Batch Culture Fermentation Research Articles

Continuous Butanol Fermentation of Dilute Acid-Pretreated De-oiled Rice Bran by Clostridium acetobutylicum YM1

Continuous fermentation of dilute acid-pretreated de-oiled rice bran (DRB) to butanol by the Clostridium acetobutylicum YM1 strain was investigated. Pretreatment of DRB with dilute sulfuric acid (1%) resulted in the production of 42.12 g/L total sugars, including 25.57 g/L glucose, 15.1 g/L xylose and 1.46 g/L cellobiose. Pretreated-DRB (SADRB) was used as a fermentation medium at various dilution rates, and a dilution rate of 0.02 h−1 was optimal for solvent production, in which 11.18 g/L of total solvent was produced (acetone 4.37 g/L, butanol 5.89 g/L and ethanol 0.92 g/L). Detoxification of SADRB with activated charcoal resulted in the high removal of fermentation inhibitory compounds. Fermentation of detoxified-SADRB in continuous fermentation with a dilution rate of 0.02 h−1 achieved higher concentrations of solvent (12.42 g/L) and butanol (6.87 g/L), respectively, with a solvent productivity of 0.248 g/L.h. This study showed that the solvent concentration and productivity in continuous fermentation from SADRB was higher than that obtained from batch culture fermentation. This study also provides an economic assessment for butanol production in continuous fermentation process from DRB to validate the commercial viability of this process.

Introducing a thermotolerant Gluconobacter japonicus strain, potentially useful for coenzyme Q10 production

In this report, Gluconobacter strains were screened for coenzyme Q10 (CoQ10) production. A thermotolerant strain, Gluconobacter japonicus FM10, was eventually employed for CoQ10 production optimization. To do so, a two-step optimization strategy was used. The first step focused on biomass increase and the second step focused on increase in CoQ10 production. Factors including temperature, pH, carbon, and nitrogen sources were optimized at the first step, and temperature, pH, and aeration were optimized at the second step. The batch culture fermentation was used with the optimized factors of the first phase (30°C, pH6.5, D-sorbitol, and yeast extract-peptone as the carbon and nitrogen sources). After 18h, the temperature, pH, and aeration were shifted to the optimized values of the second step (36°C, pH7, and no aeration). By this strategy, the dry cell mass (17.1g/L) and CoQ10 (23.2mg/L) were obtained after 20h, which the latter was 2.3 times higher than that of the first step of optimization. Among the conditions tested, carbon source was the most important factor on the cell growth at the first step while no aeration was the key factor for CoQ10 production in the second step of optimization.

Wood-Derived Dietary Fibers Promote Beneficial Human Gut Microbiota.

Woody biomass is a sustainable and virtually unlimited source of hemicellulosic polysaccharides. The predominant hemicelluloses in softwood and hardwood are galactoglucomannan (GGM) and arabinoglucuronoxylan (AGX), respectively. Based on the structure similarity with common dietary fibers, GGM and AGX may be postulated to have prebiotic properties, conferring a health benefit on the host through specific modulation of the gut microbiota. In this study, we evaluated the prebiotic potential of acetylated GGM (AcGGM) and highly acetylated AGX (AcAGX) obtained from Norwegian lignocellulosic feedstocks in vitro In pure culture, both substrates selectively promoted the growth of Bifidobacterium, Lactobacillus, and Bacteroides species in a manner consistent with the presence of genetic loci for the utilization of β-manno-oligosaccharides/β-mannans and xylo-oligosaccharides/xylans. The prebiotic potential of AcGGM and AcAGX was further assessed in a pH-controlled batch culture fermentation system inoculated with healthy adult human feces. Results were compared with those obtained with a commercial fructo-oligosaccharide (FOS) mixture. Similarly to FOS, both substrates significantly increased (P < 0.05) the Bifidobacterium population. Other bacterial groups enumerated were unaffected with the exception of an increase in the growth of members of the Bacteroides-Prevotella group, Faecalibacterium prausnitzii, and clostridial cluster IX (P < 0.05). Compared to the other substrates, AcGGM promoted butyrogenic fermentation whereas AcAGX was more propiogenic. Although further in vivo confirmation is necessary, these results demonstrate that both AcGGM and AcAGX from lignocellulosic feedstocks can be used to direct the promotion of beneficial bacteria, thus exhibiting a promising prebiotic ability to improve or restore gut health.IMPORTANCE The architecture of the gut bacterial ecosystem has a profound effect on the physiology and well-being of the host. Modulation of the gut microbiota and the intestinal microenvironment via administration of prebiotics represents a valuable strategy to promote host health. This work provides insights into the ability of two novel wood-derived preparations, AcGGM and AcAGX, to influence human gut microbiota composition and activity. These compounds were selectively fermented by commensal bacteria such as Bifidobacterium, Bacteroides-Prevotella, F. prausnitzii, and clostridial cluster IX spp. This promoted the microbial synthesis of acetate, propionate, and butyrate, which are beneficial to the microbial ecosystem and host colonic epithelial cells. Thus, our results demonstrate potential prebiotic properties for both AcGGM and AcAGX from lignocellulosic feedstocks. These findings represent pivotal requirements for rationally designing intervention strategies based on the dietary supplementation of AcGGM and AcAGX to improve or restore gut health.

High Yield of Butanol Production in Repeated Batch Culture Fermentation by Clostridium Acetobutylicum YM1

High Yield of Butanol Production in Repeated Batch Culture Fermentation by Clostridium Acetobutylicum YM1

Modelling the role of microbial p-cresol in colorectal genotoxicity

ABSTRACT Background A greater understanding of mechanisms explaining the interactions between diet and the gut microbiota in colorectal cancer is desirable. Genotoxic microbial metabolites present in the colon may be implicated in carcinogenesis and potentially influenced by diet. Aims We hypothesised that microbial p-cresol is a colonic genotoxin and set out to model potential exposures in the colon and the effects of these exposures on colonic cells. Methods Batch culture fermentations with human faecal inoculate were used to determine the synthesis of p-cresol and other metabolites in response to various substrates. The fermentation supernatants were evaluated for genotoxicity and the independent effects of p-cresol on colonic cells were studied in vitro. Results In batch culture fermentation, supplementary protein increased the synthesis of phenols, indoles and p-cresol, whereas supplementary fructoligosaccharide (FOS) increased the synthesis of short chain fatty acids. The p-cresol was the greatest predictor of genotoxicity against colonocytes in the fermentation supernatants. Spiking fermentation supernatants with exogenous p-cresol further increased DNA damage, and independently p-cresol induced DNA damage in a dose-dependent manner against HT29 and Caco-2 cells and influenced cell cycle kinetics. Conclusions In the colon p-cresol may reach physiologically significant concentrations which contribute to genotoxic exposures in the intestinal lumen, p-cresol production may be attenuated by substrate, and therefore diet, making it a potential modifiable biomarker of genotoxicity in the colon.

Sustainable biosynthesis of curdlan from orange waste by using Alcaligenes faecalis: A systematically modeled approach

This study presents an engineered approach for sustainable biosynthesis of curdlan by Alcaligenes faecalis using orange peels. To confirm the substrate suitability a four step study was organized. Firstly, drying of substrate was carried within temperature range of 60–120 °C, along with the application of moisture diffusion control model. Secondly, fermentation medium was obtained via saccharification and detoxification, releasing highest sugar at 72.34 g/L with phenolics removal of 95–98%. Thirdly, curdlan fermentation was conducted in detoxified orange peel hydrolysate followed by optimization of batch culture fermentation via kinetic modeling using Logistic and Luedeking-Piret equations. In 5 L bioreactor, highest specific growth rate (μm = 0.233/h), highest curdlan production (Pm = 23.24 g/L) and growth associated rate constant (α = 3.403) were achieved. Moreover, the total sugar consumption and conversion rates were 83.27% and 53.20%. Lastly, characterization techniques such as FTIR, NMR, XRD, TGA, HPGPC and EDS were applied to biosynthesized curdlan for qualitative validation.

Human colonic microbiota modulation and branched chain fatty acids production affected by soy protein hydrolysate

SummarySeveral functional properties have been attributed to soy protein hydrolysates (SPHs); however, their gut fermentation needs to be investigated. This study aimed to determine the effect of hypo‐allergic pepsin‐educed SPH on the growth of gut microbiota, SCFA and BCFA production in a pH‐controlled, stirred, batch culture fermentation system compared with commercial SPH and soy protein isolate (SPI). The results showed that all substrates affected gut bacteria. SPH selectively increased the number of lactobacilli (log 9.61 to log 9.84) at 72‐h fermentation, but not for bifidobacteria. This is in accordance with increased BCFAs yield of 15.54 ± 0.00 mm (isobutyrate) and 465.59 ± 1.42 mm (isovalerate). The gut microbial balance index (GMBI) of SPH was 0.23, thus suggesting its gut microbial modulation. Therefore, SPH has potential for modulation or balance of gut microbiota, which may be used as functional ingredient for emerging market of nutraceuticals and nutri‐pharmaceutical industry.

Enhanced rhizosphere competence of Trichoderma viride grown in solid state fermentation on corn cob residue

A process has been disclosed in the present investigation for the production of a formulation of mycopesticide with enhanced shelf life of Trichoderma viride. A comparative evaluation of talc-based and corn cob formulations in the form of seed coat and soil treatment brought to light that the soil treatment significantly enhanced the plant growth performance of pea, green gram and pigeon pea. Among the two formulations, the corn cob formulation proved to be better as the rhizosphere competence of corn cob formulation was superior to that of talc formulation. Our results suggest that corn cob residue is a better substrate owing to high cellulase and spore production. Solid-state fermentation in batch cultures. The biocontrol ability of both formulations was tested against Fusarium in Cajanus species and it was found that the disease incidence was reduced by 86% when the corn cob formulation was given as soil application.

Metabolic retroconversion of trimethylamine N-oxide and the gut microbiota

BackgroundThe dietary methylamines choline, carnitine, and phosphatidylcholine are used by the gut microbiota to produce a range of metabolites, including trimethylamine (TMA). However, little is known about the use of trimethylamine N-oxide (TMAO) by this consortium of microbes.ResultsA feeding study using deuterated TMAO in C57BL6/J mice demonstrated microbial conversion of TMAO to TMA, with uptake of TMA into the bloodstream and its conversion to TMAO. Microbial activity necessary to convert TMAO to TMA was suppressed in antibiotic-treated mice, with deuterated TMAO being taken up directly into the bloodstream. In batch-culture fermentation systems inoculated with human faeces, growth of Enterobacteriaceae was stimulated in the presence of TMAO. Human-derived faecal and caecal bacteria (n = 66 isolates) were screened on solid and liquid media for their ability to use TMAO, with metabolites in spent media analysed by 1H-NMR. As with the in vitro fermentation experiments, TMAO stimulated the growth of Enterobacteriaceae; these bacteria produced most TMA from TMAO. Caecal/small intestinal isolates of Escherichia coli produced more TMA from TMAO than their faecal counterparts. Lactic acid bacteria produced increased amounts of lactate when grown in the presence of TMAO but did not produce large amounts of TMA. Clostridia (sensu stricto), bifidobacteria, and coriobacteria were significantly correlated with TMA production in the mixed fermentation system but did not produce notable quantities of TMA from TMAO in pure culture.ConclusionsReduction of TMAO by the gut microbiota (predominantly Enterobacteriaceae) to TMA followed by host uptake of TMA into the bloodstream from the intestine and its conversion back to TMAO by host hepatic enzymes is an example of metabolic retroconversion. TMAO influences microbial metabolism depending on isolation source and taxon of gut bacterium. Correlation of metabolomic and abundance data from mixed microbiota fermentation systems did not give a true picture of which members of the gut microbiota were responsible for converting TMAO to TMA; only by supplementing the study with pure culture work and additional metabolomics was it possible to increase our understanding of TMAO bioconversions by the human gut microbiota.

In vitro fermentation of copra meal hydrolysate by chicken microbiota.

The aim of this study was to carry out preliminary investigations on the in vitro fermentation selectivity of copra meal hydrolysate (CMH) by chicken gut microbiota. The ileum and cecum contents from three 35-day-old birds were used as inocula. Yeast mannooligosaccharide (yeast-MOS) or α-mannan was selected as a positive control. Batch culture fermentation with fecal bacteria was performed at 42°C for 24h in an anaerobic chamber. Samples were collected at 0, 6, 12, 18 and 24h of fermentation and evaluated using real-time PCR and short-chain fatty acid (SCFA) analysis. Results showed that the medium containing ileum and both CMH and yeast-MOS substrates led to an increase in the growth of the dominant groups as Lactobacillus, Enterobacteriaceae and Enterococcus spp. compared with 0-h fermentation. Campylobacter spp. and Bifidobacterium spp. were not detected in any samples. A significant decrease in Acinetobacter was observed in all substrates tested after 6h of fermentation (P<0.05). Only the sample from CMH fermentation showed a significantly greater reduction in the population of Pseudomonas after 18-h fermentation with ileum content (P<0.05). Propionate was the main fermentation product found in both ileum and cecum fermentation followed by lactate and acetate. CMH can be utilized by ileum and cecum microbial of chickens, and CMH has a generally desirable effect on the microbiota. CMH has the potential for use as a supplementary diet with similar or improved benefits and lower costs compared to commercial prebiotics. Further experiments in animal trials would seem to be justified.

Practical algorithm for stochastic optimal control problem about microbial fermentation in batch culture

How to add glycerol to maximize production of 1,3-propanediol (1,3-PD) is a critical problem in process control of microbial fermentation. Most of the existing works are focusing on modelling this process through deterministic-based differential equations. However, this process is not deterministic, but intrinsically stochastic considering nature of interference. Thus, it is of importance to consider stochastic microorganism. In this paper, we will modelling this process through stochastic differential equations and maximizing production of 1,3-PD is formulated as an optimal control problem subject to continuous state constraints and stochastic disturbances. A modified particle swarm algorithm through integrating the hybrid Monte Carlo sampling and path integral is proposed to solve this problem. The constraint transcription, local smoothing and time-scaling transformation are introduced to handle the continuous state constraints. Numerical results show that, by employing the obtained optimal control governed by stochastic dynamical system, 1,3-PD concentration at the terminal time can be increased compared with the previous results.

In vitro investigation of orange fleshed sweet potato prebiotic potential and its implication on human gut health

Background: Some food ingredients (prebiotics) have been shown to promote a healthy gut by selectively stimulating growth/activity of beneficial gastrointestinal microbes and metabolites such as short chain fatty acids (SCFA) while inhibiting pathogens. Orange fleshed sweet potato (Ipomoea batatas Lam; OFSP) root tuber is a starchy tropical crop and highly nutritious in terms of pro-vitamin A (beta carotene), dietary fibre, and natural sugars, with negligible amount of fats and cholesterol. Purpose of study: The aim of the study was to investigate using simulated human gut system whether OFSP may have prebiotic activity derived from their fibre, resistant starch, and/or the sugars.Methodology: In vitro pH controlled stirred batch culture fermentation system was used to compare the effect on human gut microbiota of four substrates: two varieties of OFSP (SPK 004 and Tainung), FOS and sucrose known for positive prebiotic and non-selective change respectively. The system was inoculated with faecal slurry from six different human healthy donors from different ethical backgrounds, age, and the effectual change recorded over 24 hours by monitoring bacterial counts (total bacteria, Bacteroides and Bifidobacterium) using qPCR molecular technique and SCFA profiles by gas chromatography.Results: The total bacteria count increased by (0.92-1.7 log10) and Bacteroides genus (1.03-1.8 log10) throughout the experimental period but with no significant differences (p&lt;0.05) between the four substrates. However, there were significant differences (p&lt;0.05) in the beneficial Bifidobacterium (1.66-2.66 log10) between the 2 varieties of OFSP and the two controls (FOS and sucrose). The levels of SCFA increased, with acetate as the predominant acid and lactic acid being the least. The OFSP purees elicited high butyric acid levels, which were comparable to those of positive control FOS.Conclusions: The study demonstrated that OFSP purees may have prebiotic potential that can positively modulate gut microbiota by promoting growth of beneficial bacteria, bifidobacterium genus, and stimulating production of SCFA especially butyric acid which is the favourable in human gut health. However, further research using more probiotic and pathogenic microbes in addition to in vivo clinical studies and compositional analysis of OFSP is needed to confirm prebiotic activity. Key words: Orange fleshed sweet potato, prebiotic, human gut microbiota

In vitro fermentability of xylo-oligosaccharide and xylo-polysaccharide fractions with different molecular weights by human faecal bacteria

Xylo-oligosaccharides and xylo-polysaccharides (XOS, XPS) produced by autohydrolysis of the fibre from oil palm empty fruit bunches (OPEFB) were purified using gel filtration chromatography to separate the XOS and XPS from the crude autohydrolysis liquor. Six mixed fractions of refined XOS and XPS with average degree of polymerisation (avDP) of 4–64 were obtained. These were characterised in terms of their composition and size by HPLC, MALDI-ToF-MS (selected fractions) and carbohydrate gel electrophoresis (PACE). They were assessed in batch culture fermentations using faecal inocula to determine their ability to modulate the human faecal microbiota in vitro by measuring the bacterial growth, organic acid production and the XOS assimilation profile. The gut microbiota was able to utilise all the substrates and there was a link between the avDP with the fermentation properties. In general, XOS/XPS preparations of lower avDP promote better Bifidobacterium growth and organic acid production.

553 Effects of pentacyclic triterpenes on in vitro fermentation of bahiagrass hay and a high-grain substrate

Triterpenes are the most representative group of phytochemicals, comprising more than 20,000 recognized molecules. The diversity of triterpenes is highly associated with their broad range of pharmacological effects. Previous studies have demonstrated their anti-microbial potential. Our objective was to determine potential effects of pentacyclic triterpenes (TT) on in vitro batch culture fermentation of bahiagrass hay and a high-grain substrate. A randomized complete block design was used to determine the effects of increasing concentrations of the following TT types: oleanolic acid (OA), ursolic acid (UA) and malsinic acid (MAL) on in vitro fermentation of bahiagrass hay (BGH) or a high-grain substrate (HG) comprised of 43% soybean hulls pellets, 42% corn gluten feed pellets, and 15% peanut hulls (DM basis). Treatments were arranged as a 4 × 3 factorial with 4 doses (0, 4, 25 and 100 mg/L of incubation volume) for each of the 3 TT types. Four ruminally cannulated crossbred steers (362 ± 30 kg BW) were used as ruminal fluid donors. Two steers were fed HG ad libitum, while the other two steers were fed BGH ad libitum for at least 14 d prior to the first incubation day. Each replicate d, 125-mL serum bottles containing 50 mL of a 4:1 buffer:ruminal fluid inoculum and 0.63 g of BGH substrate (DM basis) were incubated for 48 h, and a 2:1 buffer:ruminal fluid inoculum and 0.63 g of HG substrate (DM basis) were incubated for 24 h. Three d (block) of incubation were performed. In vitro OM digestibility (IVOMD) was determined in a separate set of tubes. Data were analyzed using PROC MIXED of SAS with the fixed effect of treatment, and random effect of incubation day. No effect was observed (P>0.05) on total gas production, IVOMD, NH3-N concentration and H2S production. There was no effect (P>0.05) of any TT types or doses on concentrations or molar proportions of VFA. No effect (P>0.05) of TT type or dose were observed on acetate:propionate ratio (A:P). In conclusion, the acid form of the pentacyclic triterpenes did not affect ruminal fermentation of either substrate. Future studies should focus on increasing the solubility of these pentacyclic triterpenes by using a sodium salt.

Impact of pH and butyric acid on butanol production during batch fermentation using a new local isolate of Clostridium acetobutylicum YM1.

The effect of pH and butyric acid supplementation on the production of butanol by a new local isolate of Clostridium acetobutylicum YM1 during batch culture fermentation was investigated. The results showed that pH had a significant effect on bacterial growth and butanol yield and productivity. The optimal initial pH that maximized butanol production was pH 6.0 ± 0.2. Controlled pH was found to be unsuitable for butanol production in strain YM1, while the uncontrolled pH condition with an initial pH of 6.0 ± 0.2 was suitable for bacterial growth, butanol yield and productivity. The maximum butanol concentration of 13.5 ± 1.42 g/L was obtained from cultures grown under the uncontrolled pH condition, resulting in a butanol yield (YP/S) and productivity of 0.27 g/g and 0.188 g/L h, respectively. Supplementation of the pH-controlled cultures with 4.0 g/L butyric acid did not improve butanol production; however, supplementation of the uncontrolled pH cultures resulted in high butanol concentrations, yield and productivity (16.50 ± 0.8 g/L, 0.345 g/g and 0.163 g/L h, respectively). pH influenced the activity of NADH-dependent butanol dehydrogenase, with the highest activity obtained under the uncontrolled pH condition. This study revealed that pH is a very important factor in butanol fermentation by C. acetobutylicum YM1.

In vitro approaches to assess the effects of açai (Euterpe oleracea) digestion on polyphenol availability and the subsequent impact on the faecal microbiota

A considerable proportion of dietary plant-polyphenols reach the colon intact; determining the effects of these compounds on colon-health is of interest. We hypothesise that both fibre and plant polyphenols present in açai (Euterpe oleracea) provide prebiotic and anti-genotoxic benefits in the colon. We investigated this hypothesis using a simulated in vitro gastrointestinal digestion of açai pulp, and a subsequent pH-controlled, anaerobic, batch-culture fermentation model reflective of the distal region of the human large intestine.Following in vitro digestion, 49.8% of the total initial polyphenols were available. In mixed-culture fermentations with faecal inoculate, the digested açai pulp precipitated reductions in the numbers of both the Bacteroides-Prevotella spp. and the Clostridium-histolyticum groups, and increased the short-chain fatty acids produced compared to the negative control. The samples retained significant anti-oxidant and anti-genotoxic potential through digestion and fermentation.Dietary intervention studies are needed to prove that consuming açai is beneficial to gut health.

In vitro ruminal fermentation and methane production of different seaweed species

Seaweeds have potentials as alternative feed for ruminants, but there is a limited knowledge on their nutritive value. Seven seaweed species collected along the coast above the Arctic circle of Norway, both in spring and autumn, were assessed for nutrients and total polyphenols (TEP) content, gas production kinetics and in vitro rumen fermentation in batch cultures of ruminal microorganisms. The seaweeds were three red species (Mastocarpus stellatus, Palmaria palmata and Porphyra sp.), three brown species (Alaria esculenta, Laminaria digitata and Pelvetia canaliculata) and one green species (Acrosiphonia sp.). Additionally, the abundance and diversity of total bacteria, protozoa and archaea in the cultures with the three red seaweeds collected in spring were analyzed by quantitative PCR and PCR-DGGE, respectively. The crude protein (CP) content varied widely. Pelvetia had the greatest (P<0.001) ether extract (EE) content. Non-structural carbohydrates (NSC) content varied from 135 to 541g/kg DM with brown seaweeds having the greatest values. Ash and CP contents were higher in spring than in autumn (P=0.020 and 0.003, respectively), whereas concentrations of EE and NSC were not affected by collecting season (P=0.208–0.341). The TEP values ranged from 1.46 to 50.3mg/g dry matter (DM), and differed (P<0.001) among seaweed species and collecting season, being greater in autumn than in spring. The DM effective degradability (DMED), estimated from gas production parameters for a rumen passage rate of 3.0% per h, ranged from 424 to 652g/kg, the highest values were recorded for Mastocarpus stellatus and Porphyra sp. The lowest DMED values were registered for Pelvetia canaliculata and Acrosiphonia sp. In 24-h incubations (500mg DM), Palmaria palmata had the highest (P<0.05) volatile fatty acids (VFA) and methane production (4.34 and 0.761mmol, respectively) and the lowest (P<0.05) final pH values and acetate to propionate ratios (6.57 and 2.34, respectively). There were no differences (P>0.05) among the other seaweeds in VFA production, but Porphyra sp. had the second highest methane production (P<0.05; 0.491mmol) compared with the other seaweeds (0.361mmol; averaged value). The methane/total VFA ratio was not affected (P>0.05) by either seaweed species or the collection season. Higher final pH (P<0.05) and lower (P<0.05) methane and VFA production, ammonia-N concentrations and DMED values were promoted by the fermentation of seaweed collected in autumn compared with those from spring. Among the red seaweeds, there were no species-specific differences (P>0.05) in the abundance or the diversity of total bacteria, protozoa and archaea. In the PCR-DGGE analysis, samples were separated by the incubation run for all microbial populations analyzed, but not by seaweed species. The results indicate that seaweed species differ markedly in their in vitro rumen degradation, and that samples collected in autumn had lower rumen degradability than those collected in spring.

Production of Acetone, Butanol, and Ethanol (ABE) by Clostridium acetobutylicum YM1 from Pretreated Palm Kernel Cake in Batch Culture Fermentation

The viability of most fermentation processes is very much dependent on the cheap fermentation medium used. Palm kernel cake (PKC) is an abundant biomass generated from the palm oil processing industry that can be used as the carbon source for the growth and production of acetone-butanol-ethanol fermentation (ABE) by Clostridia. In this study, ABE production from the fermentation of PKC using Clostridium acetobutylicum YM1 in a batch culture was conducted. The PKC was subjected to treatment with acids (sulphuric and hydrochloric acids), alkali (sodium hydroxide and alkaline peroxide), enzymatic hydrolysis, and hydrothermal treatment (in autoclave). The sulphuric acid-treated PKC (2% SAPKC) method produced the highest concentration of reducing sugars (30 g/L) compared with the other methods applied. The results showed that increasing the concentration of H2SO4 up to 3% decreased the amounts of generated reducing sugars to 20.4 g/L, which is about 32% less. The fermentation of 1%, 2%, and 3% SAPKC resulted in the production of ABE of 1.07, 5.72, and 3.48 g/L, respectively. This study showed that the pretreatment of PKC improved the content of fermentable sugars and subsequently enhanced the production of ABE by C. acetobutylicum YM1. This study also revealed that PKC can be regarded as a potentially low cost substrate for ABE fermentation.

Removal of Ochratoxin A in Saccharomyces cerevisiae Liquid Cultures

The capacity for removal of ochratoxin A (OTA) during alcoholic fermentation was evaluated in batch systems with one commercial strain and one wild strain of Saccharomyces cerevisiae. Batch alcoholic fermentations were carried out in yeast extract-malt extract broth (YM) medium, with 18.0% glucose and OTA added to final concentrations of 3.48 and 4.95 ng/mL respectively. The removal capacity of each yeast strain was examined after completion of fermentation in batch culture and after extended contact with yeast biomass. The removal capacity of the yeast strains was also examined in stationary phase cultures. Stationary phase yeasts were studied with biomass harvested from the stationary phase of anaerobic fermentation, by incubation in phosphate buffer, with the addition of 5.00 ng/ mL of OTA. Removal studies with stationary phase cells were performed with viable and non-viable cells inactivated with Na-azide. The study showed that in growing phase cultures, OTA removal was significant only after extended contact with yeast biomass; up to 29.7% and 25.4% for wild yeast ZIM 1927 and commercial yeast Lalvin EC-1118 respectively, but not during alcoholic fermentation. In stationary phase cultures, viable and non-viable cells were not significantly different in OTA removal from the medium. This demonstrated that OTA was not metabolised, but possibly adsorbed by the yeast cells. The presence of OTA in synthetic media influenced yeast metabolism, causing the production of higher volatile acidity by 0.08 and 0.13 g/L for Lalvin EC-1118 and ZIM 1927 respectively, and lower concentrations of reducing sugar, by 0.32 g/L, but only for ZIM 1927.

In vitro fermentation of different fructo-oligosaccharides by Bifidobacterium strains for the selection of synbiotic combinations

The use of selected probiotics, prebiotics and/or synbiotics, constitute an interesting dietary strategy for intestinal microbiota modulation in case of dysbiosis. Species of the genus Bifidobacterium are among the most currently used probiotics for human consumption since they have shown beneficial effects in the prevention and treatment of some disorders. Bifidobacteria are saccharolytic microorganisms, but their ability to use different carbohydrates varies among strains. In this study, we investigate the utilization of three prebiotic substrates (two different short-chain fructo-oligosaccharides [scFOS] and inulin) by strains of Bifidobacterium, in order to determine the synbiotic potential of the different probiotic/prebiotic combinations. Batch culture fermentations from six Bifidobacterium strains (Bifidobacterium longum IPLA20021, B. longum IPLA20022, Bifidobacterium animalis IPLA20031, B. animalis IPLA20032, B. animalis IPLA20020 and B. animalis Bb12) were carried out in the presence of inulin or scFOS (Synergy or Actilight), or glucose, as carbon source. Bifidobacteria levels were quantified by plate counting. The pH and production of organic acids in the different batch-culture fermentations were also determined. Our results showed that all the studied strains of B. animalis and B. longum were able to utilize scFOS but not inulin. The use of scFOS as carbon source affected the pattern of metabolite's production, when compared with cultures carried out in glucose, particularly in the case of B. longum. The results indicated that the scFOS are well suited to be used in combination with B. animalis or B. longum strains for the development of synbiotic foods or food supplements.