Enzymatic ester synthesis with continuous measurement of water activity

Ester synthesis from aliphatic monoalcohols and organic acids was investigated by using a microbial lipase. The reaction medium only contained the substrates and the enzyme without addition of water or organic solvent. During the reaction, water was produced and the water activity (aw) increased. Batch reactors and continuous-flow reactors were used. In batch, the aw was 0.13 at the beginning of the reaction and increased to reach a plateau at 0.77, after which ester synthesis continued without modification of the aw. Different alcohols and acids were tried in solid-liquid reactors, and all cases synthesis occurred, leading to a significant increase in the water activity. For continuous-flow reactors, the use of silica beads retaining water inside the reactor where the enzymatic reaction took place resulted in some control of the enzymatic reaction by changing the aw.     

Computer-aided control of water activity for lipase-catalyzed esterification in solvent-free systems

A computer system for on-line monitoring and control of the water activity (a(w)) in solvent-free media has been developed. The performance of this system was investigated by carrying out the lipase-catalyzed esterification of n-capric acid with n-decyl alcohol. A humidity sensor measured the relative humidity in the reactor headspace, which was then transmitted electrically to a digital computer that was used as a feedback controller. The water activity control was achieved by sparging either humidified air or dried air through the reaction medium at a flow rate determined by the digital feedback controller. The use of humid air and dry air for a(w) control made it possible to induce a larger a(w) gradient and thereby higher water transfer rate. As a result, the water activity quickly reached the desired a(w) values. We tested whether water activity in the reaction medium can be monitored by measuring relative humidity in the headspace. When the water activity in the liquid phase was determined from measurements of water content in the medium and compared to that measured directly with the humidity sensor, the a(w) in the reaction medium did not differ significantly from that in the headspace. This indicates that there is a near-equilibrium between the liquid medium and the exit air stream. Water activity was also successfully maintained close to the set point despite the massive production of water during the esterification process. Thus, the control system developed in this study is particularly useful for systems where large amounts of water are produced and where conventional methods make it difficult to control water activity as a result of a low water transfer rate. The effects that computer control of the water activity had on the reaction rate and yield were also examined. The reaction yield was significantly improved with water activity control. The conversions obtained at 28 h without and those with water activity control were 70% and 96%, respectively. In addition, from the fact that the final yields increased with decreasing a(w), computer-aided water activity control was performed with a set-point change. By controlling a(w) at 0.55 during initial reaction phase, followed by a step change of a(w) from 0.55 to 0 after 11 h of reaction, it was possible to enhance the final conversion to 100%.     

Effect of Temperature on Survival of Airborne Mycoplasma pneumoniae

Aerosols of Mycoplasma pneumoniae were prepared at each of eight relative humidities between 0 and 85% and at five separate temperatures between 10 and 43 C. Survival of these organisms was found to be a function of both relative humidity and temperature. However, the temperature response was mediated by humidity in that the effects of temperature could be observed only if some water vapor was present. At all temperatures, survival of M. pneumoniae in aerosols was found to be best at the extremes of relative humidity. The effects of temperature were such that irrespective of relative humidity an increase in temperature resulted in a decreased airborne survival time.     

Development of a Polyaniline-coated Monolith Reactor for the Synthesis of Cephalexin Using Penicillin G Acylase Aggregates

A monolith reactor for the synthesis of cephalexin was developed using capillary columns. The micro channel in the monolith reactor was coated with polyaniline (PANI), and penicillin G acylase was aggregated with PANI using 0.5% of glutaraldehyde as a cross-linker. The developed monolith reactor exhibited many advantages over other enzyme reactors such as batch and continuous reactors. It showed fast enzyme reaction rates owing to the decrease in external mass transfer and internal diffusion limitations. The reactor can easily be scaled up by bundling together multiple monolith reactors, enabling a corresponding increase in feed rate. Furthermore, the monolith reactor showed good operational stability, with 95% of its original activity maintained after 48 h of continuous operation. The PANI coating on the surface of the capillary column increased the enzyme immobilization capacity and conversion was increased from 15.4% to 70.6% after PANI coating. The conversion ratio increased to approximately 70.6% with an increase in residence time and reactor length.     

One-step acclimatization of plantlets using a mist reactor

A mist reactor was used to grow and acclimatize carnation plants in vitro without using ex vitro acclimatization techniques. The acclimatization protocol in the reactor consisted of altering the mist-on period during the course of the culture period and a stepwise reduction in the relative humidity surrounding the plants from 98% to 70% relative humidity (RH) during the final week of in vitro growth. After transfer and further growth in a greenhouse for 5 weeks, survival was 91% for plants grown in reactors, 81% from vented boxes, and 50% from unvented boxes. Ex vitro survival directly correlated with increased in vitro rooting and decreased hyperhydration. In vitro rooting also correlated with high-quality plants, but did not significantly correlate with low hyperhydration, as normal plants often lacked roots. After 5 weeks in the greenhouse, the quantity of mid- and high-quality plants obtained from reactors and ventilated boxes was similar. Conditions in the mist reactor can be manipulated to produce plants that are readily acclimatized and are equal or better in quality and yield than plants produced using conventional methods.     

Thermal balance in composting operations

Thermal balance analysis of composting of sewage sludge cakes was conducted for a laboratory-scale batch operated autothermal packed bed reactor and a continuously operated commercial open-bin reactor with a scoop mixer, and necessary equations were developed. Substitution of experimentally determined reaction rates into them confirmed the validity of these equations. The relative importance of each term relating to heat removal from the reactors was quantitatively evaluated. In the commercial reactor, heat removal was mainly due to water vaporization from composting material, while in the laboratory reactor heat loss from the reactor wall was the largest. In the laboratory reactor, the amount of evaporation was largest when the air flow rate was controlled to maintain temperature in the reactor to maximize the reaction rate.     

Regioselective enzymatic diol esterification in batch and fixed-Bed adsorptive reactors: experiments and modeling

The dynamic behavior of batch and fixed-bed adsorptive reactors is studied for the enzyme-catalyzed regioselective esterification of propionic acid and 2-ethyl-1,3-hexanediol in hexane. The reaction is equilibrium-limited with an apparent equilibrium constant of 0.6 +/- 0.1 at 22 degrees C. Moreover, accumulation of water produced in the reaction onto the biocatalyst causes a decrease in the catalytic activity. As a result, improvements in both reaction rate and final conversion can be achieved by operating in an adsorptive-reactor mode. Control of water in the reactor is achieved with a catalytically inert ion-exchange resin in Na-form. The resin prevents an excessive accumulation of water on the biocatalyst and reduces equilibrium limitations. The thermodynamic activity of water is identified as a key parameter for the design of such reactors. A mathematical model capable of predicting the water activity as a function of the varying concentrations of reactants and products is thus developed and found to successfully predict the experimental behavior observed in laboratory reactors. Substantial improvements in performance predicted by the model are seen experimentally in batch reactions and during the transient operation of continuous-flow fixed-bed reactors combining adsorptive and catalytic functions.     

Thermal and hygric physiology of Australian burrowing mygalomorph spiders (Aganippe spp.)

This study investigated the standard metabolic rate (SMR) and evaporative water loss (EWL) responses of three Australian trapdoor-constructing mygalomorph spider species, two undescribed arid-zone species (Aganippe ‘Tropicana A’ and A. ‘Tropicana B’) and a mesic-dwelling species (A. rhaphiduca) to acute environmental regimes of temperature and relative humidity. There were significant effects of species, temperature, and relative humidity on SMR. SMR was lower for A. raphiduca than both A. ‘Tropicana’ spp. with no difference between the two A. ‘Tropicana’ spp. Metabolic rate increased at higher temperature and relative humidity for all three species. There were significant effects of species, temperature, and relative humidity on EWL. The mesic Aganippe species had a significantly higher EWL than either arid Tropicana species. EWL was significantly higher at lower relative humidity. Our results suggest an environmental effect on EWL but not SMR, and that mygalomorphs are so vulnerable to desiccation that the burrow provides a crucial refuge to ameliorate the effects of low environmental humidity. We conclude that mygalomorphs are highly susceptible to disturbance, and are of high conservation value as many are short-range endemics.     

Optimization of a reactor assembly for the production of 6-APA from penicillin-V

The design and operation of an industrial penicillin-V deacylation reactor is simulated, using a kinetic expression and mass transport parameters for the immobilized enzyme particles which were determined experimentally in a previous study. It is desirable to use a series of equalsized plug flow reactors with pH control at the entrance to each reactor, and with a possibility of recycling reactant in each reactor. These measures are necessary to avoid a steep pH profile through the reactor; the deacylation reaction is accompanied by an increase of acidity of the reaction medium, and H(+) is a strong inhibitor and may deactivate the enzyme. The optimization study which is carried out at a fixed penicillin conversion of x = 0.99 shows that it is uneconomical to use penicillin feed concentrations above 150mM-175mM, and that the buffer concentration in the reaction medium should not be less than 50mM-75mM. Increasing the number of reactors from 4 to 8 or 10 leads to higher productivity of 6-APA, and a moderate recycle in the first couple of reactors diminishes the sharp decrease in pH which will be found in a straight plug flow reactor operation of the equipment. Higher pumping costs and lower productivity are unavoidable drawbacks of an operation mode where the separation costs for the product mixture are desired to be low.     

Startup of anaerobic downflow stationary fixed film (DSFF) reactors

One of the serious problems limiting the application of full-scale anaerobic fixed film processes is reactor startup. To better understand startup, studies with downflow stationary fixed film (DSFF) reactors were conducted to characterize the effects of influent concentration, support material, and surface-to-volume ratio on biofilm development and overall reactor performance. Materials with roughened surfaces gave the best startup performance and as expected increased surface area in the reactors led to more rapid increases in loading rates and higher ultimate loadings. Soluble influent COD concentrations between 5 x 10(3) and 2 x 10(4) mg/L influenced the rate of biofilm development. Lower COD concentrations resulted in faster development of the biofilm, even though ultimate loadings were not necessarily achieved as rapidly as in reactors fed higher strength wastes. No decrease in specific activity of the biofilms in each reactor was observed as the thickness of the biofilms increased to their maximum value at the ultimate loadings. The operation of reactors fed lower strength wastes was more stable than reactors receiving higher strength feeds at comparable loadings. Biofilm yield and activity, COD removals, suspended growth and activity, and other system parameters are discussed.     

Sows’ responses to increased heat load – A review

There is a comprehensive body of literature on how increased air temperature affects the physiology, production and behaviour of sows, while very few studies consider the thermal effects of air humidity and air velocity.This review summarises studies that have investigated effects of air temperature by reviewing published literature in which sows were exposed to at least two different levels of air temperature ranging from 15 °C to 39 °C. Increased rectal temperature was investigated in the majority of the studies (26) and on average, the rectal temperature increased by 0.099 °C per °C increased air temperature above 25 °C. The increase was smaller at lower air temperatures, and it was suggested that rectal temperature is practically unaffected by air temperatures in the range of 15 °C–21 °C. This review elucidates how air temperature also affects performance indicators such as respiration rate, vaginal temperature, skin temperature, feed intake, milk yield, body weight loss during lactation, mortality, litter daily weight gain during lactation and sow behaviour.One study reported how respiration rate, rectal temperature, vaginal temperature and skin temperature were affected by both air temperature and air humidity, and the results suggest that the relative significance of air temperature and humidity may be similar for sows and finishing pigs (e.g. an increase of 40% relative humidity at an air temperature of 30 °C has a similar effect as a 1.9 °C increase in temperature).Studies on mitigation methods against the effects of high temperature and humidity such as snout cooling, drip cooling and floor cooling were reviewed to extract knowledge related to the effects of air velocity, temperatures of surrounding surfaces and the opportunity for sows to moisten their skin.     

Optimization of Operating Temperature for Continuous Immobilized Glucose Isomerase Reactor with Pseudo Linear Kinetics

In this work, the optimal operating temperature for the enzymatic isomerization of glucose to fructose using a continuous immobilized glucose isomerase packed bed reactor is studied. This optimization problem describing the performance of such reactor is based on reversible pseudo linear kinetics and is expressed in terms of a recycle ratio. The thermal deactivation of the enzyme as well as the substrate protection during the reactor operation is considered. The formulation of the problem is expressed in terms of maximization of the productivity of fructose. This constrained nonlinear optimization problem is solved using the disjoint policy of the calculus of variations. Accordingly, this method of solution transforms the nonlinear optimization problem into a system of two coupled nonlinear ordinary differential equations (ODEs) of the initial value type, one equation for the operating temperature profile and the other one for the enzyme activity. The ODE for the operating temperature profile is dependent on the recycle ratio, operating time period, and the reactor residence time as well as the kinetics of the reaction and enzyme deactivation. The optimal initial operating temperature is selected by solving the ODEs system by maximizing the fructose productivity. This results into an unconstrained one‐dimensional optimization problem with simple bounds on the operating temperature. Depending on the limits of the recycle ratio, which represents either a plug flow or a mixed flow reactor, it is found that the optimal temperature of operation is characterized by an increasing temperature profile. For higher residence time and low operating periods the residual enzyme activity in the mixed flow reactor is higher than that for the plug flow reactor, which in turn allows the mixed flow reactor to operate at lower temperature than that of the plug flow reactor. At long operating times and short residence time, the operating temperature profiles are almost the same for both reactors. This could be attributed to the effect of substrate protection on the enzyme stability, which is almost the same for both reactors. Improvement in the fructose productivity for both types of reactors is achieved when compared to the constant optimum temperature of operation. The improvement in the fructose productivity for the plug flow reactor is significant in comparison with the mixed flow reactor.     

填充床反应器中酶法连续合成甘油二酯的研究

近年来,1,3-甘油二酯(DAG)由于其广泛用途及健康作用日益受到人们的重视。报道了一种无溶剂条件下填充床反应器中连续酶促合成1,3-DAG的方法。研究了填充柱的长径比、进料体积流速、温度、底物摩尔比对酯化率和1,3-DAG产量的影响。结果表明固定化酶填充柱长径比7.8,亚油酸、甘油摩尔比1∶2 ,进料速度1.2mL/min ,65℃条件下酯化反应可实现脂肪酸酯化率、1,3-DAG纯度及生产效率的统一。填充床反应器中固定化酶连续催化酯化反应的一个主要问题即体系水分清除困难。实验研究了采用过量甘油吸附脱水的可行性,亚油酸、甘油摩尔比为1∶2时,可明显改善固定化酶的稳定性,增加LipozymeRMIM的使用寿命。连续运行10d ,残余酶活仍保持在80 %以上,而对照组则仅为52%。     

A comparative study of immobilized amyloglucosidase in a packed bed reactor and a continuous feed stirred tank reactor

The catalytic activity of amyloglucosidase covalently attached to DEAE-cellulose was studied in a packed bed reactor and a continuous feed stirred tank reactor (CSTR) for the reaction maltose → glucose. At low flow rates mass-transfer limitations in the bed reactor lead to lower conversions for this reactor compared to the CSTR. Simple theoretical expressions for these reactors were compared with the experimental results. There are significant differences between the kinetic parameters and pH profile of the immobilized and free enzyme. The immobilized enzyme also showed greater stability at 50°C than did free amyloglucosidase. The temperature dependence of the reaction rate was the same for immobilized and free enzyme.     

Influence of climatic conditions on the mutations in pollen mother cells of Tradescantia clone 4430 and implications for the Trad-MCN bioassay protocol

The present was study aimed at investigating the influence of relative humidity and temperature on spontaneous and pollution-induced mutation rates during exposure and recovery periods in the Trad-MCN test. Cuttings of Tradescantia clone 4430 were exposed to a negative control, to 4 mM maleic hydrazide (MH), and to a polluted water sample under varying conditions of air temperature and humidity in climatic chamber experiments. The relative humidity did not affect the spontaneous mutation rate in the clone investigated, but was negatively correlated with the frequency of pollution-induced mutations. Low temperature caused an increase in the number of micronuclei in the negative control, but no comparable response in polluted samples. At an extremely high temperature, signs of strong physiological damage and/or of a meiotic delay of pollen maturation were detected. When the temperature increased gradually and the extreme value was maintained only for short time, such detrimental effects were not observed. Subsequent treatment with high and low temperatures, by contrast, resulted in the highest MCN rates of all experiments. Our studies point to the possibility of producing irregular results of the Trad-MCN test if the influence of climatic factors has not sufficiently been considered.     

Activity and storage stability of immobilized glucose oxidase onto magnesium silicate

Glucose oxidase (GOD) was covalently immobilized onto florisil (magnesium silicate) carrier via glutaraldehyde. Immobilization conditions were optimized: the amount of initial GOD per grams of carrier as 5 mg, pH as 5.5, immobilization time as 120 min and temperature as 10 °C. Under the optimized reaction conditions activities of free and immobilized GOD were measured. Free and immobilized GOD samples were characterized with their kinetic parameters, and thermal and storage stabilities. K_M and V_max values were 68.2 mM and 435 U mg GOD⁻¹ for free and 259 mM and 217 U mg GOD⁻¹ for immobilized enzymes, respectively. Operational stability of the immobilized enzyme was also determined by using a stirred batch type column reactor. Immobilized GOD was retained 40% of its initial activity after 50 reuses. Storage stabilities of the immobilized GOD samples stored in the mediums with different relative humidity in the range of 0–100% were investigated during 2 months. The highest storage stability was determined for the samples stored in the medium of 60% relative humidity. Increased relative humidity from 0% to 60% caused increased storage stability of immobilized GODs, however, further increase in relative humidity from 80% to 100% caused a significant decrease in storage stability of samples.     

Continuous in situ water activity control for organic phase biocatalysis in a packed bed hollow fiber reactor

Packed bed hollow fiber membrane reactors were used to carry out organic phase biocatalysis at constant water activity. The performance of the device was tested by carrying out the esterification of dodecanol and decanoic acid in hexane. Lipase from Candida rugosa, immobilized on microporous polypropylene and packed in the shell space of the reactor, was used to catalyze the reaction. In situ water activity control was accomplished by pumping appropriate saturated salt solutions through the microporous hollow fiber polypropylene membranes. Water generated by reaction in the organic phase, pumped continuously through the shell of the reactor, was transferred into the bulk of the aqueous phase under the water activity gradient. The reactor performance was found to be strongly dependent on the controlling water activity. By carefully selecting this control activity it was found possible to obtain complete esterification. The water activity of the organic phase could be maintained very close to that of the saturated salt solution used. The reactor could be operated in the continuous mode for 100 h without any degradation in its performance. (c) 1996 John Wiley & Sons, Inc.     

影响蝴蝶兰类原球茎耐脱水性的主要因素

为探讨蝴蝶兰(Phalaenopsis spp.)类原球茎(protocorm-like body,PLB)耐脱水性的主要影响因素,对PLB的平均粒重、含水率、脱水相对湿度、时间、温度、光周期与耐脱水性的关系进行了研究.结果表明,PLB的平均粒重与脱水后失水率、含水率、相对电导率、成活率呈显著或极显著相关.在较高湿度下...     

Changes in the structure and function of microbial communities in drinking water treatment bioreactors upon addition of phosphorus

Phosphorus was added as a nutrient to bench-scale and pilot-scale biologically active carbon (BAC) reactors operated for perchlorate and nitrate removal from contaminated groundwater. The two bioreactors responded similarly to phosphorus addition in terms of microbial community function (i.e., reactor performance), while drastically different responses in microbial community structure were detected. Improvement in reactor performance with respect to perchlorate and nitrate removal started within a few days after phosphorus addition for both reactors. Microbial community structures were evaluated using molecular techniques targeting 16S rRNA genes. Clone library results showed that the relative abundance of perchlorate-reducing bacteria (PRB) Dechloromonas and Azospira in the bench-scale reactor increased from 15.2% and 0.6% to 54.2% and 11.7% after phosphorus addition, respectively. Real-time quantitative PCR (qPCR) experiments revealed that these increases started within a few days after phosphorus addition. In contrast, after phosphorus addition, the relative abundance of Dechloromonas in the pilot-scale reactor decreased from 7.1 to 0.6%, while Zoogloea increased from 17.9 to 52.0%. The results of this study demonstrated that similar operating conditions for bench-scale and pilot-scale reactors resulted in similar contaminant removal performances, despite dramatically different responses from microbial communities. These findings suggest that it is important to evaluate the microbial community compositions inside bioreactors used for drinking water treatment, as they determine the microbial composition in the effluent and impact downstream treatment requirements for drinking water production. This information could be particularly relevant to drinking water safety, if pathogens or disinfectant-resistant bacteria are detected in the bioreactors.     

城市河道绿带宽度对空气温湿度的影响

以北京市北五环清河旁侧8段不同宽度的绿带为研究对象, 分别对样地和对照样地的气温和相对湿度进行同步测定, 每2 h测定一次, 每个季节连续测定7天。结果显示: 绿带在春、夏、秋季具有增湿降温效应, 并且此效应随着绿带宽度的增加而不断增强, 夏季的增湿降温效应最强, 春、秋季节的增湿降温效应差别不大; 绿带在冬季具有保温增湿效应, 随着绿带宽度的增加, 保温效应不断增强, 增湿效应不断减弱。四季中, 绿带温湿度变化受环境温度的影响较大, 在一天中的14:00-16:00时段, 春、夏、秋季增湿降温效应最强; 在此时段冬季的保温效应最弱, 增湿效应最强。