Characterization of a pilot plant airlift tower loop bioreactor: II. Evaluation of global mixing properties of the gas phase during yeast cultivation

Saccharomyces cerevisiae was cultivated in a 4-m(3) pilot plant airlift tower loop reactor with a draft tube in batch and continuous operations and for comparison in a laboratory airlift tower loop reactor of 0.08 m(3) volume. The reactors were characterized during and after the cultivation by measuring the distributions of the residence times of the gas phase with pseudostochastic tracer signals and mass spectrometer and by evaluating the mixing in the liquid phase with a pulse-shaped volatile tracer signal and mass spectrometer as a detector. The mean residence times and the intensities of the axial mixing in the riser and downcomer, the circulation times of the gas phase, and the fraction of the recirculated gas phase were evaluated and compared.     

Observations Relating to the Efficiency of Mixing in Rapid Reaction Flow Devices

The principle of dimensional analysis of liquid flow has been applied to the problem of rapid mixing in flow apparatus. A model of the Hartridge-Roughton mixing chamber and observation tube has been scaled approximately 1/1.000 so that the times after mixing are approximately 6 s, the flow velocities are the order of 2 cm/s, and the distance from mixing to observation is 120 mm. Visual observation is employed to observe the end point of the mixing of acid-base with a color indicator, thymol blue. Quantitative estimates of the effect of the number of jets, the effect of screens placed in the flow stream, and the angle of jets, one with respect to another, lead to quantitative evaluations of these parameters for the scale model. The extent to which these parameters apply to the full-scale apparatus at faster flow velocities suggest that the general principles employed in the scale model are valid, although more emphasis is placed upon turbulence generation at the low flow velocities employed in this experiment than at the faster flow velocities employed in the full-scale apparatus.     

The intercellular interactions in the interphase death of irradiated thymocytes. A study of the nature of the mediator of the interaction of irradiated thymocytes]

A study was made of the influence of cytosol and conditioned medium, obtained at different times of incubation of irradiated thymocytes, on native thymocytes, as well as of the influence of mixing the suspension and changing the medium and the enzyme inhibitors, involved in metabolism of arachidonic acid, on the radiation injury to thymocytes. Cytosol of thymocytes, incubated for 60 min after irradiation, was shown to exert an insignificant toxic effect on native thymocytes. The conditioned medium of irradiated thymocytes is virtually ineffective. The mixing of the suspension upon incubation and the changing of the medium reduced the damage to exposed thymocytes while inhibitors of phospholipase A2 and lipoxygenase prevented completely the radiation-induced pyknosis of nuclei, whereas a inhibitor of cyclooxygenase did not influence it. It is inferred from the data obtained that unstable lipoxygenase products may serve as mediators of intercellular interactions of exposed thymocytes.     

Mixing in liquid-impelled loop reactors

The liquid-impelled loop reactor is a new column-type bioreactor. The design of this device is based on the principle of the air-lift loop reactor. In the external-loop configuration used in this work, descending perfluorochemical drops bring about circulation of the continuous aqueous phase. Mixing of this continuous phase is characterized per section of the rector. Axial-dispersion coefficients for the tube with two-phase flow are determined and correlated with the energy dissipation in the tube. Comparisons with similar systems such as bubble columns and air-lift loop reactors are made. Overall mixing parameters are derived and used for calculation of the number of circulatins needed to achieve a certain degree of mixing. The hydrodynamic model from previous work is tested for the reactor configurations of this work. It can be useful to calculate circulation times.     

Macromixing hydrodynamic study in draft-tube airlift reactors using electrical resistance tomography

The present study summarizes results of mixing characteristics in a draft tube airlift bioreactor using ERT. This technique offers the possibility for noninvasive and nonintrusive visualization of flow fields in the bioreactor and has rarely been utilized previously to analyze operating parameters and mixing characteristics in this type of bioreactors. Several operating parameters and geometric characteristics were examined. In general, results showed that the increase in superficial gas velocity corresponds to an increase in energy applied and thus, to a decrease in mixing time. This generally corresponded to an increase in liquid circulation velocity and shear rate values. Bottom clearances and draft tube diameters affected flow resistance and frictional losses. The influence of sparger configurations on mixing time and liquid circulation velocity was significant due to their effect on gas distribution. However, the effect of sparger configuration on shear rate was not significant, with 20% reduction in shear rates using the cross-shaped sparger. Fluid viscosity showed a marked influence on both mixing times and circulation velocity especially in the coalescing media of sugar and xanthan gum (XG) solutions. Results from this work will help to develop a clear pattern for operation and mixing that can help to improve several industrial processes, especially the ones related to emerging fields of technology such as the biotechnology industry.     

Loop-mediated isothermal amplification method for rapid detection of the toxic dinoflagellate Alexandrium, which causes algal blooms and poisoning of shellfish

The marine dinoflagellate genus Alexandrium includes a number of species that produce potent neurotoxins responsible for paralytic shellfish poisoning, which in humans may cause muscular paralysis, neurological symptoms and, in extreme cases, death. Because of the genetic diversity of different genera and species, molecular tools may help to detect the presence of target microorganisms in marine field samples. Here we employed a loop-mediated isothermal amplification (LAMP) method for the rapid and simple detection of toxic Alexandrium species. A set of four primers were designed based upon the conserved region of the 5.8S rRNA gene of members of the genus Alexandrium . Using this detection system, toxic Alexandrium genes were amplified and visualized as a ladder-like pattern of bands on agarose gels under isothermal condition within 60 min. The LAMP amplicons were also directly visualized by eye in the reaction tube by the addition of SYBR Green I. This LAMP assay was 10-fold more sensitive than a conventional PCR method with a detection limit of 5 cells per tube when targeting DNA from Alexandrium minutum . The LAMP assay reported here indicates the potential usefulness of the technique as a valuable simple, rapid alternative procedure for the detection of target toxic Alexandrium species during coastal water monitoring.     

Continuous refolding of lysozyme with fed-batch addition of denatured protein solution

A continuous refolding method with addition of denatured protein solution in a fed-batch manner through a ceramic membrane tube was developed. Denatured and fully reduced lysozyme was continuously refolded with high refolding efficiencies. In this method, a denatured lysozyme solution was gradually added from the outer surface of the membrane tube into a refolding buffer flowing continuously inside the tube under controlled mixing conditions. The refolding efficiencies of lysozyme in this continuous refolding were higher than those in a batch dilution method. This method has applicability to large-scale downstream processes and can attain a high efficiency and protein concentration in refolding. Refolded proteins can be supplied continuously following purification steps.     

A continuous-flow, rapid-mixing, photolabeling technique applied to the acetylcholine receptor

A continuous-flow technique is described in which a photoaffinity label, membrane rich in acetylcholine receptor, and various effectors are rapidly mixed, passed through a delay tube, through a tube in which they are irradiated, and are collected in a tube containing quencher. Delay times as short as 20 ms between mixing and photolysis are achievable. Because the flow is continuous, milliliter volumes of membrane can be labeled in a single run, which is convenient for the analysis of both the functional effects and sites of photolabeling. Using this technique, we have found that receptor in its transitory, active state, in which the channel is open, is more susceptible to photolabeling by the noncompetitive inhibitor analog [3H] quinacrine azide than is receptor in either its resting or desensitized states, in which the channel is closed. This technique should prove generally useful for the photolabeling of transient conformational states of macromolecules.     

Mixing time and oxygen transfer characteristics of double draft tube airlift fermentor

Despite the increasing importance of airlift fermentors, very little published information is available on how the geometric configurations of the draft tubes and the air-sparging system affect the mixing and oxygen transfer characteristics of the fermentor. A 14-L air-lift fermentor was designed and build with a fixed liquid height to diameter ratio of 1.5 utilizing four equally spaced air jets at the bottom. Two jet orifice sizes were used, 1.27 and 3.81 mm i.d., and for each jet size the following four geometric configurations were used: Single inner concentric draft tube, single outer concentric draft tube, two concentric draft tubes, and no draft tubes where the fermentor was operated as a shallow bubble column. It was found that the presence of draft tubes stabilized liquid circulation patterns and gave systemically higher mixing times than those obtained in the absence of draft tubes. In addition, the double draft tube geometry resulted in higher mixing times than the single draft tubes. For the power unit volume range 20 to about 250 W/m³ the larger 3.81-mm orifices gave systemically higher k_L a values than the smaller 1.27-mm i.d. orifices. At 200 W/m³ the use of a single outer draft tube with the 3.81-mm orifices resulted in 94% increase in k_L a values over that obtained with no draft tubes. However, the effect of draft tube geometry on k_L a values when the 1.27-mm orifices were used was not significant. The air bubble formation characteristics at the jet orifices were found to be different, which reflected the differences observed in mass transfer and mixing characteristics. The power economy for oxygen transfer was found to be depend strongly on the orifice size and less on the geometric configuration of draft tubes.     

Hydrodynamic performance of a three-phase airlift bioreactor with an enlarged degassing zone

The hydrodynamics of biotechnological processes is complex. So far, few studies were made with bioreactors of the airlift type with an enlarged degassing zone.In this work, the influence of solids loading, solids specific gravity and draught tube dimensions on mixing and circulation times and critical air flow rate for an internal loop airlift bioreactor with an enlarged sedimentation/degassing zone is studied.The results indicate that the critical air flow rate as well as the mixing time increase with an increase in solids loading in the bioreactor. Circulation time presents a maximum for a solids load between 5 and 10% (v/v). It is also shown that small variations in solids specific gravity, for values close to that of the liquid, have a significant influence on the critical air flow rate and on the mixing time.An optimal (minimal) value for the circulation time and for the critical air flow rate was obtained for a riser to down comer diameter ratio of 0.46. The minimum mixing time was obtained for a riser to down comer height ratio of 0.80.This work was supported by J.N.I.C.T. (Junta Nacional de Investigação Cientifica e Tecnológica).     

Pressure change and gas mixing induced by oscillations in a closed system

In an attempt to delineate some mechanical behaviors found in branching airways, pressure transmission, gas motion, and mixing were studied during high-frequency oscillation (HFO) in an idealized system consisting of a large straight tube and a rigid sphere linked together by a small straight tube. Depending on the frequency f, and on the unsteadiness dimensionless parameter alpha, pressure amplitude in the large tube is either strongly attenuated or amplified in the sphere. This finding may provide a theoretical basis for the pressure resonance phenomenon observed in the lung by previous investigators. Gas compression in the closed volume causes convective mixing throughout the system. The measured dispersion was found to be proportional to f(VT/A)2, in agreement with a recent report. However, bulk convective mixing was sufficient to explain the dispersion for oscillatory volumes (VT) as small as 80 percent of the small tube volume, as has been previously suggested.     

Two- and three-phase mixing in a concentric draft tube gas-lift fermentor

Two- and three-phase mixing studies were carried out in a 44-L concentric draft tube gas-lift fermentor. It was proposed to use the fermentor for the production of solvents using immobilized bacteria. Bubble size, gas holdup, liquid velocities, circulation, and mixing times were determined for various superficial gas velocities in distilled water, starch, carboxymethyl cellulose, and ethanol solutions. The observed trends for two phase mixing were similar to other studies but the results were found to be more sensitive to liquid properties. This was possibly due to the large value of downcomer to riser area used in this study. Mixing in three phases highlighted the difficulty in predicting the effect of adding solids to the gas-liquid system. Results showed that the gas-lift fermentor was ideally suited to dealing with three phases but more work is necessary before accurate models can be developed to account for the effect of solids.     

Oscillatory flow and gas transport through a symmetrical bifurcation

Axial gas transport due to the interaction between radial mixing and radially nonuniform axial velocities is responsible for gas transport in thick airways during High-frequency oscillatory ventilation (HFO). Because the airways can be characterized by a bifurcating tube network, the secondary flow in the curved portion of a bifurcating tube contributes to cross-stream mixing. In this study the oscillatory flow and concentration fields through a single symmetrical airway bifurcating tube model were numerically analyzed by solving three-dimensional Navier-Stokes and mass concentration equations with the SIMPLER algorithm. The simulation conditions were for a Womersley number, alpha = 9.1 and Reynolds numbers in the parent tube between 200 and 1000, corresponding to Dn2/alpha 4 in the curved portion between 2 and 80, where Dn is Dean number. For comparison with the results from the bifurcating tube, we calculated the velocity and concentration fields for fully developed oscillatory flow through a curved tube with a curvature rate of 1/10, which is identical to the curved portion of the bifurcating tube. For Dn2/alpha 4 < or = 10 in the curved portion of the bifurcating tube, the flow divider and area changes dominate the axial gas transport, because the effective diffusivity is greater than in either a straight or curved tube, in spite of low secondary velocities. However, for Dn2/alpha 4 > or = 20, the gas transport characteristics in a bifurcation are similar to a curved tube because of the significant effect of secondary flow.     

Sediment toxicity in the Kattegat and Skagerrak

Sediments were sampled from 62 sites in the Kattegat and Skagerrak, which are located between the Baltic and the North Sea in the Western Atlantic, during autumn 1989 and spring 1990. From each site 5 to 6 samples were taken wit ha box-corer. After mixing to composite samples on board, transport and storage (at 4 °C for 2 to 4 weeks), the samples were tested for toxicity to Daphnia magna and Nitocra spinipes. Immobility in Daphnia after exposure to 16 percent sediment (wet wt) in reconstituted standardized water (ISO, 1982) ranged from 0 to 88 percent after 24 h and from 3 to 95 percent after 48 h. For Nitocra the toxicity, determined as the 96-h LC50 (% wet wt) at 7 salinity, ranged from > > 32 percent (nontoxic) to 1.8 percent (most toxic). All exposures were made in duplicates and the effects obtained in the duplicates with the same sediment were correlated to each other. However, sediment toxicity to Daphnia and Nitocra was not. The test with Nitocra, which was made at several concentrations of sediment, was considered to give the most reliable picture of sediment toxicity in the Kattegat and Skagerrak. This ambient toxicity assessment identified three areas with toxic sediment, (1) the Göta älv estuary (outside the city of Göteborg) and its surroundings, (2) the Bay of Laholm in southern Kattegat, which is an area with periodic oxygen depletion and where repeated mussel kills have occurred during the last decade, and (3) an area in the open Skagerrak northwest of Skagen (the tip of the Jutland peninsula). Sediments, which had been stored at 4 °C, were tested again after 6 to 13 mos with the Nitocra test. Stored sediment toxicity was poorly correlated with fresh sediment toxicity. The average detoxification during storage was 5 times, but the range was 3 orders of magnitude, from 17 times more toxic to 73 times less toxic. The reasons for the observed areal and storage differences in sediment toxicity are so far not understood.     

An Integrated System for the Bioremediation of Wastewater Containing Xenobiotics and Toxcic Metals

An integrated system for the biotreatment of acidic wastewaters containing both toxic metals and organics is presented. It consists of two bioprocess stages (i) an anaerobic, SRB stage (containing alkaline‐tolerant s ulfate‐ r educing b acteria) that at pH 8 (chosen to acclimatize the bacteria in the biomedium) produces high concentrations of total sulfide ions (more than 400 mg/L) which are added to the wastewater to precipitate the heavy metals out at pH 2 as metal sulfides, and (ii) an aerobic, acidophilic stage containing heterotrophic bacteria (WJB3) that degrade organic xenobiotics. The anaerobic system was comprised of a 4‐L fluidized bed bioreactor with immobilized SRB, a mixing tank, and a precipitation tank. The effluent from the bioreactor with a high concentration of sulfide ions was fed into a mixing tank where model wastewaters containing toxic metals and phenol at pH 2 were also fed at increasing loading rates until free metal ions could be detected in the precipitation tank outlet. Then the effluent from the precipitation tank outlet was fed into a 2.5‐L aerobic bioreactor in which phenol was degraded. In this research, 100 % removal efficiencies were obtained with wastewaters containing more than 400 mg/L metal ions and 900 mg/L phenol at a 6‐h HRT of the mixing tank.     

Microbial validation of vent filters

The Upjohn Company uses filtration to remove microorganisms and particulates from air and other gases which may come in contact with sterile products. To validate the microbial retentivity of these filters, they were challenged with an aerosol of Bacillus subtilis var niger spores. An aerosol challenge was used because it more closely simulated the use for which these filters were designed. The test apparatus was constructed of autoclavable components using a jet-type nebulizer and heated air mixing tube. Characterization of the aerosol particle size distribution with a particle size analyzer demonstrated that 80% of the particles had a diameter of x 3.0 times;m and that the particles had a mean mass diameter of 1.9 times;m with a geometric standard deviation of 1.8 times;m. Studies conducted with aerosols of Bacillus subtilis var niger spores demonstrated that the test apparatus could recover ca. 50% of the spores that were aerosolized. Hydrophobic filters from various manufactures were challenged with an aerosol of at least 10(8) spores of Bacillus subtilis. All filters tested could retain at least 10(9) spores when physical integrity of the filter was verfield.     

2D exchange 31P NMR spectroscopy of bacteriophage M13 and tobacco mosaic virus.

Two-dimensional (2D) exchange 31P nuclear magnetic resonance spectroscopy is used to study the slow overall motion of the rod-shaped viruses M13 and tobacco mosaic virus in concentrated gels. Even for short mixing times, observed diagonal spectra differ remarkably from projection spectra and one-dimensional spectra. Our model readily explains this to be a consequence of the T2e anisotropy caused by slow overall rotation of the viruses about their length axis. 2D exchange spectra recorded for 30% (w/w) tobacco mosaic virus with mixing times < 1 s do not show any off-diagonal broadening, indicating that its overall motion occurs in the sub-Hz frequency range. In contrast, the exchange spectra obtained for 30% M13 show significant off-diagonal intensity for mixing times of 0.01 s and higher. A log-gaussian distribution around 25 Hz of overall diffusion coefficients mainly spread between 1 and 10(3) Hz faithfully reproduces the 2D exchange spectra of 30% M13 recorded at various mixing times in a consistent way. A small but notable change in diagonal spectra at increasing mixing time is not well accounted for by our model and is probably caused by 31P spin diffusion.     

一种改良的植物DNA提取方法

植物组织中含有大量多糖、多酚、酯类等次生代谢产物, 要从中提取高质量的DNA比较困难。针对这一情况, 该文提出一种改良CTAB植物DNA提取方法(mCTAB), 并以10种常见植物为实验材料, 与4种常用的植物DNA提取试剂盒作对比。结果表明, mCTAB法提取的DNA产率高且质量好, PCR扩增成功率也较高, 而提取成本显著低于DNA提取试剂盒, 可有效用于植物DNA条形码等研究的植物DNA提取。     

Measurement of axial diffusivities in a model of the bronchial airways.

Values for the effective axial diffusivity D for laminar flow of a gas species in the bronchial airways have been obtained as a function of the mean axial gas velocity u by experiment measurements of benzene vapor dispersion in a five generation glass tube model of the bronchial tree. For both inspiration and expiration D is seen to be approximately a linear function of u over the range of Reynolds' numbers 30-2,000 corresponding to peak flows in bronchial generations 0-13 under resting breathing conditions. The diffusivity for expiration is seen to be approximately one-third that for inspiration due presumably to increased radial mixing at bifurcations during expiration. The effective diffusivities relative to the molecular diffusivity can be expressed by the formulas D/Dmol = 1 + 1.08 NPe for inspiration and D/Dmol = 1 + .37 N-Pe for expiration. These velocity dependent diffusivities help to explain the short transit times of gas boluses from mouth to alveoli and will aid in the analysis of airway gas mixing by mathematical transport equations.     

Mixing analysis of PCS slurries in a horizontal scraped surface bioreactor

Horizontal rotating reactors offer many advantages for enzymatic hydrolysis of viscous biomass slurries; however, they do not provide homogenous mixtures since motion is only in the angular direction. Multi-directional mixing is important for dispersing enzymes and carrying products away from reaction sites. The objective here was to experimentally quantify mixing times and axial dispersion coefficients in a horizontal rotating bioreactor. Mixing times were of the same order as reaction times, indicating that enzymatic hydrolysis could be as much controlled by diffusion and mixing effects as by the complex reaction mechanism. The dispersion coefficient for the highest solids slurry was 20× less than the lowest solids slurry, which is indicative of the difference in free water and the magnitude change of viscosity with relatively small addition of solids. The slow mixing times and low dispersion may be an acceptable tradeoff with significantly lower power requirements compared to a conventional vertical reactor.