Incorporating hydrodynamics into spatiotemporal metabolic models of bubble column gas fermentation

Gas fermentation has emerged as a technologically and economically attractive option for producing renewable fuels and chemicals from carbon monoxide (CO) rich waste streams. LanzaTech has developed a proprietary strain of the gas fermentating acetogen Clostridium autoethanogenum as a microbial platform for synthesizing ethanol, 2,3-butanediol, and other chemicals. Bubble column reactor technology is being developed for the large-scale production, motivating the investigation of multiphase reactor hydrodynamics. In this study, we combined hydrodynamics with a genome-scale reconstruction of C. autoethanogenum metabolism and multiphase convection–dispersion equations to compare the performance of bubble column reactors with and without liquid recycle. For both reactor configurations, hydrodynamics was predicted to diminish bubble column performance with respect to CO conversion, biomass production, and ethanol production when compared with bubble column models in which the gas phase was modeled as ideal plug flow plus axial dispersion. Liquid recycle was predicted to be advantageous by increasing CO conversion, biomass production, and ethanol and 2,3-butanediol production compared with the non-recycle reactor configuration. Parametric studies performed for the liquid recycle configuration with two-phase hydrodynamics showed that increased CO feed flow rates (more gas supply), smaller CO gas bubbles (more gas–liquid mass transfer), and shorter column heights (more gas per volume of liquid per time) favored ethanol production over acetate production. Our computational results demonstrate the power of combining cellular metabolic models and two-phase hydrodynamics for simulating and optimizing gas fermentation reactors.     

Methane recovery from water hyacinth through whole-cell immobilization technology

The concepts of feed pretreatment, phase separation, and whole-cell immobilization technology have been incorporated in this investigation for the development of rational and cost-effective two- and three-stage methane recovery systems from water hyacinth (WH)Analyses of laboratory data reveal that a three-stage system could be designed with an alkali pretreatment stage [3.6% Na(2)CO(3) + 2.5% Ca(OH)(2) W/W, 24 h HRT] followed by an open acid reactor (2.1 days HRT) and closed immobilized methane reactor (12 h HRT), providing steady-state COD conversion of 62-65%, TVA conversion of 91-95%, and gas productivity of 4.08-5.36 L/L reactor volume/day with 82% methane. A gas yield of 50 L/kg WH/day (dry wt basis) at 35-37 degrees C is possible with this system. Insulation bricks, with particle size distribution of 500-3000 mum, were used as support material in the reactors at organic loading rate of 20 kg COD/m(3) day. The reactors matured in 15-18 weeksSubstantial reduction in retention time for the conversion of volatile acids in immobilized methane reactors prompted further research on the combined immobilized reactor to make possible an additional reduction in the cost of a WH-based biogas system. Evaluation of laboratory data reveals that a two-stage system could be designed with an open alkali pretreatment stage and a combined immobilized reactor (12 h HRT), providing steady-state COD conversion of 53% and gas productivity of 3.1 L/L reactor volume/day with 86% methane. A gas yield of 44 L/kg WH/day (dry wt basis) at 35-37 degrees C could be obtained from this system. Insulation bricks, with 500-1000 mum particle size distribution, was used as support material at an organic loading rate of 15 kg COD/m(3) day. Notwithstanding the fact that the technology in this study has been developed with water hyacinth as substrate, the implicit principles could be extended to any other organic substrate.     

Characterization of CIM monoliths as enzyme reactors

The immobilization of the enzymes citrate lyase, malate dehydrogenase, isocitrate dehydrogenase and lactate dehydrogenase to CIM monolithic supports was performed. The long-term stability, reproducibility, and linear response range of the immobilized enzyme reactors were investigated along with the determination of the kinetic behavior of the enzymes immobilized on the CIM monoliths. The Michaelis-Menten constant K(m) and the turnover number k(3) of the immobilized enzymes were found to be flow-unaffected. Furthermore, the K(m) values of the soluble and immobilized enzyme were found to be comparable. Both facts indicate the absence of a diffusional limitation in immobilized CIM enzyme reactors.     

Interaction between CO2 concentration and flow rate on peripheral airway resistance

In the present study, we investigated the interaction between CO2 concentration and rate of delivered flow on peripheral airway resistance (Rp) in the intact canine lung. Dogs were anesthetized, intubated, paralyzed, and mechanically ventilated with room air to maintain end-tidal CO2 between 4.8 and 5.2%. Using a wedged bronchoscope technique, we measured Rp at functional residual capacity. The relationship between CO2 concentration and Rp was measured at flow rates of 100 and 400 ml/min with 5, 3, 2, 1, and 0% CO2 in air. Measurements were made at the end of a 3-min exposure to each gas. At low flow rates (100 ml/min) responses to hypocapnia were small, whereas at high flow rates (400 ml/min) responses were large. The PC50 (defined as the CO2 concentration required to produce a 50% increase in Rp above baseline Rp established on 5% CO2) at 400 ml/min (1.73%) was significantly larger than that at 100 ml/min (0.38%). We also directly measured the relationship between Rp and flow rate with 5% CO2 (normocapnia) or 1% CO2 (hypocapnia) delivered into the wedged segment. Increases in normocapnic flow caused small but significant decreases in Rp. In contrast, increases in hypocapnic flow from 100 to 400 ml/min caused a 108% increase in Rp. Thus the response to hypocapnia is augmented by increasing flow rate. This interaction can be explained by a simple model that considers the effect of local ventilation-perfusion ratio and gas mixing on the local CO2 concentration at the site of peripheral airway contraction.     

Effect of biomass concentration and inoculum source on the rate of anaerobic cellulose solubilization

This study determines cellulose solubilization kinetics from controlled batch digestions and shows the effect of inoculum biomass concentrations. Separate measurements and analyzes were performed for sessile biomass (biofilms) and planktonic biomass (free suspensions). Experiments were conducted using either leachate enriched on cellulose or rumen fluid as inoculum to assess if the effect of biomass concentration was consistent for microbial populations from different source environments. All batch digestions were fitted to a first-order kinetic model (R² ranging from 0.94 to 0.99). Regression analysis used to compare the first-order hydrolysis rate showed that the first-order hydrolysis rate was most strongly correlated with the concentration of sessile biomass rather than with the concentration of total or planktonic biomass. The correlation between solubilization rate and sessile biomass was statistically the same for the rumen and leachate inoculated reactors indicating that at low concentration ratios of inoculum to cellulose, the rate of cellulose solubilization is dependant primarily on sessile biomass concentration rather than the species profile of the cellulolytic community.     

Immobilized microbes and a high-rate, continuous waste processor for the production of high Btu gas and the reduction of pollutants

Microbes capable of degrading organic wastes (sewage) are densely packed (immobilized) within the pores of controlled-pore ceramics. When the ceramic displays the optimum pore range for the reproduction of these microbes, the minimum volume for a very efficient reactor is required. A two-stage, anaerobic, immobilized microbe reactor has been designed, and laboratoryscale units have been constructed. A few of these units have been operated continuously for two years. These reactors were designed for the efficent conversion of carbon to methane in biodegradable molecules and for the effective transfer of that gas. The reactors were operated at 20, 30, and 40 degrees C at residence times of 2-5.5 h. The total chemical oxygen demand (COD) of the sewage varied from 800-2600 mg/L. The resulting gas contained greater than 90% methane and less than 5% CO(2). Approximately 32-54% of the influent total carbon was recovered as methane. The reduction in COD varied from 63 to 89%.     

Layered open pore poly(L-lactic acid) nanomorphology

This paper reports on specific open and interconnected CO(2) foams of poly(L-lactic acid). The effect of varying gas concentration and hence physical changes induced by CO(2) has been investigated and thus used to generate specific structures. The developed morphologies have a skin core structure with larger pores in the core and open and interconnected smaller pores in the skin.     

Inversion of sucrose in a continuous process with β- -fructofuranosidase (invertase) immobilized on bead DEAHP-cellulose

Inversion of sucrose with β- -fructofuranosidase (EC 3.2.1.26) immobilized by the ionic bond on bead DEAHP-cellulose has been studied under flow conditions. Under these conditions, the inversion of sucrose is affected by the concentration and flow rate of the substrate and by the reaction temperature. The effect of substrate concentration on the reaction was investigated in the range 19.5–64.2 wt %; the effect of flow rate was examined in the range 0.25–5.57 g solution per min, and the temperature range used was 25–50°C. It was found that the activities of immobilized β- -fructofuranosidase in stirred and flow reactors were the same. The lower activities of β- -fructofuranosidase in the case of concentrated solutions, and of immobilized β- -fructofuranosidase compared with the native enzyme are attributed to more difficult diffusion through the beads of the ion exchanger, especially of the strongly viscous substrate. A long-term investigation of the enzyme activity over a period of three months demonstrated the stability of the β- -fructofuranosidase immobilized by the ionic bond on bead DEAHP-cellulose; the half-life of the enzyme was 215 days. It was also found that the immobilization of the enzyme on a carrier was more effective under flow conditions, i.e. through an ion exchanger in the column, than under the equilibrium conditions of a stirred reactor.     

Feeding and respiratory gas exchange in the American dog tick,Dermacentor variabilis

This study investigated the effect of blood feeding on respiratory gas exchange in the dog tick Dermacentor variabilis. Adult male and female ticks were fed on bovine hosts from 1 to 11days. Females fed slowly for the first 6days and then rapidly engorged on blood 2-3days prior to dropping from the host. Ticks were removed at daily intervals during feeding, weighed and CO(2) emission measured at 25 degrees C using flow-through respirometry. During feeding, females (N=39) showed a 100-fold gain in mass from 5.78+/-1.05mg to 541.15+/-18.60mg while standard metabolic rate (Vdot;co(2)) increased from 0.179+/-0.030&mgr;lh(-1) in unfed ticks to 87.32+/-5.72&mgr;lh(-1) in fully engorged ticks. CO(2) release prior to feeding was highly discontinuous with discrete spiracular bursts of CO(2) emission approximately every 30min. For CO(2) emission measured in detached partially or completely fed ticks, burst frequency became more and more rapid as feeding progressed and changed to continuous sustained CO(2) output during rapid engorgement. In contrast to females, male ticks (N=20) showed little change in mass and maintained discontinuous CO(2) throughout the 11day attachment period on the host. The switch from discontinuous to continuous CO(2) release and presumed increase in respiratory water loss in female ticks is correlated to an increase in metabolic expenditure associated with blood meal digestion rather than any factor relating directly to maintenance of water balance.     

施肥对板栗林土壤CO2通量的影响

2011年6月-2012年6月,在浙江省临安市典型板栗林样地布置施肥试验,研究板栗林土壤CO₂通量与环境因子的关系.试验设置不施肥(对照)、施无机肥、有机肥及有机无机混合肥(1/2无机肥 + 1/2有机肥)4个处理.利用静态箱法测定土壤CO₂排放速率,以及土壤温度、含水量和水溶性有机碳(WSOC)含量.结果表明: 板栗林中土壤CO₂排放呈现显著的季节性变化特征,最小值均出现在2月,最大值均出现在7、8月.施用无机肥、有机肥和有机无机混合肥的土壤年累积CO₂通量比对照分别增加29.5%、47.0% 和50.7%.施用无机肥的土壤WSOC含量(105.1 mg·kg^-1)显著高于对照(76.6 mg·kg^-1),但明显低于有机肥(133.0 mg·kg^-1)和混合肥处理(121.17 mg·kg^-1).无机肥、有机肥和混合肥处理的土壤呼吸Q₁₀值(1.75、1.49和1.57)均高于对照(1.47).土壤CO₂排放速率与土壤5 cm温度、WSOC含量之间呈极显著正相关,但与土壤含水量没有明显的相关性.施肥导致土壤WSOC含量增加可能是板栗林地土壤CO₂排放速率增加的原因之一.     

Inversion of sucrose in a continuous process with β-d-fructofuranosidase (invertase) immobilized on bead DEAHP-cellulose

Inversion of sucrose with β-d-fructofuranosidase (EC 3.2.1.26) immobilized by the ionic bond on bead DEAHP-cellulose has been studied under flow conditions. Under these conditions, the inversion of sucrose is affected by the concentration and flow rate of the substrate and by the reaction temperature. The effect of substrate concentration on the reaction was investigated in the range 19.5–64.2 wt %; the effect of flow rate was examined in the range 0.25–5.57 g solution per min, and the temperature range used was 25–50°C. It was found that the activities of immobilized β-d-fructofuranosidase in stirred and flow reactors were the same. The lower activities of β-d-fructofuranosidase in the case of concentrated solutions, and of immobilized β-d-fructofuranosidase compared with the native enzyme are attributed to more difficult diffusion through the beads of the ion exchanger, especially of the strongly viscous substrate. A long-term investigation of the enzyme activity over a period of three months demonstrated the stability of the β-d-fructofuranosidase immobilized by the ionic bond on bead DEAHP-cellulose; the half-life of the enzyme was 215 days. It was also found that the immobilization of the enzyme on a carrier was more effective under flow conditions, i.e. through an ion exchanger in the column, than under the equilibrium conditions of a stirred reactor.     

Insights into large-scale cell-culture reactors: II. Gas-phase mixing and CO₂ stripping

Most discussions about stirred tank bioreactors for cell cultures focus on liquid-phase motions and neglect the importance of the gas phase for mixing, power input and especially CO(2) stripping. Particularly in large production reactors, CO(2) removal from the culture is known to be a major problem. Here, we show that stripping is mainly affected by the change of the gas composition during the movement of the gas phase through the bioreactor from the sparger system towards the headspace. A mathematical model for CO(2)-stripping and O(2)-mass transfer is presented taking gas-residence times into account. The gas phase is not moving through the reactor in form of a plug flow as often assumed. The model is validated by measurement data. Further measurement results are presented that show how the gas is partly recirculated by the impellers, thus increasing the gas-residence time. The gas-residence times can be measured easily with stimulus-response techniques. The results offer further insights on the gas-residence time distributions in stirred tank reactors.     

Fluorescent (Au@SiO2)SiC Nanohybrids: Influence of Gold Nanoparticle Diameter and SiC Nanoparticle Surface Density

Gold@silica core–shell nanoparticles were prepared with various gold core diameters (ranging from 20 to 150 nm) and silica thicknesses (ranging from 10 to 30 nm). When the gold diameter is increased, the size dispersion became larger, leading to a broader plasmon band. Then, silicon carbide (SiC) nanoparticles were covalently immobilized onto silica to obtain hybrid (Au@SiO₂) SiC nanoparticles. The absorption properties of these hybrid nanoparticles showed that an excess of SiC nanoparticles in the dispersion can be identified by a strong absorption in the UV region. Compared to SiC reference samples, a blue shift of the fluorescence emission, from 582 to 523 nm, was observed, which was previously attributed to the strong surface modification of SiC when immobilized onto silica. Finally, the influence of several elaboration parameters (gold diameter, silica thickness, SiC concentration) on fluorescence enhancement was investigated. It showed that the highest enhancements were obtained with 10 nm silica thickness, low concentration of SiC nanoparticles, and surprisingly, with a 20-nm gold core diameter. This last result could be attributed to the broad plasmon band of big gold colloids. In this case, SiC emission strongly overlapped gold absorption, leading to possible quenching of SiC fluorescence by energy transfer.     

Transport phenomena, reactor design and scale-up

This review will cover the area of impeller-mixed stirred-tank reactors. In addition, it will consider bubble columns, in which air or gas is passed up a liquid filled column through distribution plates covering the full area of the column, and also airlift reactors, in which the air is confined in a channel by means of a loop or draft tube designed to impart a certain type of overall circulatory pattern to the entire tank.There is considerable interest in the kinetics inside the solid part of various kinds of immobilized solid pellet type of enzymes and catalysts. The use of these particles in fixed bed reactors is also covered.     

Liquid residence time distributions in immobilized cell bioreactors

Previous work has demonstrated that high ethanol productivities can be achieved using yeast or bacterial cells adsorbed onto the surface of ion exchange resin in vertical packed bed bioreactors. The present work quantitatively characterizes the overall degree of backmixing in such reactors at two scales of operation: 2.0 and 8.0 L. Stimulus-response experiments, using two solvents (2,3-butanediol and 2-ethoxyethanol) as tracers, were performed to measure the liquid phase residence time distribution (RTD) during continuous ethanol fermentations using the yeast Saccharomyces cerevisiae and the bacterium Zymomonas mobilis at the 2-L scale, and with S. cerevisiae at the 8-L scale. In order to separately determine the effects of liquid flow rate and gas evolution on the degree of mixing, stimulus-response experiments were also performed in the systems without microbial cells present. The evolution of CO(2) was found to dramatically increase the extent of mixing; however, the tanks-in-series model for non-ideal flow represented the systems adequately. The packed beds were equivalent to over 70 tanks-in-series during abiotic operation while during fermentations, with similar liquid flow rates, they ranged in equivalence from 35 to 15 tanks-in-series. This increased knowledge of the overall degree of mixing in packed bed, immobilized cell bioreactors will allow for more accurate kinetic modelling and efficient scale up of the process.     

Anaerobic biodegradation tests and gas emissions from subsurface flow constructed wetlands

Anaerobic tests with gravel from horizontal subsurface flow constructed wetlands (SSF) used for the treatment of urban wastewater were developed in order to evaluate the anaerobic biodegradability of their effluents. Two types of assays were conducted. The reactors used for the first type were glass vials of 45 mL, that were used for only one measurement, requiring starting experiments with a number of reactors equal to the measurements to be made. For the second type of experiments multiple measurements were done in the same reactors, by using flasks of 2.2L. The COD of the SSF effluents used for the tests ranged from 60 to 130 mg/L. The evolution of CO(2) in the headspace of the reactors was used as indicator of anaerobic biodegradation rates. CO(2) mass emission rates ranged from 0.005 to 0.015 micromol/mL day. CH(4) generation was not detected in the tests in relation with the refractory properties of the effluent organic matter of the studied SSF. In situ measurement of CO(2) and CH(4) emissions from the gravel of the SSF ranged from 0.106 to 0.464 and from 0.039 to 0.107 mmol/m(2)h, respectively. Several CO(2) fluxes measured in the field were quite consistent with the emissions observed in the laboratory. The developed tests can help to understand the performance of SSF and improve their operation.     

The Effect of Mustard Gas on Salivary Trace Metals (Zn,Mn, Cu,Mg, Mo,Sr, Cd,Ca, Pb,Rb)

We have determined and compared trace metals concentration in saliva taken from chemical warfare injures who were under the exposure of mustard gas and healthy subjects by means of inductively coupled plasma optical emission spectroscopy (ICP-OES) for the first time. The influence of preliminary operations on the accuracy of ICP-OES analysis, blood contamination, the number of restored teeth in the mouth, salivary flow rate, and daily variations in trace metals concentration in saliva were also considered. Unstimulated saliva was collected at 10:00–11:00 a.m. from 45 subjects in three equal groups. The first group was composed of 15 healthy subjects (group 1); the second group consisted of 15 subjects who, upon chemical warfare injuries, did not use Salbutamol spray, which they would have normally used on a regular basis (group 2); and the third group contained the same number of patients as the second group, but they had taken their regular medicine (Salbutamol spray; group 3). Our results showed that the concentration of Cu in saliva was significantly increased in the chemical warfare injures compared to healthy subjects, as follows: healthy subjects 15.3± 5.45(p.p.b.), patients (group 2) 45.77±13.65, and patients (Salbutamol spray; group 3) 29 ±8.51 (P <0.02). In contrast, zinc was significantly decreased in the patients, as follows: healthy subjects 37 ± 9.03(p.p.b.), patients (group 2) 12.2 ± 3.56, and patients (Salbutamol spray; group 3) 20.6 ±10.01 (P < 0.01). It is important to note that direct dilution of saliva samples with ultrapure nitric acid showed the optimum ICP-OES outputs.     

Growth and biochemical characterization of microalgal biomass produced in bubble column and airlift photobioreactors: studies in fed-batch culture

Relatively large (0.19 m column diameter, 2 m tall, 0.06 m³ working volume) outdoor bubble column and airlift bioreactors (a split-cylinder and a draft-tube airlift device) were compared for monoseptic fed-batch culture of the microalga Phaeodactylum tricornutum. The three photobioreactors produced similar biomass versus time profiles and final biomass concentration (4 kg m⁻³). The maximum specific growth rate observed within a daily illuminated period in the exponential growth phase, had a value of 0.08 h⁻¹ on the third day of culture. Because of night-time losses of biomass, the specific growth rate averaged over the 4-days of exponential phase was 0.021 h⁻¹ for the three reactors.

The biomass in the vertical column reactors did not experience photoinhibition under conditions (photosynthetically active daily averaged irradiance value of 1150±52 μE m⁻² s⁻¹) that are known to cause photoinhibition in conventional thin-tube horizontal loop reactors. Because of good gas-liquid mass transfer, the dissolved oxygen concentration in the reactors at peak photosynthesis remained <120% of air saturation; thus, oxygen inhibition of photosynthesis and photo-oxidation of the biomass did not occur. Carbohydrate accumulation (up to 13% w/w) by the biomass was favored during light-limited linear growth. A declining light intensity caused a more than five-fold increase in cellular carotenoids but the chlorophylls increased only by about 2.5-fold during the course of the culture. In the stationary phase, up to 2% of the biomass was chlorophylls and carotenoids constituted up to 0.5% of the biomass dry weight.     

Removal of H2S in down-flow GAC biofiltration using sulfide oxidizing bacteria from concentrated latex wastewater

A biofiltration system with sulfur oxidizing bacteria immobilized on granular activated carbon (GAC) as packing materials had a good potential when used to eliminate H(2)S. The sulfur oxidizing bacteria were stimulated from concentrated latex wastewater with sulfur supplement under aerobic condition. Afterward, it was immobilized on GAC to test the performance of cell-immobilized GAC biofilter. In this study, the effect of inlet H(2)S concentration, H(2)S gas flow rate, air gas flow rate and long-term operation on the H(2)S removal efficiency was investigated. In addition, the comparative performance of sulfide oxidizing bacterium immobilized on GAC (biofilter A) and GAC without cell immobilization (biofilter B) systems was studied. It was found that the efficiency of the H(2)S removal was more than 98% even at high concentrations (200-4000 ppm) and the maximum elimination capacity was about 125 g H(2)S/m(3)of GAC/h in the biofilter A. However, the H(2)S flow rate of 15-35 l/h into both biofilters had little influence on the efficiency of H(2)S removal. Moreover, an air flow rate of 5.86 l/h gave complete removal of H(2)S (100%) in biofilter A. During the long-term operation, the complete H(2)S removal was achieved after 3-days operation in biofilter A and remained stable up to 60-days.     

Experimental and theoretical evidence for convective nutrient transport in an immobilized cell support.

Even though immobilized-cell reactors possess several engineering advantages over free-cell reactors, their full potential has not been realized because mass transfer often limits the rate of nutrient supply and product removal from immobilized cell supports. We studied the interaction between mass transfer and reaction kinetics in the anaerobic conversion of glucose to CO2 and ethanol by yeast immobilized in a porous rotating disk on the agitator shaft of a conventional CSTR. A Sherwood number correlation was used to show that external mass-transfer resistances were negligible under typical operating conditions. The modulus of Weisz based on observable reaction parameters was used to gauge the importance of pore diffusion limitations. Under conditions for which significant pore diffusion effects and hence low effectiveness factors (eta = ca. 0.1) would be predicted, the observed reaction rates were much higher than expected (eta = ca. 1), suggesting that pore diffusion limitations were at least partially relieved by convective transport of glucose into the support. Two possible mechanisms of convective transport are discussed. We hypothesize that gas evolution was responsible for the convective enhancement of glucose supply.