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PubMed2003

Antimicrobial interactions of microbial species involved in the fermentation of cassava dough into agbelima with particular reference to the inhibitory effect of lactic acid bacteria on enteric pathogens.

Lactic acid bacteria, Bacillus species and yeasts are involved in the fermentation of cassava dough into agbelima. Microbial interactions within and between these groups of microorganisms were investigated in addition to the survival of five enteric pathogens inoculated into agbelima under various conditions. Nine out of 10 cultures of lactic acid bacteria isolated at the end of agbelima fermentation showed inhibitory effect against 10 cultures of lactic acid bacteria isolated at the start of fermentation. Only 3 out of 10 isolates of Bacillus subtilis were inhibited by 10 isolates of lactic acid bacteria tested. No interactions were observed between yeasts and the lactic acid bacteria, whereas three of the Bacillus isolates showed inhibitory effects against the yeasts. Twelve isolates of Lactobacillus plantarum tested inhibited the growth of an isolate each of Lactobacillus fermentum and Lactobacillus brevis but none tested positive for bacteriocin production. The antimicrobial effect of the lactic acid bacteria was attributed to acid production. In fermenting cassava dough, enteric pathogens survived to different extents depending on pH and their sensitivity to acids. Vibrio cholerae C-230, Salmonella typhimurium 9 and Salmonella enteritidis 226 were not detectable in 10 g of sample after 4 h when inoculated into the 48-h fermented product, agbelima, whereas Shigella dysenteriae 2357T and Escherichia coli D2188 were detectable up to 8 h in the product.

Antibiosis↗
PubMed2003

Microbial modification of the texture of grated cassava during fermentation into akyeke.

The traditional akyeke inoculum and fermenting akyeke, an indigenous cassava product, were investigated to identify microbial species responsible for the modification of cassava texture during fermentation. Both field and laboratory samples were examined and only some cultures isolated on Plate Count Agar and Malt Extract Agar were found to be capable of causing a softening of cassava tissue when plated directly on sterile cassava slices. The cassava tissue softening isolates on PCA were tentatively identified as Bacillus subtilis and isolates on MEA as Candida tropicalis and Zygosacchromyces florentinus. The population of B. subtilis in the laboratory sample of inoculum was found to be 2.4 x 10(9) cfu g(-1) and increased during dough fermentation from 1.1 x 10(7) to 3.5 x 10(9) cfu g(-1) after 96 h. C. tropicalis was present in the inoclum at 3.0 x 10(9) cfu g(-1) and increased during dough fermentation from 3.2 x 10(6) to 6.9 x 10(7) cfu g(-1) whilst Z. florentinus was present in the inoclum at 9.1 x 10(8) cfu g(-1) and increased from 8.1 x 10(5) to 7.5 x 10(6) cfu g(-1) during dough fermentation.

Bacillus subtilis↗
PubMed1998

Lactic acid fermentation and storage of blanched garlic.

The controlled fermentation of peeled, blanched garlic, using a starter culture of Lactobacillus plantarum, was studied and compared with that of unblanched garlic. Blanching was carried out in hot water (90 degrees C) for 15 min. The starter grew abundantly in the case of blanched garlic, producing mainly lactic acid and reaching a pH of 3.8 after 7 days, but its growth was inhibited in unblanched garlic. Ethanol and fructose, coming from enzymatic activities of the garlic, and a green pigment were formed during the fermentation of unblanched garlic, but not of blanched garlic. The blanched garlic fermented by L. plantarum, even without a preservation treatment (pasteurization), was microbiologically stable during storage at 30 degrees C in an acidified brine (approximately 3% (w/w) NaCl and pH 3.5 at equilibrium), but fructans were hydrolyzed. The packed fermented product and that obtained by direct packing without fermentation were not significantly different with regard to flavour.

Carbohydrates↗
PubMed1999

Chromosomal structures of bottom fermenting yeasts.

A genomic comparison of bottom fermenting yeasts was performed by pulsed-field gel electrophoresis and Southern blot analysis with some S. cerevisiae gene probes. We confirmed that strains of bottom fermenting yeast have four chromosomes originating from S. bayanus. Since the structures of these chromosomes were recombined with S. cerevisiae chromosomes, these S. bayanus chromosomes could be differentiated from S. cerevisiae chromosomes using Southern hybridization. Our Southern hybridization results indicate that bottom fermenting yeasts have both chromosomes originating from both S. cerevisiae and S. bayanus. It was reconfirmed that top fermenting yeast should be classified as S. cerevisiae, based on the chromosomal structure. The chromosomal structure of S. pastorianus CBS1538, the type stain of S. pastorianus, was also investigated. This strain has chromosomes originating only from S. bayanus. S. carlsbergensis CBS1513 has chromosomes originating from both S. cerevisiae and S. bayanus. From these results, we contend that bottom fermenting yeasts should be classified as S. carlsbergensis.

Blotting, Southern↗
PubMed2002

High-temperature alcoholic fermentation of whey using Kluyveromyces marxianus IMB3 yeast immobilized on delignified cellulosic material.

A novel system for high-temperature alcoholic fermentation of whey is described. This system consists of Kluyveromyces marxianus yeast immobilized on delignified cellulosic material (DCM). The effect of pH, initial lactose concentration and temperature on the fermentation of a synthetic medium containing lactose was studied. Batch fermentations of whey were also carried out and the formation of volatile by-products was examined. The concentrations of higher alcohols were found to be in very low levels leading to a product of improved quality. The fermented whey had an improved characteristic aroma compared to unfermented whey. The possibility to use fermented whey as raw material for the production of a novel, low alcohol content drink was also investigated.

Alcohols↗
PubMed2003

Replacement of maize by rumen filtrate fermented corn-cob in layer diets.

Laying hens (50 weeks in-lay) were fed for 8 weeks on diets containing corn-cobs fermented with rumen filtrate. The fermented corn-cob was included in the diets at 0%, 5.0%, 10.0%, 15.0% and 20.0% of the total diet at the expense of maize. Feed intake was not significantly (P>0.05) different, although birds on the control diet consumed slightly more feed than birds on the fermented corn-cob based diets. Hen day production and feed per dozen eggs decreased with increase in dietary level of corn-cob (P<0.05). Yolk colour was significantly (P<0.05) affected, rumen filtrate fermented corn-cob appear to promote better yolk coloration. Egg weight and shell thickness were not significantly influenced by dietary treatment. The overall results appear to suggest that rumen filtrate fermentated corn-cob could provide a material that holds promise as a good alternative to maize because of its enhanced crude protein value and reduced crude fibre content.

Animal Feed↗
PubMed2003

Occurrence and taxonomic characteristics of strains of Saccharomyces cerevisiae predominant in African indigenous fermented foods and beverages.

Indigenous fermented foods and beverages play a major role in the diet of African people. The predominant yeast species seen is Saccharomyces cerevisiae, involved in basically three groups of indigenous fermented products: non-alcoholic starchy foods, alcoholic beverages and fermented milk. These products are to a great extent made by spontaneous fermentation and consequently S. cerevisiae often coexists with other microorganisms even though a microbiological succession usually takes place both between and within species. The functions of S. cerevisiae are mainly related to formation of alcohols and other aroma compounds, but stimulation of e.g. lactic acid bacteria, improvement of nutritional value, probiotic effects, inhibition of undesired microorganisms and production of tissue-degrading enzymes may also be observed. Several different isolates of S. cerevisiae have been shown to be involved in the fermentations and some of the isolates show pheno- and genotypic characteristics that deviate from those normally recognised for S. cerevisiae.

Africa South of the Sahara↗
PubMed2003

The impact of brewing yeast cell age on fermentation performance, attenuation and flocculation.

Individual cells of the yeast Saccharomyces cerevisiae exhibit a finite replicative lifespan, which is widely believed to be a function of the number of divisions undertaken. As a consequence of ageing, yeast cells undergo constant modifications in terms of physiology, morphology and gene expression. Such characteristics play an important role in the performance of yeast during alcoholic beverage production, influencing sugar uptake, alcohol and flavour production and also the flocculation properties of the yeast strain. However, although yeast fermentation performance is strongly influenced by the condition of the yeast culture employed, until recently cell age has not been considered to be important to the process. In order to ascertain the effect of replicative cell age on fermentation performance, age synchronised populations of a lager strain were prepared using sedimentation through sucrose gradients. Each age fraction was analysed for the ability to utilise fermentable sugars and the capacity to flocculate. In addition cell wall properties associated with flocculation were determined for cells within each age fraction. Aged cells were observed to ferment more efficiently and at a higher rate than mixed aged or virgin cell cultures. Additionally, the flocculation potential and cell surface hydrophobicity of cells was observed to increase in conjunction with cell age. The mechanism of ageing and senescence in brewing yeast is a complex process, however here we demonstrate the impact of yeast cell ageing on fermentation performance.

Alcian Blue↗
PubMed2003

Xylose and cellobiose fermentation to ethanol by the thermotolerant methylotrophic yeast Hansenula polymorpha.

Wild-type strains of the thermotolerant methylotrophic yeast Hansenula polymorpha are able to ferment glucose, cellobiose and xylose to ethanol. H. polymorpha most actively fermented sugars to ethanol at 37 degrees C, whereas the well-known xylose-fermenting yeast Pichia stipitis could not effectively ferment carbon substrates at this temperature. H. polymorpha even could ferment both glucose and xylose up to 45 degrees C. This species appeared to be more ethanol tolerant than P. stipitis but more susceptible than Saccharomyces cerevisiae. A riboflavin-deficient mutant of H. polymorpha increased its ethanol productivity from glucose and xylose under suboptimal supply with riboflavin. Mutants of H. polymorpha defective in alcohol dehydrogenase activity produced lower amounts of ethanol from glucose, whereas levels of ethanol production from xylose were identical for the wild-type strain and the alcohol dehydrogenase-defective mutant.

Cellobiose↗
PubMed2003

New quinoproteins in oxidative fermentation.

Several quinoproteins have been newly indicated in acetic acid bacteria, all of which can be applied to fermentative or enzymatic production of useful materials by means of oxidative fermentation. (1) D-Arabitol dehydrogenase from Gluconobacter suboxydans IFO 3257 was purified from the bacterial membrane and found to be a versatile enzyme for oxidation of various substrates to the corresponding oxidation products. It is worthy of notice that the enzyme catalyzes D-gluconate oxidation to 5-keto-D-gluconate, whereas 2-keto-D-gluconate is produced by a flavoprotein D-gluconate dehydrogenase. (2) Membrane-bound cyclic alcohol dehydrogenase was solubilized and purified for the first time from Gluconobacter frateurii CHM 9. When compared with the cytosolic NAD-dependent cyclic alcohol dehydrogenase crystallized from the same strain, the reaction rate in cyclic alcohol oxidation by the membrane enzyme was 100 times stronger than the cytosolic NAD-dependent enzyme. The NAD-dependent enzyme makes no contribution to cyclic alcohol oxidation but contributes to the reduction of cyclic ketones to cyclic alcohols. (3) Meso-erythritol dehydrogenase has been purified from the membrane fraction of G. frateurii CHM 43. The typical properties of quinoproteins were indicated in many respects with the enzyme. It was found that the enzyme, growing cells and also the resting cells of the organism are very effective in producing L-erythrulose. Dihydroxyacetone can be replaced by L-erythrulose for cosmetics for those who are sensitive to dihydroxyacetone. (4) Two different membrane-bound D-sorbitol dehydrogenases were indicated in acetic acid bacteria. One enzyme contributing to L-sorbose production has been identified to be a quinoprotein, while another FAD-containing D-sorbitol dehydrogenase catalyzes D-sorbitol oxidation to D-fructose. D-Fructose production by the oxidative fermentation would be possible by the latter enzyme and it is superior to the well-established D-glucose isomerase, because the oxidative fermentation catalyzes irreversible one-way oxidation of D-sorbitol to D-fructose without any reaction equilibrium, unlike D-glucose isomerase. (5) Quinate dehydrogenase was found in several Gluconobacter strains and other aerobic bacteria like Pseudomonas and Acinetobacter strains. It has become possible to produce dehydroquinate, dehydroshikimate, and shikimate by oxidative fermentation. Quinate dehydrogenase was readily solubilized from the membrane fraction by alkylglucoside in the presence of 0.1 M KCl. A simple purification by hydrophobic chromatography gave a highly purified quinate dehydrogenase that was monodispersed and showed sufficient purity. When quinate dehydrogenase purification was done with Acinetobacter calcoaceticus AC3, which is unable to synthesize PQQ, purified inactive apo-quinate dehydrogenase appeared to be a dimer and it was converted to the monomeric active holo-quinate dehydrogenase by the addition of PQQ.

Alcohol Oxidoreductases↗
PubMed2000

In vitro fermentability of dextran, oligodextran and maltodextrin by human gut bacteria.

Anaerobic batch culture fermenters were used for a preliminary screening of the in vitro utilization by human gut microflora of dextran and novel oligodextrans (I, II and III) produced in the University of Reading (UK). Glucose and fructooligosaccharides (FOS) were used as reference carbohydrates. As expected, FOS acted as a good prebiotic in that it selectively increased numbers of bifidobacteria in the early stages of the fermentation. Dextran and oligodextrans each resulted in an enrichment of bifidobacteria in the batch cultures, with high levels of persistence up to 48 h. They also produced elevated levels of butyrate ranging from 5 to 14.85 mmol/l. To more effectively simulate conditions that prevail in different regions of the large intestine, a three-stage continuous culture cascade system was used to study further the fermentation of dextran, a low-molecular-mass oligodextran (IV) and maltodextrin. Oligodextran IV was shown to be the best substrate for bifidobacteria and lactobacilli with steady-state populations of bifidobacteria and lactobacilli being higher in all three vessels of the gut model than the respective populations resulting from dextran and maltodextrin. A maximum difference of 1.9 log was observed in vessel 1 for both bifidobacteria and lactobacilli in the case of dextran fermentation, with 1.4 log and 0.8 log in vessel 3 were the maximum differences for bifidobacteria and lactobacilli when maltodextrin was used as the carbohydrate source. Moreover, dextran and oligodextran appeared to stimulate butyrate production, with a maximum production up to 25.39 mmol/l in vessel 3 when fermenting dextran, followed by 21.70 mmol/l in the case of oligodextran IV and only 12.64 mmol/l in the case of maltodextrin.

Bifidobacterium↗
PubMedDate not supplied

Effects of glucose, vitamins, and DO concentrations on pyruvate fermentation using Torulopsis glabrata IFO 0005 with metabolic flux analysis.

The effects of glucose, vitamins, and DO concentrations on efficient pyruvic acid fermentation were investigated using Torulopsis glabrata IFO 0005, and a novel biphasic culture method was developed on the basis of the metabolic flux analysis. T. glabrata requires the four vitamins nicotinic acid (NA), thiamine hydrochloride (B(1)), pyridoxine hydrochloride, and biotin for cell growth. The deficiency of these vitamins plays an essential role in pyruvate fermentation. In the present study, we considered the effects of the first two vitamins on the pyruvate fermentation. On the basis of several batch and fed-batch experiments, it was found that, as a result of glucose inhibition of cell growth, the initial glucose concentration should be around 30-40 g/L, and the glucose concentration during fermentation should be controlled at high level around 30 g/L. On the basis of an analysis of carbon flux distribution, a biphasic fermentation method was developed where the cultivation started with a high DO (at 40-50% of air saturation) for efficient cell growth and then was reduced to 5-10% for efficient pyruvate production. Since a fair amount of ethanol was formed when the DO concentration was decreased, the addition of NA turned out to be effective in reducing the ethanol formation. This may be due to a relaxing of the requirement for NADH oxidation by the alcohol dehydrogenase pathway. Since B(1) affects both the pyruvate dehydrogenase complex and pyruvate decarboxylase, its initial concentration must be carefully determined by considering both the cell growth and pyruvate production phases.

Candida↗
PubMedDate not supplied

Metabolic flux variation of Saccharomyces cerevisiae cultivated in a multistage continuous stirred tank reactor fermentation environment.

The technique of metabolic flux analysis was implemented to elucidate the flux balancing of Saccharomyces cerevisiae cultivated in a multistage continuous stirred tank reactor fermentation environment. The results showed that the majority of the substrate (97.70 +/- 0.49%) was funneled into the glycolytic pathway, while the remainder was subdivided between the pentose phosphate pathway and pathways for polysaccharide synthesis. At the pyruvate node, 87.30 +/- 1.38% of the flux was channeled through the reaction governed by pyruvate decarboxylase. Fluxes through the pyruvate dehydrogenase bypass were maintained at a constant level (82.65 +/- 1.47%) irrespective of the configuration of the fermentation setup. Activity through the TCA "cycle" was replenished by the reaction catalyzed by pyruvate carboxylase and by the transport of cytosolic oxaloacetate across the mitochondrial membrane. The CO(2) evolution rate varied as fermentation progressed; however, the yield coefficient of CO(2) remained at a constant value. Although a constant yield of ethanol (0.42 g of ethanol/g of glucose) was obtained, operations of the TCA cycle were gradually switched from partially reductive to partially oxidative pathways from the first fermenter to the fourth fermenter.

Alcoholic Beverages↗
PubMedDate not supplied

Effects of increased impeller power in a production-scale Aspergillus oryzae fermentation.

The goal in this study was to determine how increased impeller power affects enzyme expression in large-scale (80 m(3)), fed-batch Aspergillus oryzae fermentations. An approximate 50% increase in average impeller power was achieved by increasing impeller diameter approximately 10%, while operating at slightly reduced speed. Measured decreases in terminal (95%) mixing time show increased power improved bulk mixing. However, batches operated at increased power had lower recombinant enzyme productivity. Biomass assays and image analysis tests showed no significant difference between "high power" and control batches, suggesting that slower growth, altered morphology, or increased hyphal fragmentation were not the cause of reduced productivity. Off-line tests on the shear-thinning, highly viscous broth show oxygen limitation occurred after transport through the air-liquid interface and imply the limitation may involve bulk mixing. Specifically, oxygen transfer may be limited to a small zone surrounding each impeller. When this is the case, oxygen mass transfer will be determined by both impeller shear and fluid circulation, which have been characterized with the energy dissipation/circulation function (EDCF). EDCF values during control fermentations were approximately constant at 25 kW m (-3) s(-1), while EDCF values during "high power" batches fell linearly from 40 to 15 kW m (-3) s(-1). The point at which "high power" EDCF values drop below those in control fermentations corresponds almost exactly with the point at which product titer stops increasing. Thus, our findings suggest oxygen mass transfer was less efficient during the latter half of "high power" fermentations because of reductions in impeller speed and subsequent decreases in EDCF values. This observation has clear implications during the scale-up of viscous fungal fermentations, implying that not only is the level of impeller power important, but also relevant is how this power is applied.

Aspergillus oryzae↗
PubMedDate not supplied

Production of phytase by Mucor racemosus in solid-state fermentation.

Phytase production was studied by three Mucor and eight Rhizopus strains by solid-state fermentation (SSF) on three commonly used natural feed ingredients (canola meal, coconut oil cake, wheat bran). Mucor racemosus NRRL 1994 (ATCC 46129) gave the highest yield (14.5 IU/g dry matter phytase activity) on coconut oil cake. Optimizing the supplementation of coconut oil cake with glucose, casein and (NH(4))(2)SO(4), phytase production in solid-state fermentation was increased to 26 IU/g dry matter (DM). Optimization was carried out by Plackett-Burman and central composite experimental designs. Using the optimized medium phytase, alpha-amylase and lipase production of Mucor racemosus NRRL 1994 was compared in solid-state fermentation and in shake flask (SF) fermentation. SSF yielded higher phytase activity than did SF based on mass of initial substrate. Because this particular isolate is a food-grade fungus that has been used for sufu fermentation in China, the whole SSF material (crude enzyme, in situ enzyme) may be used directly in animal feed rations with enhanced cost efficiency.

6-Phytase↗
PubMedDate not supplied

Assessment of in-line near-infrared spectroscopy for continuous monitoring of fermentation processes.

The application of NIR in-line to monitor and control fermentation processes was investigated. Determination of biomass, glucose, and lactic and acetic acids during fermentations of Staphylococcus xylosus ES13 was performed by an interactance fiber optic probe immersed into the culture broth and connected to a NIR instrument. Partial least squares regression (PLSR) calibration models of second derivative NIR spectra in the 700-1800 nm region gave satisfactory predictive models for all parameters of interest: biomass, glucose, and lactic and acetic acids. Batch, repeated batch, and continuous fermentations were monitored and automatically controlled by interfacing the NIR to the bioreactor control unit. The high frequency of data collection permitted an accurate study of the kinetics, supplying lots of data that describe the cultural broth composition and strengthen statistical analysis. Comparison of spectra collected throughout fermentation runs of S. xylosus ES13, Lactobacillus fermentum ES15, and Streptococcus thermophylus ES17 demonstrated the successful extension of a unique calibration model, developed for S. xylosus ES13, to other strains that were differently shaped but growing in the same medium and fermentation conditions. NIR in-line was so versatile as to measure several biochemical parameters of different bacteria by means of slightly adapted models, avoiding a separate calibration for each strain.

Acetic Acid↗
PubMedDate not supplied

Bacterial cellulose production by fed-batch fermentation in molasses medium.

Batch and fed-batch fermentations for bacterial cellulose (BC) production using molasses as a carbon source by Acetobacter xylinum BPR2001 were carried out in a jar fermentor. For improvement of BC production, molasses was subjected to H2SO4-heat treatment. The maximum BC concentration by this treated molasses increased 76%, and the specific growth rate increased 2-fold compared with that by untreated molasses. In batch fermentation, when the initial sugar concentrations of H2SO4-heat-treated molasses were varied from 20 to 70 g/L, the highest value of maximum BC concentration of 5.3 g/L was observed at 20 g/L. BC production in intermittent fed-batch (IFB) fermentation was conducted referring to the data in batch fermentation, and the highest BC production of 7.82 g/L was obtained when 0.2 L of molasses medium was added five times. When continuous fed-batch (CFB) fermentations were conducted, maximum BC concentration was obtained with a feeding rate of 6.3 g-sugar/h, which was derived from the optimal IFB experiment.

Bioreactors↗
PubMedDate not supplied

Continuous fermentation of undetoxified dilute acid lignocellulose hydrolysate by Saccharomyces cerevisiae ATCC 96581 using cell recirculation.

Saccharomyces cerevisiae ATCC 96581 was cultivated in a chemostat reactor with undetoxified dilute acid softwood hydrolysate as the only carbon and energy source. The effects of nutrient addition, dilution rate, cell recirculation, and microaerobicity were investigated. Fermentation of unsupplemented dilute acid lignocellulose hydrolysate at D = 0.10 h(-1) in an anaerobic continuous reactor led to washout. Addition of ammonium sulfate or yeast extract was insufficient for obtaining steady state. In contrast, dilute acid lignocellulose hydrolysate supplemented with complete mineral medium, except for the carbon and energy source, was fermentable under anaerobic steady-state conditions at dilution rates up to 0.14 h(-1). Under these conditions, washout occurred at D = 0.15 h(-1). This was preceded by a drop in fermentative capacity and a very high specific ethanol production rate. Growth at all different dilution rates tested resulted in residual sugar in the chemostat. Cell recirculation (90%), achieved by cross-flow filtration, increased the sugar conversion rate from 92% to 99% at D = 0.10 h(-1). Nutrient addition clearly improved the long-term ethanol productivity in the recirculation cultures. Application of microaerobic conditions on the nutrient-supplemented recirculation cultures resulted in a higher production of biomass, a higher cellular protein content, and improved fermentative capacity, which further improves the robustness of fermentation of undetoxified lignocellulose hydrolysate.

Acetic Acid↗