Dynamic modeling of an immobilized cell reactor

A mathematical model has been developed to describe the dynamic aerobic reaction occurring in a semibatch type of mixed flow reactor, containing cells immobilized in gel beads. This modeling is an extension of that developed in our previous study, for an immobilized cell reactor involving ethanol fermentation. In contrast to anaerobic reactions such as ethanol fermentation, (wherein the influent substrate concentration can be set at any desired level), aeration becomes necessary to provide additional substrate (oxygen) for most aerobic reactions occurring in immobilized cell reactors. Tobacco cell cultivation was chosen as a representative aerobic reaction, and the effect of aeration was assessed in terms of the volumetric coefficient of oxygen from gas to liquid phases.     

The Use of Microporous Divinyl Benzene Copolymer for Yeast Cell Immobilization and Ethanol Production in Packed-Bed Reactor

Microporous divinyl benzene copolymer (MDBP) was used for the first time as immobilization material for Saccharomyces cerevisiae ATCC 26602 cells in a bed reactor and ethanol production from glucose was studied as a model system. A very homogenous thick layer of yeast cells were seen from the scanning electron micrographs on the outer walls of biopolymer. The dried weight of the cells was found to be approximately 2 g per gram of cell supporting material. Hydrophobic nature of polymer is an important factor increasing cell adhesion on polymer pieces. The dynamic flow conditions through the biomaterial due to its microporous architecture prevented exopolysaccharide matrix formation around cells and continuous washing out of toxic metabolites and dead and degraded cells from the reactor provided less diffusional limitations and dynamic living environment to the cells. In order to see the ethanol production performance of immobilized yeast cells, a large initial concentration range of glucose between 6.7 and 300 g/l was studied at 1 ml/min in continuous packed-bed reactor. The inhibition effect of glucose with increasing initial concentration was observed at above 150 g/l, a relatively high substrate concentration. The continuous fluid flow around the microenvironment of the attached cells and mass transferring ability of cell immobilized on MDBP can help in decreasing the inhibition effect of ethanol accumulation and high substrate concentration in the vicinity of the cells.     

Nanosized reactors based on polyethyleneimines: from microheterogeneous systems to immobilized catalysts

Effective nanoreactors based on polyethyleneimines (PEIs) for the hydrolytic cleavage of O-alkyl O-p-nitrophenyl chloromethylphosphonates (alkyl = ethyl, hexyl) and di(p-nitrophenyl)phosphate were developed in conformity with the idea of modeling the polyfunctional catalytic mechanism of enzymes. A step-by-step modification of the single PEI solution by additives with their own catalytic activities (sodium dodecyl sulfate and lanthanum salt) gave rise to a marked improvement in the reaction efficiency. A 104-106-fold acceleration of the reaction compared to the aqueous basic hydrolysis of the substrates was achieved in the sodium dodecyl sulfate-polyethyleneimine-La(III) ternary system. This system can be considered to be metallomicelles immobilized on a hydrophilic polymer matrix. When the PEI immobilized on silica gel was used as a catalyst, the full completion of the reaction was achieved for 100 min under mild conditions, while the half-life of the reaction in a comparable homogeneous regime exceeds 100 h.     

京尼平交联低聚乙烯亚胺智能基因载体的制备与表征

使用京尼平与分子量为1800的超支化低聚乙烯亚胺在70％的乙醇溶液中反应,合成了具有荧光的交联型聚合物.利用核磁、凝胶渗透色谱、粒度仪、zeta电位仪和凝胶阻滞电泳对聚合物载体及其与DNA复合物颗粒进行了表征.研究表明,聚合物载体与DNA复合物颗粒粒径为120 ～150 nm,zeta电位为+20～25 mV,聚合物/...     

Ethanol Production Using Immobilized <Emphasis Type="Italic">Saccharomyces cerevisiae</Emphasis> in Lyophilized Cellulose Gel

A new lyophilization technique was used for immobilization of Saccharomyces cerevisiae cells in hydroxyethylcellulose (HEC) gels. The suitability of the lyophilized HEC gels to serve as immobilization matrices for the yeast cells was assessed by calculating the immobilization efficiency and the cell retention in three consecutive batches, each in duration of 72 h. Throughout the repeated batch fermentation, the immobilization efficiency was almost constant with an average value of 0.92 (12–216 h). The maximum value of cell retention was 0.24 g immobilized cells/g gel. Both parameters indicated that lyophilized gels are stable and capable of retaining the immobilized yeast cells. Showing the yeast cells propagation within the polymeric matrix, the scanning electron microscope images also confirmed that the lyophilization technique for immobilization of S. cerevisiae cells in the HEC gels was successful. The activity of the immobilized yeast cells was demonstrated by their capacity to convert glucose to ethanol. Ethanol yield of 0.40, 0.43 and 0.30 g ethanol/g glucose corresponding to 79%, 84% and 60% of the theoretical yield was attained in the first, second and third batches, respectively. The cell leakage was less than 10% of the average concentration of the immobilized cells.     

Continuous Production of Ethanol from Starch Using Glucoamylase and Yeast Co-Immobilized in Pectin Gel

This work presents a continuous simultaneous saccharification and fermentation (SSF) process to produce ethanol from starch using glucoamylase and Saccharomyces cerevisiae co-immobilized in pectin gel. The enzyme was immobilized on macroporous silica, after silanization and activation of the support with glutaraldehyde. The silica–enzyme derivative was co-immobilized with yeast in pectin gel. This biocatalyst was used to produce ethanol from liquefied manioc root flour syrup, in three fixed bed reactors. The initial reactor yeast load was 0.05 g wet yeast/ml of reactor (0.1 g wet yeast/g gel), used in all SSF experiments. The enzyme concentration in the reactor was defined by running SSF batch assays, using different amount of silica–enzyme derivative, co-immobilized with yeast in pectin gel. The chosen reactor enzyme concentration, 3.77 U/ml, allowed fermentation to be the rate-limiting step in the batch experiment. In this condition, using initial substrate concentration of 166.0 g/l of total reducing sugars (TRS), 1 ml gel/1 ml of medium, ethanol productivity of 8.3 g/l/h was achieved, for total conversion of starch to ethanol and 91% of the theoretical yield. In the continuous runs, feeding 163.0 g/l of TRS and using the same enzyme and yeast concentrations used in the batch run, ethanol productivity was 5.9 g ethanol/l/h, with 97% of substrate conversion and 81% of the ethanol theoretical yield. Diffusion effects in the extra-biocatalyst film seemed to be reduced when operating at superficial velocities above 3.7 × 10⁻⁴ cm/s.     

固定化青霉素酰化酶新型载体PEI/SiO2的制备及其特性

通过γ-氯丙基三甲氧基硅烷的媒介, 将聚乙烯亚胺(PEI)化学偶联在硅胶微粒表面, 制备了固定化青霉素酰化酶的新型复合载体PEI/SiO2, 最终制得了活性高且稳定性好的固定化青霉素酰化酶. 通过测定复合载体表面PEI的偶合量, 考察了各种反应条件对复合载体制备的影响规律; 通过红外光谱与电导滴定法测定, 对复合载体表面的化学结构与组成进行了表征; 为探索复合载体PEI/SiO2固定化酶的作用机理, 测定了复合载体在固定化酶前的ζ电位. 研究结果表明, 通过氯丙基硅烷偶联剂的媒介, 聚胺大分子PEI可以充分地被化学偶联在SiO2表面, 键合量可达到15%. 偶联反应的适宜条件: 反应温度90-94 ℃; 反应时间5h; PEI的质量浓度0.45-0.50 g/mL. 由于PEI分子链中含有大量氨基, 少量的共价键联与大量的物理吸附相结合, 既可使青霉素酰化酶被快速稳定地固定化, 又能很好地保持酶的构象, 使其具有较高的催化活性与活力回收率, 而且具有良好的连续操作稳定性, 重复使用15次, 固定化酶的活性可稳定地保持在初活性的87.5%水平上.     

Modeling of saccharide utilization in primary beer fermentation with yeasts immobilized in calcium alginate

Immobilized beer fermentation was studied using an industrial bottom-fermenting yeast strain Saccharomyces cerevisiae. The yeast cells were immobilized in 2.5% calcium alginate gel and used for brewing in a five-vessel cascade reactor. The fermentation was performed at 15°C at various flow rates. A nonstructured mathematical model was developed to simulate the performance of continuous primary fermentation of lager beer. The model was based on the following variables: maltose, maltotriose, glucose, fructose, ethanol, and cell concentration. Experimental values of these variables were determined in samples taken at regular intervals. For experimental data fitting a nonlinear regression was used. Substrate consumption was characterized by specific substrate consumption rate and saturation constant. The values of these two parameters were optimized for all four substrates. Inhibition effects of substrates and product were analyzed using various inhibition patterns. Only the inhibition effect of maltose on maltose consumption was clearly identified. A good-fitting relationship for maltose inhibition was found, and inhibition constants were calculated.     

Growth of yeast cells immobilized with porous swelling carriers produced by radiation polymerization

A new method of immobilization of yeast cells for the advantageous growth of cells inside and on the surface of a polymer carrier by physical adsorption is proposed. Porous and swellable polymer carriers were prepared by radiation-induced polymerization at low temperature. These polymer carriers were incubated with yeast cells at 30°C under aerobic conditions. Yeast cells were adsorbed on the surface of polymer carriers and subsequently infiltrated the polymer carriers by multiplication. The ethanol productivity of immobilized growing yeast cells thus obtained was thirteen times that of free yeast cells in a 1:1 volume of liquid medium to immobilized yeast cells.     

Development of continuous fermentation using immobilized yeast cells for wine cooler process development

The continuous wine fermentation process, which employs a newly designed tapered column type bioreactor and immobilized yeast cells (Montrachet 522), was studied and its fermentation performance was compared with batch and suspended cell continuous wine fermentation systems. It was found that a stable continuous culture fermentation process could be maintained for a period of 2–3 mo when the new bioreactor system packed with immobilized yeast cells was employed. The new bioreactor containing immobilized yeast cells performed significantly better than the suspended cell culture system or batch culture. The effluent wine from the continuous fermentor system contained 7.1% (v/v) ethanol and 0.18% (w/v) residual sugar at 0.01 h^-1 dilution rate. The new continuous bioreactor system also gave 17–34 times higher maximum ethanol productivity compared to the conventional batch wine fermentation. At a low dilution rate, 0.01^-1, as high as 92% sugar to ethanol yield was achieved. Based on the results obtained from this study, the possibility of developing a continuous wine cooler fermentation process was demonstrated. A two-stage continuous wine fermentation system may be designed and operated. The grape juice can be fed into the first-stage that is operated at about 0.2 h^-1 dilution rate and the effluent from the first-stage is fed into the second-stage continuous fermentor operated at about 0.01 h^-1 dilution rate. By doing so, a wine cooler can be produced continuously and efficiently, by employing the newly designed tapered column type bioreactor charged with the immobilized yeast cells.     

Continuous production of butanol with immobilized cells ofClostridium acetobutylicum

Spores ofClostridium acetobutylicum were immobilized in calcium alginate. An active gel preparation was obtained after outgrowth of the spores to vegetative cells within the gel matrix. A 100 mL column containing the immobilized cells was used for continuous production. At steady-state conditions the productivity of butanol was 67 g/L reactor volume/day.     

Comparison of Alcoholic Fermentation Performance of the Free and Immobilized Yeast on Water Hyacinth Stem Pieces in Medium with Different Glucose Contents

Ethanol fermentation with Saccharomyces cerevisiae cells was performed in medium with different glucose concentrations. As the glucose content augmented from 200 to 250 g/L, the growth of the immobilized cells did not change while that of the free cells was reduced. At higher glucose concentration (300, 350, and 400 g/L), the cell proliferation significantly decreased and the residual sugar level sharply augmented for both the immobilized and free yeast. The specific growth rate of the immobilized cells was 27–65 % higher than that of the free cells, and the final ethanol concentration in the immobilized yeast cultures was 9.7–18.5 % higher than that in the free yeast cultures. However, the immobilized yeast demonstrated similar or slightly lower ethanol yield in comparison with the free yeast. High fermentation rate of the immobilized yeast was associated with low unsaturation degree of fatty acids in cellular membrane. Adsorption of S. cerevisiae cells on water hyacinth stem pieces in the nutritional medium decreased the unsaturation degree of membrane lipid and the immobilized yeast always exhibited lower unsaturation degree of membrane lipid than the free yeast in ethanol fermentation.     

Fluorescence monitoring of immobilzed microorganisms in cultures

An experimental reactor system for monitoring the fluorescence of suspended and immobilized cells is described. The growth of S. cerevisiae was monitored during batch fermentations by fluorescence of the culture. Thus, it was possible to use this intracellular parameter to study the influence of immobilization on cells. The fermentations were done under aerobic conditions with suspended and immobilized cells. A comparison of these two systems showed that the rate of ethanol consumption was significantly slower for the cells immobilized in calcium alginate. This reduced rate of oxidative decomposition may be due to mass-transfer limitations of oxygen. Pulse experiments with different substrates (glucose and ethanol) were made to monitor the changes in cell metabolism. The reactor system presented is also suitable as a “toxin guard system”, because substances toxic to cells, such as 2,4-dinitrophenol, cause clearly visible changes in the fluorescence of the immobilized cells.     

Impact of High Temperature on Ethanol Fermentation by Kluyveromyces marxianus Immobilized on Banana Leaf Sheath Pieces

Ethanol fermentation was carried out with Kluyveromyces marxianus cells at various temperatures (30, 35, 40, and 45 °C). Fermentation performance of the immobilized yeast on banana leaf sheath pieces and the free yeast were evaluated and compared. Generally, ethanol production of the immobilized and free yeast was stable in a temperature range of 30–40 °C. Temperature of 45 °C restricted yeast growth and lengthened the fermentation. The immobilized yeast demonstrated faster sugar assimilation and higher ethanol level in the fermentation broth in comparison with the free yeast at all fermentation temperatures. Change in fatty acid level in cellular membrane was determined to clarify the response of the free and immobilized yeast to thermal stress. The free cells of K. marxianus responded to temperature increase by increasing saturated fatty acid (C16:0 and C18:0) level and by decreasing unsaturated fatty acid (C18:1 and C18:2) level in cellular membrane. For fermentation at 40 °C with immobilized cells of K. marxianus, however, the changes were not observed in both saturated fatty acid (C16:0) and unsaturated fatty acid (C18:1 and C18:2) level.     

A study of ethanol production of yeast cells immobolized with polymer carrier produced by radiation polymerization

Polymer carriers, poly(hydroxyethyl acrylate(HEA)-methoxy polyethylene glycol methylacrylate (M-23G)) and poly (hydroxyethyl accrylate(HEA)-glycidyl methlacrylate(GMA)) using for immobilization of yeast cells were prepared by radiation polymerization at low temperature. Yeast cells were immobilized through adhesion and multiplication of yeast cells themselves. The ethanol productivity of immobilized yeast cells with these carriers was related to the monomer composition of polymers and the optimum monomer composition was 20% : 10% in poly(HEA-M-23G) and 17%: 6% in poly(HEA-GMA). In this case, the ethanol productivity of immobilized yeast cells was 29mg/ml/h which was about 4 times that of cells in free system. The relationship between the activity of immobilized yeast cells and the water content of polymer carrier were also discussed.     

Modeling and analysis of high‐impact poly(propylene) production processes, 1. Effect of chemical poisoning on particle size distribution and gel formation

This work presents a simple model for a two‐stage process of high impact poly(propylene) (HIPP) production. The model predicts the bivariate distribution of particle size and polymer composition. It takes into account the effect of chemical poisoning on gel particle formation. The result shows that poisoning the solid catalyst is not an effective method for gel reduction. A better approach is to saturate the polymer particles with a co‐catalyst in reactor 1 and poison the co‐catalyst in reactor 2. It is also shown that the residence time distribution (RTD) of reactor 1 has a strong effect on the gel particle formation. A continuous reactor with narrow RTD is advantageous for gel reduction. The model provides some guidance for the analysis and design of the HIPP production process.     

Ethanol production by a flocculant yeast strain in a CSTR type fermentor with cell recycling

Tests were performed in a continuous stirred tank reactor (CSTR), with and without cell recycling, to produce ethanol. The reactor without cell recycling produced the kinetic model of ethanol production, whereas the reactor with cell recycling allowed for a study of process stability. The Levenspiel kinetic model was adopted; however, in the case of fermentation with cell recycling, the coefficient of cell death was added. It was observed that cellular viability varied greatly throughout the fermenting process and that microaeration is of fundamental importance in maintaining the stability of the process.     

Cell immobilization and xylitol production using sugarcane bagasse as raw material

Sugarcane bagasse pretreated by three different procedures (with 2% [v/v] polyethyleneimine (PEI), with 2% [w/v] NaOH, or with a sequence of NaOH and PEI) was used as cell immobilization carrier for xylitol production byCandida guilliermondii yeast. Fermentations using these pretreated carriers were performed in semidefined medium and in a hydrolysate medium produced from sugarcane bagasse hemicellulose. Sugarcane bagasse pretreated with NaOH was the best carrier obtained with respect to immobilization efficiency, because it was able to immobilize a major quantity of cells (0.30 g of cells/g of bagasse). Fermentation in semidefined medium using the NaOH-pretreated carrier attained a high efficiency of xylose-to-xylitol bioconversion (96% of the theoretical value). From hydrolysate medium, the bioconversion efficiency was lower (63%), probably owing to the presence of other substances in the medium that caused an inadequate mass transfer to the cells. In this fermentation medium, better results with relation to xylitol production were obtained by using PEI-pretreated carrier (xylose-to-xylitol bioconversion of 81% of the theoretical and volumetric productivity of 0.43 g/[L·h]). The results showed that sugarcane bagasse is a low-cost material with great potential for use as cell immobilization carrier in the fermentative process for xylitol production.     

Characteristics of an Immobilized Yeast Cell System Using Very High Gravity for the Fermentation of Ethanol

The characteristics of ethanol production by immobilized yeast cells were investigated for both repeated batch fermentation and continuous fermentation. With an initial sugar concentration of 280?g/L during the repeated batch fermentation, more than 98% of total sugar was consumed in 65?h with an average ethanol concentration and ethanol yield of 130.12?g/L and 0.477?g ethanol/g consumed sugar, respectively. The immobilized yeast cell system was reliable for at least 10 batches and for a period of 28?days without accompanying the regeneration of Saccharomyces cerevisiae inside the carriers. The multistage continuous fermentation was carried out in a five-stage column bioreactor with a total working volume of 3.75?L. The bioreactor was operated for 26?days at a dilution rate of 0.015?h^?1. The ethanol concentration of the effluent reached 130.77?g/L ethanol while an average 8.18?g/L residual sugar remained. Due to the high osmotic pressure and toxic ethanol, considerable yeast cells died without regeneration, especially in the last two stages, which led to the breakdown of the whole system of multistage continuous fermentation.     

Radiation synthesis of acrylic acid/polyethyleneimine interpenetrating polymer networks (IPNs) hydrogels and its application as a carrier of atorvastatin drug for controlling cholesterol

Gamma irradiation was used to form interpenetrating polymer networks structure (IPNs) hydrogels based on different ratios of acrylic acid monomer (AAc) and polyethyleneimine (PEI). The property-behavior was characterized by IR spectroscopy, gel content, thermogravimetric analysis (TGA) and swelling in water at room temperature and different pH values. The AAc/PEI hydrogels were used as a carrier for atorvastatin drug, in which the uptake-release character was studied. The results showed that the gel content of AAc/PEI hydrogels decreased greatly with increasing the ratio of PEI in the initial feeding solution. The AAc/PEI hydrogels displayed pH-sensitive character. The drug uptake-release study indicated that AAc/PEI hydrogels possessed controlled release behavior and that the release process depends on pH. In this respect, the release of atorvastatin drug was significant in acidic medium.