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At least 19 recordsLinked to original sources

[Study on immobilized cells for producing alpha-amylase by using polyving alcohol as the carrier(II): The effect of fermentating conditions on the ability producing alpha-amylase of the cells immobilized with polyving alcohol as the corrier and continuous fermentation of the immobilized cells in CSTR].

The effects of fermentating conditions on the ability of immobilized cells with PVA as carrier for producing alpha-amylase were studied. The continuous fermentation with the immobilized cells were tested in continuous flow stirred tank reactor (CSTR). The results showed that the adaptability of the immobilized Bacillus substilis to pH increased after immobilization. In CSTR, the immobilized cells can be fermentated continuously for 360 hrs and the activity of alpha-amylase can be kept on the level of about 170 u/ml.

Bacillus↗

Whole-cell immobilization using cell surface-exposed cellulose-binding domain.

Specific adhesion of Eshcherichia coli with surface-exposed cellulose-binding domain (CBD) to cellulosic materials was investigated. Whole-cell immobilization was very specific, forming essentially a monolayer of cells onto the different supports. Cells with surface-exposed CBD bound specifically and tightly to cellulose supports at a wide range of pH. In contrast to CBD, which shows the highest binding to cellulose at 4 degrees C, highest cell loading was observed at 37 degrees C. The extent of immobilization was dependent on the amount of surface-exposed CBD. Cells binding increased with increasing amount of CBD until binding was saturated. Even induction of very low level of CBD (0.05 mM IPTG) was sufficient to provide specific and tight binding to cellulose support. Because optimal binding can be obtained under physiological conditions such as pH 7 and 37 degrees C, the results demonstrate the general utility of surface-exposed CBD as an efficient means of whole-cell immobilization.

Bacterial Outer Membrane Proteins↗

[Nisin formation by immobilized cells of the lactic acid bacterium, Streptococcus lactis].

The problem of microbial cell immobilization at present attracts the ever increasing attention of the scientists, since such organisms may be the source of various enzymes. Production of nizin by the immobilized cells of Str. lactis was studied. It was found that the cells of Str. lactis incorporated into polyacrylamide gel produced nizit on definite media. Still, the amount of the antibiotic was 2-3 times lower than in case of using free cells. The effect of a number of factors on the process of immobilization was studied and the influence of some factors, such as temperature, pH, aeration on nizin synthesis by the immobilized cells of the streptococcus was elucidated. Optimal conditions for nizin biosynthesis by the immobilized cells of Str. lactis were developed.

Acrylamides↗

Study on the mechanism of Bioelectric Recognition Assay: evidence for immobilized cell membrane interactions with viral fragments.

The Bioelectric Recognition Assay (BERA) is a whole-cell based biosensing system that detects the electric response of cultured cells, suspended in a gel matrix, to various ligands, which bind to the cell and/or affect its physiology. Previous studies have demonstrated the potential application of this method for rapid, inexpensive detection of viruses in a crude sample. However, the understanding, so far, of the fundamental processes that take place during cell-virus interactions within the probe has been rather limited. In the present study, we combined electrophysiological and fluorescence microscopical assays, so that we can prove that animal and plant cells immobilized in BERA sensors respond to different viruses primarily by changing their membrane potential. The response of immobilized cells against different viruses did not depend on the virus ability to penetrate the cell, but was modified after binding each virus to a virus-specific antibody or removal of its coat protein after treatment with a protease. Consequently, we were able to assay the presence of a virus in its complete form or fragments thereof. Combination of immunological recognition with the electrophysiological response of immobilized cells allows for a considerable increase of the specificity of the BERA biosensory assay. In addition, rather than simply detect the presence of a protein or genomic sequence, the method can help gain information on the bioactivity of a virus.

Animals↗

The role of cell immobilization in fermentation technology.

The distinction between immobilized cell fermentation and immobilized cell biocatalysis is seldom made, though they are conceptually quite different. Unlike immobilized enzyme systems, immobilized viable cells can be used to carry out conventional fermentations. Microbial cells which would otherwise be freely dispersed (in almost colloidal suspension) within the fermentation environment can be encouraged to become attached in some way to a support (carrier), thus producing a discrete particulate solid phase. Such immobilization offers several potential advantages of a process engineering nature to the fermentation system. These include ease of handling and of cell separation, and lowering of bulk viscosity, as well as the obvious potential benefits of increased cell concentration.

Biotechnology↗

[Degradation of 2,4-dinitrophenol by free and immobilized cells of Rhodococcus erythropolis HL PM-1].

Degradation of 2,4-dinitrophenol (2,4-DNP) by the cells of Rhodococcus erythropolis HL PM-1 was studied. The enzymes involved in 2,4-DNP degradation were inducible, and their resynthesis took place during the process. Cell immobilization by embedding into agar gels decreased the degrader activity. Maximum rates of 2,4-DNP degradation by free and immobilized cells were 10.0 and 5.4 nmol/min per mg cells, respectively. The concentration dependence of 2,4-DNP degradation was typical of substrate inhibition kinetics. The immobilized cells were used in a model reactor designed for 2,4-DNP biodegradation. Its maximum capacity was 0.45 nmol/min per mg cells at a volumetric flow rate of 20 h-1. The reactor operated for 14 days without losing capacity; its half-lifetime equaled 16 days.

2,4-Dinitrophenol↗

Investigations on neomycin production with immobilized cells of Streptomyces marinensis NUV-5 in calcium alginate matrix.

The purpose of this investigation was to study the effect of Streptomyces marinensis NUV-5 cells immobilized in calcium alginate for the production of neomycin. The effect of various parameters, such as the effect of alginate concentration (1%, 2%, 3%, 4%, and 5% wt/vol), the effect of cation (CaCl2, BaCl2, and SrCl2), the concentration of cation (0.01M, 0.125M, 0.25M, 0.375M, and 0.5M), the curing times (1, 6, 11, 16, and 21 hours), and the diameter of the bead (1.48, 2.16, 3.24, 4.46, and 5.44 mm), on neomycin production and bead stability were studied. The effect of maltose (4%, 3%, 2%, and 1% wt/vol) and sodium glutamate (0.6%, 0.3%, 0.15%, and 0.075% wt/vol) concentration on neomycin production was also studied. Better neomycin production was achieved with optimized parameters, such as alginate at 2% wt/vol, 0.25M CaCl2, 1-hour curing time, and 3.24 mm bead diameter. Effective neomycin production was achieved with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate concentration. The repeated batch fermentations were conducted (every 96 hours) using the optimized alginate beads, employing the production medium with 3% wt/vol maltose and 0.6% wt/vol sodium glutamate along with mineral salts solution. The increase in antibiotic production was observed up to the 5th cycle, and later gradual decrease in antibiotic production was observed. Comparison of the total antibiotic production with free cells and immobilized cells was also done. An enhanced antibiotic productivity of 32% was achieved with immobilized cells over the conventional free-cell fermentation, while 108% more productivity was achieved over the washed free-cell fermentation. From these results it is concluded that the immobilized cells of S marinensis NUV-5 in calcium alginate are more efficient for the production of neomycin with repeated batch fermentation.

Alginates↗

Red wine making by immobilized cells and influence on volatile composition.

Red wine making using yeast cells immobilized in two types of raisin berries, at various temperatures (6-30 degrees C), was studied. A modification of the batch bioreactor was used to separate the grape skins used for color extraction from the biocatalyst and the fermenting grape must. The evaluation of the immobilized biocatalysts was made on terms of productivity and organoleptic quality, including color intensity and formation of volatiles. The immobilized cells were found capable of low-temperature wine making, producing red wines containing more than 11% v/v alcohol in 8 days at 6 degrees C. The quality of wines was examined by gas chromatography (GC) and GC-MS analysis and sensory evaluation. Higher alcohol concentrations were decreased, and ethyl acetate concentrations increased by the drop of temperature. Many esters, alcohols, carbonyls, and miscellaneous compounds were identified in wines produced by immobilized cells, revealing no significant qualitative differences as compared to wines produced by free cells. The sensory evaluation showed that the best red wine was produced at 6 degrees C.

Bioreactors↗

Residence time distribution in a packed bed bioreactor containing porous glass particles: influence of the presence of immobilized cells.

An experimental investigation of the liquid phase residence time distribution (RTD) in a packed bed bioreactor containing porous glass particles is presented. For Re < 1, intraparticle forced convection is negligible and only diffusion, characterized by an effective diffusion coefficient, must be considered to describe the mass transfer process between the extraparticle and the intraparticle fluid phase. For Re > 1, the mass transfer rate becomes dependent on the liquid flow rate, indicating the existence of intraparticle convection. A model including axially dispersed flow for the external fluid phase and an 'apparent' effective diffusivity that combines diffusion and convection, predicts experimental RTD data satisfactorily. Yeast cells immobilized inside the porous glass beads did not affect the mass transfer rate at low biomass loading. At high biomass loading (0.02 g yeast cells g-1 carrier), the mass transfer rate between the extraparticle and intraparticle fluid phase was significantly decreased. Comparison of the RTD data from experiments performed in the presence and absence of cells in the external fluid phase revealed that the mass transfer rate is influenced by the cells immobilized inside the porous particles and not by the cells present in the external fluid phase.

Biotechnology↗

Methods for the estimation of the number and quality of animal cells immobilized in carbohydrate gels.

Rapid and reliable methods for the determination of survival, proliferation, and metabolic activity of immobilized cells in gels are described. The first method is based on an MTT assay that measures qualitatively and quantitatively the metabolic activity of the cells. The second method determines cell number by measuring the amount of DNA available for Feulgen staining. In the third method, two fluorescent dyes are used to differentially stain viable and dead cells. The fourth method involves the use of glutaraldehyde to protect the cells when melting the gel to facilitate hemocytometric count. The presented techniques should help to test the efficiency of the immobilization procedures and to monitor the growth and survival of immobilized cells.

Animals↗

Food additive lactic acid production by immobilized cells of Lactobacillus brevis on delignified cellulosic material.

Improvements in yield and productivity in lactic acid fermentation by Lactobaccilus brevis cells immobilized on delignified cellulosic (DC) material are reported. The system proved to be more efficient in comparison with the work reported by other workers. Yields of 80 and 100% conversion using glucose were obtained at 30 degrees C in 1 day of fermentation time. Lactic acid fermentation using whey as substrate was obtained at 30 degrees C in 1-1.5 days, resulting in 70% yield, whereas the remaining lactose in whey was converted to alcohol byproduct, leading to a 90% lactose exploitation and 100% conversion. Cell immobilization of L. brevis on DC material was proved by its reuses in repeated batch fermentations and through electron microscopy. A series of 10 repeated batch fermentations without any loss in cell activity showed a tendency for high operational stability. The presence of DC material resulted in a drastic drop of the fermentation time from 48 to 13 h.

Cellulose↗

Enhancement and stabilization of the production of glucoamylase by immobilized cells of Aureobasidium pullulans in a fluidized-bed reactor.

Glucoamylase production by Aureobasidium pullulans A-124 was compared in free-living cells, cells immobilized in calcium alginate gel beads aerated on a rotary shaker (agitation rate 150 rpm), and immobilized cells aerated in an air bubble column reactor. Fermentation conditions in the bioreactor were established for bead concentration, substrate (starch) concentration, calcium chloride addition to the fermentation medium, and rate of aeration. Production of glucoamylase was optimized at approximately 1.5 units of enzyme activity/ml medium in the bioreactor under the following conditions: aeration rate, 2.0 vol air per working volume of the bioreactor (280 ml) per minute; gel bead concentration, 30% of the working volume; substrate (starch) concentration, at 0.3% (w/v); addition of calcium chloride to the medium at a final concentration of 0.01 M. Productivity levels were stabilized through the equivalent of ten batches of medium with the original inoculum of immobilized beads.

Alginates↗

A structured model for immobilized cell kinetics.

An intrinsic, structured model has been formulated to describe the kinetics of viable (living) cells immobilized within porous supports. Predictions of steady-state internal biomass concentration distributions, biocatalyst substrate profiles, and immobilized cell growth and leakage from the support are in qualitative agreement with the literature. Simulation studies indicate that carrier pore structure is a particularly important design variable to be optimized.

Catalysis↗

Production of alkaline protease with immobilized cells of Bacillus subtilis PE-11 in various matrices by entrapment technique.

The purpose of this investigation was to study the effect of Bacillus subtilis PE-11 cells immobilized in various matrices, such as calcium alginate, k-Carrageenan, ployacrylamide, agar-agar, and gelatin, for the production of alkaline protease. Calcium alginate was found to be an effective and suitable matrix for higher alkaline protease productivity compared to the other matrices studied. All the matrices were selected for repeated batch fermentation. The average specific volumetric productivity with calcium alginate was 15.11 U/mL/hour, which was 79.03% higher production over the conventional free-cell fermentation. Similarly, the specific volumetric productivity by repeated batch fermentation was 13.68 U/mL/hour with k-Carrageenan, 12.44 U/mL/hour with agar-agar, 11.71 U/mL/hour with polyacrylamide, and 10.32 U/mL/hour with gelatin. In the repeated batch fermentations of the shake flasks, an optimum level of enzyme was maintained for 9 days using calcium alginate immobilized cells. From the results, it is concluded that the immobilized cells of B subtilis PE-11 in calcium alginate are more efficient for the production of alkaline protease with repeated batch fermentation. The alginate immobilized cells of B subtilis PE-11 can be proposed as an effective biocatalyst for repeated usage for maximum production of alkaline protease.

Alginates↗

A diffusion model and optimal cell loading for immobilized cell biocatalysts.

A diffusion model based on the random pore model is derived for immobilized cell biocatalysts and verified with 19 sets of experimental diffusion data. The predicted effective diffusivity relative to that for the support matrix reflects a quadratic dependence on the cell loading and contains a single parameter that depends on the intracellular diffusivity and the chemical partitioning coefficient. The model is used to predict optimal cell loadings that maximize the total reaction rate in an immobilized cell biocatalyst. A rule of thumb based on the diffusion model is obtained to the effect that the cell loading should be at least (1/3) for single reactions regardless of the kinetics and diffusional resistances. A means of calculating improved lower bounds is provided for cases where the cellular diffusional resistance is known but the kinetics are not. The optimal cell loadings for reversible first-order and for Michaelis-Menten kinetics are presented and demonstrated to be within the range of conditions of practical interest.

Biotechnology↗

Cell growth on immobilized cell growth factor: 5. Interaction of immobilized transferrin with fibroblast cells.

Transferrin was immobilized on poly(methyl methacrylate) membrane and the interaction of the immobilized transferrin with mouse fibroblast cells STO was investigated. The immobilized transferrin was stable without decomposition over 48 h and showed higher activities of cell growth acceleration than free transferrin and transported ferric ion as effectively as free transferrin. The specific interaction of the immobilized transferrin with fibroblast cells was confirmed by experiment using the antibody. It was considered that cell growth acceleration and ferric ion transportation occur by different mechanisms.

Animals↗

Bioconversion of nitriles by Candida guilliermondii CCT 7207 cells immobilized in barium alginate.

Nitrile degradation by Candida guilliermondii CCT 7207 using free and immobilized cell systems was compared. Different specific growth rates were observed for immobilized (mumax=0.021 h(-1)) and the free cells (mumax=0.029 h(-1)). The maximum specific rate of acetic acid formation was 0.387 h(-1) and 0.266 h(-1) for free and immobilized cells, respectively. Cell adhesion to the support materials was confirmed by scanning electron microscopy. When immobilized, the yeast was able to use high nitrile and amide concentrations (aliphatic and aromatic) as nitrogen sources. The results suggest that C. guilliermondii CCT 7207 presents a physiological pattern potentially useful for the bioremediation of polluted environments or for the bioproduction of amides and organic acid of high commercial value.

Alginates↗

Cell immobilization using self-assembled synthetic oligopeptide and its application to biological toxicity detection using surface plasmon resonance.

The immobilized cell using self-assembled synthetic oligopeptide was applied to the biological toxicity detection of environmental pollutant. Thin films based on cysteine-terminated synthetic oligopeptides were fabricated for the immobilization of Escherichia coli O157:H7 on gold (Au) substrate. Layer formation and immobilization of E. coli O157:H7 were investigated with surface plasmon resonance (SPR) and atomic force microscopy (AFM). Experimental results showed that the thin film of cysteine-terminated synthetic oligopeptide was successfully fabricated and it could be applied for the immobilization of E. coli O157:H7. The attached living cell was exposed to toxic chemical such as phenol, which induced the change of SPR angle. As the exposed concentration of phenol was increased, the change of plasmon resonance angle was increased, which indicates the decrease of cell viability. The detection limit based on SPR was determined as 5 ppm. The proposed cell immobilization method using self-assembly technique can be applied to construct the cell microarray for the diagnosis, drug detection, and on-site monitoring.

Biological Assay↗