Evaluation of the yield,molar mass of exopolysaccharides,and rheological properties of gels formed during fermentation of milk by Streptococcus thermophilus strains St-143 and ST-10255y

The yield and chemical structures of exopolysaccharides (EPS) produced by many strains of Streptococcus thermophilus have been characterized. However, the kinetics (or production profile) for EPS during milk fermentation is not clear. In this study, we investigated whether any differences existed in the yield and molar mass of EPS when milk was fermented at the same acidification rate by 2 strains of S. thermophilus (St-143 and ST-10255y). The type of EPS produced by these 2 strains is different. Milk samples were analyzed for EPS concentration every 30 min during a fermentation period of 270 min (final pH 4.5) by using a modified quantification method, which was faster and validated for its recovery of added EPS. Rheological properties of milks during fermentation were also analyzed using small-strain dynamic oscillatory rheology. For the determination of molar mass, EPS extracts were isolated by ultrafiltration of whey obtained during fermentation of milk to pH values 5.2, 4.9, 4.7, and 4.5, and molar mass was analyzed using size-exclusion chromatography–multi-angle laser light scattering. During fermentation, both strains appeared to start producing significant amounts of EPS after about ～150 min, which corresponded to pH ～5.3, which was close to the point of gelation. During the remainder of the fermentation process (150–270 min), the EPS concentration from strains St-143 and ST-10255y significantly increased from 30 to 72 mg/L and from 26 to 56 mg/L, respectively. The quantity of EPS recovered by our modified method was estimated to represent ～60% of the total EPS added to milk. The molar mass of EPS produced by both strains appeared to slightly decrease during fermentation. At pH 5.2, EPS from St-143 and ST-10255y had molar masses of 2.9 × 10⁶ and 1.4 × 10⁶ g/mol, respectively, which decreased to 1.6 × 10⁶ and 0.8 × 10⁶ g/mol, respectively, when the pH of milk was 4.5. Distinct differences were apparent in the rheological properties of gels fermented by the 2 strains. At the end of fermentation, St-143 fermented milk had weaker gels with storage modulus (G′) value at pH 4.6 of 26 Pa, whereas gels made with ST-10255y were stiffer with a G′ value at pH 4.6 of 82 Pa. For St-143 gels, maximum loss tangent (LT_max) values were higher (0.50) and occurred earlier (at a higher pH value) than the LT_max values (0.46) for gels from ST-10255y strain. Because the fermentation conditions were identical for both strains, the observed changes in rheological properties could be due to the differences in chemical structures and molar mass of the EPS produced by these 2 S. thermophilus strains.     

Pilot-scale production of exopolysaccharide from Leuconostoc pseudomesenteroides XG5 and its application in set yogurt

Exopolysaccharide from Leuconostoc pseudomesenteroides XG5 (XG5 EPS) is a linear dextran that is built by glucose units via α-1,6 glycosidic bond. The primary objective of this study was to investigate the yield of XG5 EPS and its application in set yogurt. In laboratory scale, the culture conditions of XG5 EPS production were optimized using the L9 (3³) orthogonal test. Here, the optimized yield of XG5 EPS was 26.02 g/L under the conditions of 100 g/L sucrose, initial pH 7.0, 25°C incubation, and 100 rpm for 36 h in a shaking flask. Based on the optimized parameters of laboratory scale, a pilot fed-batch fermentation was performed in a 50-L bioreactor with an adjusted agitation speed of 20 rpm. The XG5 EPS yield reached 40.07 g/L in fed-batch fermentation, which was 54% higher than that achieved in laboratory scale. In addition, XG5 EPS was added into set yogurt to investigate its effect on the stability of set yogurt. Our data demonstrated that the XG5 EPS improved the water-holding capacity, texture profile, and viscosity of set yogurt during cold storage compared with the controls. In particular, addition of 0.5% XG5 EPS increased the structure of 3-dimensional network of set yogurt, which eventually improved the physical stability of the set yogurt. Overall, this study provided new insights for exploring the pilot scale production and application of dextran.     

Optimization for the maximum bacteriocin production of Lactobacillus brevis DF01 using response surface methodology

To enhance the production of bacteriocin DF01, produced by Lactobacillus brevis DF01, cultivation conditions and medium composition were optimized by using response surface methodology (RSM). The selected 5 factors based on MRS medium were glucose, yeast extract, MgSO₄, temperature, and initial pH. Fractional factorial design (FFD) was effective in searching for the main factors. By a 2^5?1 FFD, glucose, yeast extract, and initial pH were found to be significant factors and had positive effects on bacteriocin production. The effects of the 3 main factors on bacteriocin production were further investigated by a central composite design (CCD). RSM revealed that the maximum bacteriocin production was achieved at yeast extract concentration of 14.56 g/L, glucose concentration of 28.95 g/L, and initial pH of 6.8. After RSM, the titer of bacteriocin was increased by 4-fold.     

产胞外多糖泰山羊肚菌液体培养条件的优化

对泰山羊肚菌产胞外多糖 (exopolysaccharides,EPS)的液体培养基组成和发酵条件进行了优化 ,最适碳源是葡萄糖 ,最适氮源是NH4NO3 。正交试验确定最佳培养基组成为麸皮 2 0 0 g/L ,葡萄糖 30 g/L ,NH4NO3 1g/L ,KH2 PO42 g/L ,MgSO4·7H2 O 1 5 g/L。最佳发酵条件为 2 5℃ ,起始 pH值 6 5 ,装液量 10 0mL/瓶 ,接种量10 % ,摇床转速 2 0 0r/min ,发酵时间 4d。在此条件下 ,其胞外多糖含量 (2 135 4 4 1mg/L)比对照(14 5 4 6 39mg/L)提高了 4 6 8%。     

Submerged culture conditions for mycelial yield and polysaccharides production by Lyophyllum decastes

The effects of various carbon and nitrogen sources, their concentrations, initial pH and fermentation duration on the production of mycelia in terms of dry weight, exo-polysaccharide (EPS) and inner polysaccharide (IPS) by Lyophyllum decastes, a culinary-medicinal mushroom, were investigated in shake-flask cultures. Lactose, glucose and fructose were the top three best carbon sources for mycelial growth with corresponding yields of 6.73 g/l, 6.36 g/l and 6.10 g/l, respectively. Glucose was the best for production of EPS and IPS with 1.65 g/l and 317 mg/g dry mycelia, respectively. Maltose also performed well for EPS production. Yeast extract was the best nitrogen source for the production of mycelia (7.03 g/l) and IPS (325 mg/g dry mycelia), whereas EPS was improved further by increasing the yeast extract concentration (2.46 g/l at 2%). Similarly, initial pH 7 and 8 were best for polysaccharides production (EPS 1.73 g/l and IPS 320 mg/g) and mycelial growth (7.10 g/l), respectively. Maximum mycelial growth peaked at 15 days of cultivation whereas polysaccharides peaked at 10 days, and then tapered off. A concentration of glucose 3% and yeast extract 1% (mycelial yield and IPS) were found to be a suitable condition for submerged culture.     

Statistical modeling and optimization for exopolysaccharide production by Lactobacillus confusus in submerged fermentation under high salinity stress

In the last two decades, many studies have been reported that a high concentration of NaCl suppresses exopolysaccharide (EPS) production in lactic acid bacteria. In the present study, however, the enhancement of EPS production by Lactobacillus confusus under high salinity stress in submerged fermentation was demonstrated using response surface methodology via a full factorial design. Under the optimized conditions of 3.33% NaCl, 20 g/L sucrose, and 35 h of incubation, the EPS yield was 10.87 g/ L with 178% higher than the maximum yield (6.12 g/L of EPS) produced from the modified MRS medium without NaCl. Biomass production was independent of EPS production. A high yield of biomass was obtained in the culture with 0.23% NaCl. This results indicate that high salinity stress by NaCl can enhance EPS production in submerged fermentation in uncontrolled pH cultivations by inducing the production of cell-associated dextransucrase.     

KINETICS OF BIOETHANOL PRODUCTION FROM WHEAT MILLING BY-PRODUCTS

An overview of the potential application of wheat milling by‐products as substrate for bioethanol production is presented. In order to select a suitable microorganism, model fermentations were conducted using glucose and dry baker's yeast. The overall ethanol yield was nearly stable (ca. 0.35 g/g), independent of mash glucose concentration; mashes with 100 g glucose/L resulted in an overall ethanol productivity of 3.48 g/L·h. Slurries containing low‐grade wheat flour (LG) (100, 200 or 300 g/L) were used for simultaneous saccharification and fermentation (SSF) with Zymomonas mobilis. Fermentation performance was evaluated based on ethanol concentration (P), productivity (Q_v), yield (Y_P/S), production rate (Q_p) and glucose consumption rate (Q_s). Mashes containing 200 g LG/L produced about 52 g ethanol/L, with Q_vof 2.17 g/L·h. Based on the relatively high fermentation rate of LG, reaching peak ethanol productivity within ca. 9 h of SSF, considerable savings on fermentation time was achieved. Using Z. mobilis for LG fermentation, P was about 30% higher than that obtained with Saccharomyces cerevisiae.     

Optimal production of exopolysaccharide by Bacillus licheniformis KS-17 isolated from kimchi

A strain producing exopolysaccharide (EPS) with strong hydroxyradical scavenging activity and antityrosinase activity has been isolated previously. However, the EPS production rate of the strain (7.3 g/L) was relatively low to utilize it at the industrial level. Therefore, in this study, optimal medium and fermentation conditions were examined to enhance EPS production by Bacillus licheniformis KS-17. Maximum EPS production was obtained in medium with 125 g/L sucrose, 30 g/L ammonium sulfate, and 10 mM calcium chloride without a phosphate source at 37°C for 5 days with the initial pH adjusted to 8.5. Under optimal culture conditions, EPS was produced at up to 27.2 (at 5 days) vs. 12.0 g/L (at 7 days under basal conditions), which was 2.3 times greater and in a shorter time than the production yield possible without optimizing conditions.     

Evaluation of the conditions of carotenoids production in a synthetic medium by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor

This work studied the cultivation conditions for the production of carotenoids by Sporidiobolus salmonicolor (CBS 2636) in a bioreactor. A Plackett–Burman design was used for the screening of the most important factors, followed by a complete second order design, to maximise the concentration of total carotenoids. The maximum concentration of 3425.9 μg L^?1 of total carotenoids was obtained in a medium containing 80 g L^?1 glucose, 15 g L^?1 peptone and 5 g L^?1 malt extract, with an aeration rate 1.5 vvm, 180 r.p.m., 25 °C and an initial pH of 4.0. Fermentation kinetics showed that the maximum concentration of total carotenoids was reached after 90 h of fermentation. Carotenoid bio‐production was partially associated with cell growth. The specific carotenoid production (Y_P/X) was 238 μg carotenoids/g cells, whereas Y_P/S (substrate to product yield) was 41.3 μg g^?1. The specific growth rate (μ_x) was 0.045 h^?1. The highest cell and total carotenoid productivity were 0.19 g L^?1 h^?1 and 56.9 μg L^?1 h^?1, respectively.     

The effect of optimized carbon source on the synthesis and composition of exopolysaccharides produced by Lactobacillus paracasei

This study aimed to predict the optimal carbon source for higher production of exopolysaccharides (EPS) by Lactobacillus paracasei TD 062, and to evaluate the effect of this carbon source on the production and monosaccharide composition of EPS. We evaluated the EPS production capacity of 20 strains of L. paracasei under the same conditions. We further investigated L. paracasei TD 062, which showed the highest EPS-producing activity (0.609 g/L), by examining the associated biosynthesis pathways for EPS. Genomics revealed that fructose, mannose, trehalose, glucose, galactose, and lactose were carbon sources that L. paracasei TD 062 could use to produce EPS. We identified an EPS synthesis gene cluster that could participate in transport, export, and sugar chain synthesis, and generate 6 sugar nucleotides. Experimental results showed that the sugar content of the EPS produced using fermentation with the optimized carbon source (fructose, mannose, trehalose, glucose, galactose, and lactose) increased by 115%. Furthermore, use of the optimized carbon source changed the monosaccharide content of the associated EPS. The results of enzyme activity measurements showed significant increases in the activity of 2 key enzymes involved in the glycoside synthesis pathway. Our study revealed that optimizing the carbon source provided for fermentation not only increased the production of EPS, but also affected the composition of the monosaccharides by increasing enzyme activity in the underlying synthesis pathways, suggesting an important role for carbon source in the production of EPS by L. paracasei TD 062.     

Effect of nutritional and environmental conditions on the production of exo-polysaccharide of Agaricus brasiliensis by submerged fermentation and its antitumor activity

Mushrooms have become attractive as a functional food and as a source for the development of drugs and nutraceuticals. A. brasiliensis is considered as the best among them. There are several published works on the fruiting bodies of this mushroom, showing high antitumor activity. The mycelium polysaccharide and exo-polysaccharide (EPS) of this mushroom also demonstrated a strong antitumor action. However, there is little information available in the literature about the optimization of physical and chemical conditions for production of EPS by submerged fermentation (SmF). After initial screening from the five strains available in the LPB Culture Collection, A. brasiliensis LPB 03 was selected for EPS production in SmF. Studies on supplementation of a carbon source to the medium revealed that sucrose was most effective for EPS production. Yeast extract was the best for EPS among the inorganic and organic nitrogen sources tested. The factorial experiment demonstrated that a temperature of 30 °C and a pH of 6.1 were the best for the EPS production. Glucose 10 g/l, yeast extract 3 g/l, K₂HPO₄ 0.6 g/l and MgSO₄ 0.3 g/l were most suitable for the EPS production. Maximum EPS production was obtained in the static condition with manual agitation two times per day after a 1-week culture (382 mg/l), increasing by 78.62% more than that before optimization. In the bioreactor with aeration and agitation, the maximum EPS was produced at 321.2 mg/l. The EPS of A. brasiliensis produced in SmF showed strong inhibition against Sarcoma 180 in mice, reaching 72.19% inhibition compared with the control group. Furthermore, 50% of mice in the test group demonstrated total tumor regression.     

Physicochemical characterisation of the exopolysaccharides of Streptococcus thermophilus ST‐143

Exopolysaccharides (EPS) excreted by lactic acid bacteria used in the dairy environment are either liberated into the medium (free EPS) or remain attached to the cells (capsular EPS). After batch fermentation of Streptococcus thermophilus ST‐143 at pH 6.0, three EPS fractions were isolated and purified: free EPS (EPS_f), capsular‐derived EPS (EPS_cd) and the entire total EPS (EPS_total). EPS_f and EPS_cd were uncharged, consisted of rhamnose, galactose, glucose and N‐glucosamine in different amounts and showed a molecular mass of 2.6 × 10⁶ Da (EPS_f) and, in case of EPS_cd, 7.4 × 10³ Da and 1.4 × 10⁵ Da in a ratio of 9: 1. Intrinsic viscosity of the polymers was 1.14 mL mg^?1 (EPS_f), 0.06 mL mg^?1 (EPS_cd) but 1.23 mL mg^?1 for EPS_total. The higher functionality of the entire EPS that was also observed in shear rheology measurements indicates that the presence of small capsular‐derived EPS facilitates the entanglement of the larger polymeric carbohydrates of the EPS_f fraction.     

Influences of pH and electrolyte on the rheological properties of aqueous solution of exopolysaccharide secreted by a deep-sea mesophilic bacterium

The effects of pH and electrolytes (NaCl and CaCl₂) on the viscosity, dynamic modulus (storage modulus G′ and loss modulus G″) and thixotropy of aqueous solution of new exopolysaccharide (SM-A87 EPS) secreted by a deep-sea mesophilic bacterium Wangia profunda SM-A87 were investigated. It was found that at pH 3–12 and the electrolyte concentration of 0.1–80 g/L, the SM-A87 EPS solution showed very weakly pH-dependent and salt-dependent rheological properties and dominant elastic behavior. These properties were considered to arise from the hyper-branched structure of the SM-A87 EPS molecules through the comparison of rheological properties of SM-A87 EPS solutions with those of partially hydrolyzed polyacrylamide ones at different electrolyte concentrations. Such good pH stability and salt resistance of the SM-A87 EPS solution indicate that SM-A87 EPS has great potential for application as food additive, thickener, stabilizer and enhanced oil recovery system. In the studied conditions, 2 g/L SM-A87 EPS solution showed positive thixotropy. The relative thixotropic recovery decreased with pH increasing and the electrolyte concentration decreasing.     

Factors influencing ethanol production rates at high-gravity brewing

The aim of this work was to investigate the influences of different fermentation factors on ethanol production rates by Saccharomyces cerevisiae (lager strain), at high-gravity brewing using response surface methodology. An empirical linear polynomial model was developed to describe the behaviour of the dependent variable as a function of significant factors. The resultant functional relationship in terms of coded values for predicting ethanol production rates was: Y=0.421+0.155X₂+0.0575X₂X_3, where Y represents the ethanol production rate (g/lh), and X₂ and X₃ are coded levels for fermentation temperature and nutrient supplementation, respectively. Patterns of yeast growth, decrease in wort gravity and ethanol production were also evaluated at the maximum ethanol production rate (0.694 g/lh). It was concluded that higher ethanol production rates could be achieved by increasing fermentation temperature and supplementing high-gravity worts with yeast extract, ergosterol and Tween 80.     

Optimization of medium on exopolysaccharides production in submerged culture of Cordyceps militaris

Statistical experimental design strategy (SES) was applied to optimize the medium for the exopolysaccharides (EPS) production of Cordyceps militaris by submerged culture in shaker flask. A significant influence of the glucose and peptone on the EPS production was first evaluated by using a Plackett-Buman design. Then, steepest ascent method was employed to approach the experimental design space. Last, these factors were further optimized using central composite design (CCD) and response surface methodology (RSM). A quadratic model was found to fit the EPS production. The optimum values of the tested variables for the production of EPS were 48.67 g/L glucose, 12.56 g/L peptone, 1 g/L KH₂PO₄, 10 g/L yeast extract, and 0.5 g/L MgSO₄·7H₂O. Under optimization of culture conditions, the EPS production was enhanced from 0.78 to 1.96 g/L. In comparison with that of original culture conditions, 2.5 fold increase was obtained.     

丙酮丁醇梭菌发酵条件优化研究

以P2培养基为基础组分,分别通过改变初始葡萄糖浓度、初始酵母膏浓度以及初始pH值,研究这3个单因素对丁醇发酵的影响,确定了培养基的较佳条件:初始葡萄糖浓度60g/L、初始酵母膏浓度3g/L、初始pH值6.8.此外,采取接种量5％、发酵温度37℃、发酵时间72h,可使总溶剂浓度(丁醇、丙酮、乙醇)达到13.52g/L,其中丁醇、丙酮、乙醇浓度分别为8.83g/L、3.90g/L和0.79g/L,丁醇比例为65.31％.糖丁醇转化率为21.1％(平均值),糖总溶剂转化率为31.3％(平均值).     

Growth and exopolysaccharide yield of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081 in batch and continuous bioreactor experiments at constant pH

Some Lactobacillus delbrueckii ssp. bulgaricus strains are able to synthesize exopolysaccharides (EPS) and are therefore highly important for the dairy industry as starter cultures. The aim of this study was to investigate the nutritional requirements for growth and EPS production of Lactobacillus delbrueckii ssp. bulgaricus DSM 20081. A medium was developed from a semi-defined medium (SDM) in which glucose was replaced by lactose and different combinations of supplements (nucleobases, vitamins, salts, sodium formate and orotic acid) were added. Constant pH batch fermentation with the modified medium resulted in an EPS yield of approximately 210 mg glucose equivalents per liter medium. This was a 10-fold increase over flask cultivation of this strain in SDM. Although not affecting cell growth, the mixture of salts enhanced the EPS synthesis. Whereas EPS production was approximately 12 mg/g dry biomass without salt supplementation, a significantly higher yield (approximately 20 mg/g dry biomass) was observed after adding the salt mixture. In continuous fermentation, a maximal EPS concentration was obtained at a dilution rate of 0.31/h (80 mg EPS/L), which corresponded to a specific EPS production of 49 mg/g dry biomass.     

蚕蛹粉对红酵母发酵生产类胡萝卜素的影响

研究蚕蛹粉浓度、葡萄糖浓度、培养时间以及发酵液初始pH对一株红酵母发酵生产类胡萝卜素的影响.单因素实验结果表明:添加7.5 g/L的蚕蛹粉、葡萄糖浓度为30 g/L,发酵时间4 d,发酵液初始pH 7.0时,类胡萝卜素产量达到最高值14.51 mg/L,较对照组提高46.9％;通过正交实验进一步优化培养条件,结果表明:...     

The effects of particle size on the physicochemical properties of optimized astaxanthin-rich Xanthophyllomyces dendrorhous-loaded microparticles

To improve the entrapment efficiency (EE) of astaxanthin-rich Xanthophyllomyces dendrorhous (ASX)-loaded calcium alginate gel (ASX-CAG) microparticles, we used a response surface methodology to optimize preparation conditions including the ratio of ASX to total material (X₁), alginate concentration (X₂), and CaCl₂ concentration (X₃). The EE and the mean size of the ASX-CAG microparticles were 76.7 g/100 g and 210.26 μm, respectively, after preparation under optimal conditions: 24 g ASX/100 g total material, 1.0 g/100 g alginate, and 200 mmol/L CaCl₂. The effects of particle size on different characteristics were evaluated with increasing microparticle size; an increase in microparticle size significantly increased EE and the antioxidant activity of ASX, but resulted in a decrease in the release of entrapped ASX. Most importantly, the lipid peroxidation inhibitory activity of encapsulated ASX (55.1%) was significantly higher and longer-lasting than that of non-encapsulated ASX (40.5%) after 36 h of storage as determined using the thiobarbituric acid method.     

Exopolysaccharide production by Pediococcus damnosus 2.6 in a semidefined medium under different growth conditions

Ropy Pediococcus damnosus (strain 2.6) was used for production of exopolysaccharide (EPS) in a semidefined medium. From a kinetic point of view, an experiment conducted in SMD medium containing 30 g l(-1) glucose and 5 g l(-1) Bacto casamino acids (Difco Laboratories, Detroit, MI), without pH control, showed that EPS production took place mainly during the growth phase. The viscosity of the cultures developed in parallel to the EPS synthesis until 94 h of incubation; after 200 h of fermentation, viscosity decreased. The effect of glucose, Bacto casamino acid concentrations and temperature on growth and EPS production was determined by using a full factorial design. Within the domain of experimental conditions considered, the concentration of glucose and Bacto casamino acids has a significant effect on the production of exopolysaccharide. The incubation at 12 degrees C produced a prolonged lag phase and due to the lower growth yield, higher specific EPS production was found at this temperature. At 25 degrees C the EPS production was mainly enhanced by the increase in glucose concentration. The increase in nitrogen concentration from 5 to 15 g l(-1) did not yield greater EPS production. However, at 12 degrees C optimal EPS production was obtained when both higher glucose and nitrogen concentrations were used.