoculated as starter culture in real sausage fermentation. The results obtained underlined that the strain was able to grow in conditions that are commonly used in the production line, and only lactose and high concentrations of NaCl (5% w/v) reduced the capability for bacteriocin production. When inoculated in sausages, the strain showed a good performance, being able to colonize rapidly the ecosystem. A high number of isolates, capable of producing sakacin P, were already isolated after the third day of fermentation, and persisted throughout the course of the fermentation. The inoculated strain also affected other microbial colonization trends; in fact the total bacterial count and fecal enterococci showed a rapid decrease at the end of the fermentation. Moreover, during sensory evaluation, the final sausage product received high scores for the parameters of tenderness and juiciness, with medium acidity and low rancidity. Lastly, the panelists preferred the sausages produced with the L. sakei characterized in this study when compared to a fermented sausage produced with a commercial starter.

Animals↗

Study on the microflora and biochemistry of cocoa fermentation in the Dominican Republic.

Cocoa fermentation was monitored at the IDIAF (Instituto Dominicano de Investigaciones Agropecuarias y Forestales) "Mata Larga" experimental station, in San Francisco de Macoris, Dominican Republic. The maximum average fermentation temperature reached 51 degrees C after 48 h and the pH reached 4.5 after 144 h of fermentation. A significant decrease in glucose, fructose and citric acid was seen in the pulp over the first 48 h. There was a delay of 24 h between maximum microbial growth and maximum concentrations of the respective metabolites, which occurred after 48 h for ethanol and after 72 h for acetic acid. A maximum concentration in lactic acid was found after around 120 h of fermentation. The aerobic mesophilic flora increased from 6.1x10(6) to a maximum of 4.2x10(7) CFU g(-1) of dry matter after 48 h of fermentation. Yeasts displayed maximum development after 24 h (6.1x10(7) CFU g(-1) of dry matter), whilst for lactic and acetic acid bacteria it occurred after 48 h (7.3x10(7) and 1.5x10(8) CFU g(-1) of dry matter respectively). The yeasts isolated belonged to the genera Hanseniaspora and Candida, the lactic acid bacteria to the genus Lactobacillus, and the acetic acid bacteria to the genus Acetobacter. The differences compared to other fermentation trials concerned the micropopulation from a qualitative point of view.

Bacteria, Aerobic↗

Survival of Escherichia coli O157:H7 during the fermentation of Spanish-style green table olives (conservolea variety) supplemented with different carbon sources.

Olives were supplemented with two different carbon sources (sucrose and glucose) at five different levels (0%, 0.1%, 0.3%, 0.5% and 1%) and the fermentation procedure was followed after inoculation with or without starter culture. Inhibition of Escherichia coli O157:H7 was evident in all fermentation procedures regardless of the treatment used. The pathogen numbers declined but did not die out during fermentation. It was evident that the rate of death was higher in samples supplemented with starter cultures compared to natural fermentation. The production of acids during fermentation seems to be the main factor that governs the behaviour of this pathogen under such stress conditions. The HPLC analysis revealed that lactate, formate and acetate were among the end-products during the fermentation of green olives.

Acetates↗

Appropriate starter culture technologies for small-scale fermentation in developing countries.

Modern food biotechnology has moved a long way since ancient times of empirical food fermentations. Preservation and safeguarding of food are, however, still major objectives of fermentation. In addition, other aspects, such as wholesomeness, acceptability and overall quality, have become increasingly important and valued features to consumers even in developing countries where old traditions and cultural particularities in food fermentations are generally well maintained. Due to limitations in infrastructure and existing low technologies, rural areas in most developing countries have not been able to keep abreast of global developments toward industrialisation. At the same time, fermented foods play a major role in the diet of numerous regions in Africa and Asia. In many traditional approaches, the advantages of some form of inoculation of a new batch, e.g. by back-slopping or the repeated use of the same container (e.g. a calabash) is appreciated and generally practised. Still, the benefits of small-scale starter culture application as a means of improved hygiene, safety and quality control, in support of HACCP approaches, are not yet realised in small-scale fermentation operations. Approaches and considerations for the selection of pure cultures for small-scale, low-tech applications may differ in some respects from the large-scale industrial approaches practised since 100 years. Selection criteria should take account of the substrate, technical properties of the strain, food safety requirements and quality expectations. Lack of experience in the application of starter cultures in small-scale operations and under rural conditions presents a major obstacle but also an exciting challenge to food microbiologist and technologist. Culture preservation, maintenance and distribution demand special logistic and economic considerations. Quality, safety and acceptability of traditional fermented foods may be significantly improved through the use of starter cultures selected on the basis of multifunctional considerations, also taking into account the probiotic concept and possibilities offered for improved health benefits.

Bacteria↗