Downstream processing of pullulan from fermentation broth

pullulan, a water soluble extracellular polysaccharide, was produced by downstream fermentation employing the strain Aureobasidium pullulans. To obtain pure biopolymer from the fermentation broth, it is necessary to harvest cells, heat the broth, remove the melanin pigments co-produced during fermentation, concentration, precipitate and dry. Centrifugation of the fermentation broth at 10,000 rpm for 15 min gave cell pellets that were discarded and a green–black supernatant containing melanin pigment was subjected to the heat treatment at 80 °C for 20 min in order to remove the protein in the fermentation broth. The supernatant was demelanized by oxidation with hydrogen peroxide, concentrated under vacuum, precipitated with ethanol and dried at 60 °C for 30 min. This procedure produced high purity pullulan that was comparable in color and texture to the commercial samples.     

Influence of pH on the production of enterocin 1146 during batch fermentation

The production of enterocin 1146, a bacteriocin from Enterococcus faecium DPC1146, was studied during batch fermentation at pH 5, 5.5, 6 and 6.5. The bacteriocin was produced throughout the growth of the micro-organism, showing primary metabolite kinetics. Bacteriocin production stopped at the end of growth and was followed by a decrease in activity due primarily to adsorption on the cells of the producer. The optimal pH for enterocin 1146 production was 5.5, because of higher bacteriocin yield per unit of biomass and slower adsorption/degradation, while optimal pH for growth was between 6.0 and 6.5.     

二价阳离子对短梗霉多糖发酵的影响

就二价阳离子对短梗霉多糖和黑色素的影响进行了分析和研究。结果表明 ,二价阳离子对短梗霉多糖的合成和黑色素的形成均有较大的影响。通过对培养基中二价阳离子含量和种类的控制不仅可以抑制细胞黑色素的形成 ,而且还保持了很高的多糖发酵水平 ,在 30L生物反应器中短梗霉多糖的产量和转化率分别达到了 59 8g/L和 61 5%。     

3-羟基丙酸分批发酵过程中的pH控制策略研究

本文利用重组大肠杆菌以甘油为底物发酵合成3．羟基丙酸,考察了不同pH对3．羟基丙酸产量及菌体生长的影响,发现在pH6．5条件下,细胞比生长速率达到最大值,延迟期也相对较短;而pH7．0有利于3-羟基丙酸的合成,控制pH7．0可以使3-羟基丙酸产量达到7．39g／L。基于不同pH条件下对细胞比生长速率和3-羟基丙酸比生成速率的分析,提出3．羟基丙酸分批发酵过程中的pH控制策略,即在发酵过程前5h将pH控制在6．5,5h～15h控制pH为7．0,此时有利于细胞生长;而后在15h-25h控制pH为7．5,25h后控制pH为7．0,从而使细胞具有较高的3．羟基丙酸比合成速率。在此控制策略下经过34h发酵3-羟基丙酸的终产量达到8．76g／L,比pH7．0条件下的3-羟基丙酸产量提高了18．54％。     

Product patterns of non-axenic sucrose fermentation as a function of pH

Summary The product patterns were analyzed for sucrose, fermented by non-axenic continuous cultures, at different pH-values. At pH 4.0–4.5, ethanol was the dominant metabolite. Butyric acid dominated between pH 4.5 and 5.0 while a mixed volatile acid pattern occured at higher pH-values. Hydrogen gas production appeared to be associated with butyric acid and never represented more then 6.5 % of the reducing equivalents of the feed.     

Influence of impeller speed upon the pullulan fermentation

Summary The effect of impeller speed on pullulan production and the morphology ofAureobasidium pullulans in batch culture was studied. Pullulan production and the percentage of yeast cells in the culture rose with impeller speed, as did molecular weight of the polysaccharide.     

Effect of nitrogen source on pullulan production by Aureobasidium pullulans grown in a batch bioreactor

Pullulan production by Aureobasidium pullulans ATCC 201253 using selected nitrogen sources was studied in a medium using corn syrup as a carbon source. Independent of the corn syrup concentration present, the use of corn steep liquor or hydrolysed soy protein as a nitrogen source instead of ammonium sulphate did not elevate polysaccharide production by ATCC 201253 cells grown in an aerated, batch bioreactor containing 4 litres of medium. Pullulan production on corn steep liquor or hydrolysed soy protein as a nitrogen source became more comparable as the concentration of corn syrup was increased. Cell weights after 7 days of growth on any of the nitrogen sources were similar. The viscosity of the polysaccharide on day 7 was highest for cells grown on ammonium sulphate and 12.5% corn syrup. The pullulan content of the polysaccharide elaborated by ammonium sulphate-grown cells on day 7 decreased as the corn syrup level rose in the medium while the pullulan content of polysaccharide produced by cells grown on corn steep liquor or soytone generally increased.     

Production of pullulan by a fed-batch fermentation

Summary In pullulan production from sucrose byAureobasidium pullulans, a sugar concentration higher than 5% (w/v) inhibited cell growth and the production of exopolysaccharide. By a fed-batch fermentation, the inhibitory effects of the high sugar concentration were overcome and 58.0 g/1 of exopolysaccharide were obtained from 10% sucrose.Abbreviations m, n relationship parameters for the growth and non-growth associated product formation - X, Xmax biomass and maximum biomass concentration (g cell/1) - P product concentration (g exopolysaccharide/1) - specific growth rate of cell (hr^–1)     

Effect of the pH of the medium on DNA conformation

The hydrodynamical and optical properties of DNA were investigated in the wide-range of pH by the methods of streaming birefringence, viscometry and spectrophotometry for the different ionic strengths of environment. The measurements of the intrinsic viscosity as a function of pH allow us to determine the compactization of protonated DNA without the destruction of double-helical conformation. This transition is accompanied by a decrease in the optical anisotropy of DNA and the coefficient of molar extinction E260 (P). The increase of volume and persistence length of DNA was observed in the alkaline range of pH. Analyses of experimental data lead to an assumption that the predominant cause of these effects is the change of flexibility of DNA as a result of ionization of its bases. The data obtained were compared with those for polycationic molecules.     

Effect of glucose flow on the acetone butanol fermentation in fed batch culture

Summary Clostridium acetobutylicum was grown in fed-batch cultures at different feeding rates of glucose. The sugar converted to butanol and acetone increased with increasing the glucose flow, on the contrary the conversion to butyric acid was highest at slow glucose feeding rate. The acetic acid concentration was constant at the different flows of glucose. The solventogenesis was not inhibited at high flow of sugar.     

Effect of environmental pH on the fermentation balance of Lactobacillus reuteri

The influence of environmental pH on the regulation of glucose catabolism by Lactobacillus reuteri was examined in anaerobic batch cultures. Under acidic conditions both glucose consumption and end-products formation were low. Maximum biomass was reached at pH 5·0, with a specific growth rate of µ= 0·78 h^-1. The shift in pH values from 4.3 to 6.5 reflected an increase in glucose uptake as well as in the yield ( Y _p/x) of acetate, lactate and ethanol after 12 h of incubation. Ethanol was the major metabolite produced at all pH values assayed.     

Effect of pH on growth of mixed cultures in batch reactor

This work has studied the effect of pH on specific growth rate mu, yield factor Y, and specific substrate consumption rate U for two mixed microbial populations (sludge A and B) in a batch reactor with a limiting substrate (phenol). The mathematical analysis of mu and U as a function of pH has been applied not only to the results of the present work but also to other published data. On the basis of the results obtained, the following remarks can be made: (a) The effect of pH on bacterial activity differed for the two sludges; (b) variations in pH of one unit more or less than optimum pH can give rise to appreciable variations in mu and U; and (c) with regard to the mixed populations used in this study, the mu or U and pH data for the range investigated can be suitably described by a parabolic relation.     

Optimization of pH for high molecular weight pullulan

Summary Pullulan is a polysaccharide produced by Aureobasidium pullulans. In this study, the effect of pH on the molecular weight of pullulan was investigated. High concentration of pullulan was obtained when initial pH was 6. Pullulan having molecular weight of 500,000–600,000 was produced at initial pH of 3.0, while pullulan with molecular weight of 200,000–300,000 was produced at pH above 4.5. To obtain high molecular weight pullulan with high concentration, pH was initially controlled at pH 6, followed by pH shift from pH 6 to pH 3. Transition of pH at 2 days of fermentation was observed to be optimum. Higher molecular weight pullulan was also obtained when sucrose concentration was 50 g/l compared to the result obtained at initial sucrose concentration of 20 g/l. Sucrose concentration and pH of the fermentation broth seem to be important parameters in obtaining high molecular weight of pullulan.     

调控pH提高分批发酵赤霉素GA3产量

为高效率发酵生产GA₃,对藤仓赤霉菌发酵过程pH进行优化调控研究。采用5L全自动发酵罐,在pH 3.0-5.0条件下,对藤仓赤霉菌菌丝生长及GA₃产量的影响进行了考察,实验数据表明：在pH 4.0条件下,菌比生长速率可获最大值,为0.395/h;而pH 3.0条件下,GA₃比生成速率最大,达到4.43mg/(g?h)。基于不同pH条件下,对菌比生长速率、得率、GA₃比生成速率的影响,提出GA₃分批发酵过程中的pH调控策略,即：0-20h,pH自然;20-50h,pH 4.0;50-80h,pH 3.0-3.5;80h后控制pH为3.5-4.0。在此控制模式下,经过196h发酵GA₃的终产量达到2 224mg/L,GA₃产率44.5mg/g,GA₃生产强度0.242mg/(L?h),分别比不控制pH条件下发酵的数值增长了7.75%、7.74%、8.04%,表明该pH控制策略能增进GA₃发酵生产效率。     

Calculation of fermentation parameters from the results of a batch test taking account of the volume of biomass in the fermenting medium

The values of fermentation parameters calculated from the measured concentrations of substrates and/or products may be significantly affected by the volume of biomass in the fermenting medium. Corrections proposed in this paper should be evaluated and, depending on their magnitude, considered in order to obtain more representative results.     

Calculation of fermentation parameters from the results of a batch test taking account of the volume of biomass in the fermenting medium

The values of fermentation parameters calculated from the measured concentrations of substrates and/or products may be significantly affected by the volume of biomass in the fermenting medium. Corrections proposed in this paper should be evaluated and, depending on their magnitude, considered in order to obtain more representative results.     

Effect of initial pH independent of operational pH on hydrogen fermentation of food waste

The effect of initial pH from 5.0 to 9.0 on H₂ fermentation of food waste was investigated. In this batch experiment, however, unlike previous studies for initial pH, operational pH was maintained at 5.0 by the addition of alkaline solution. Although the period for pH drop from the initial values to 5.0 was less than one-tenth of the entire fermentation, this short period significantly affected the H₂ production performance. At initial pH 6.0-9.0, successful H₂ yield of 1.3-1.9 mol H₂/mol hexose_added was achieved with a peak value at pH 8.0. The H₂ yield achieved at initial pH 8.0 was corresponded to the 8.13% of total energy content in the substrate. At initial pH 5.0, the smallest butyrate production, but the highest ethanol production was detected, indicating unfavorable conditions for H₂ production. There was no significant relationship between total required amount of alkaline solution and initial pH values.     

Production of the fungal exopolysaccharide pullulan by batch-wise and continuous fermentation

A mutant strain of the deuteromycete Aureobasidium pullulans deficient in melanin synthesis was used to investigate the production of the exopolysaccharide pullulan and biomass, respectively. Shake-flask experiments with different carbon sources showed significant differences in pullulan elaboration. Sucrose was most suitable for pullulan synthesis among the carbon sources examined. Fermentations were carried out both batch-wise and continuously in a stirred vessel fermentator. In batch fermentations about 45% of the glucose offered was converted into pullulan at maximum formation rates of 0.16 g/l per hour using standard medium. The yield of polysaccharide could be maintained at 45% in continuous fermentations. At a dilution rate of 0.05 l/h, the formation rate of polysaccharide increased up to 0.35 g/l per hour. Alterations in the nitrogen content of the feed significantly affected the consumption rate of glucose and the production rate of polysaccharide, but final concentrations of biomass were hardly affected. Correspondence to: R. Schuster