New technique for kla measurement between gas and water in aerated solvent-in-water dispersions

Summary A new technique is presented to determine gas-to-water overall volumetric mass transfer coefficients (k _l a) in a stirred-tank reactor containing solvent-in-water dispersions. The compound to be transferred from the gas to the water was toluene; the water-immiscible organic solvent was FC40, a perfluorocarbon. The k _l a was determined in steady-state conditions in the absence of biological consumption. Toluene removal was achieved by passing a continuous flow of toluene-free water through the reactor. When solvent was present it was separated from the water at the reactor outlet by means of a small settler and recycled back to the vessel. The k _l a was found to increase with the FC40 volume fraction. An enhancement of 1.9 times on an aqueous-phase-volume basis was found at 15 % (v/v) FC40.     

不同土壤含水量下侧柏幼树叶片水分利用效率

对植物水分利用效率的研究,可以揭示植物的内在耗水机制,为区域森林生态系统经营与维护提供依据.本研究以侧柏幼树为研究对象,通过室内控制试验设置不同的土壤水分梯度,分别用气体交换法和稳定同位素法对其不同土壤含水量条件下的瞬时水分利用效率(WUE_gs)和短期水分利用效率(WUE_cp)进行研究.结果表明:受气孔导度(g_s)的影响,叶片净光合速率(P_n)和蒸腾速率(T_r)随土壤含水量的增加呈现相同的变化趋势,均在土壤含水量为70%～80%田间持水量(FC)时达到最大值;叶片WUE_gs则在土壤含水量最低(35%～45% FC)时达到最大值(7.26 mmol·m^-2·s^-1).叶片可溶性糖、枝条韧皮部渗出液的δ¹³C都在土壤含水量最低(35%～45% FC)条件下达到最大值,且叶片可溶性糖的δ¹³C明显高于枝条韧皮部渗出液的δ¹³C,未产生明显分馏;而叶片WUE_cp也在土壤含水量最低(35%～45% FC)时达到最大值(7.26 mmol·m^-2·s^-1).相同条件下,叶片WUE_gs和WUE_cp存在一定差异(平均相差0.52 mmol·m^-2·s^-1),WUE_gs时空变异性较大,而WUE_cp更具有代表性.侧柏幼树通过降低生理生态活动和提高水分利用效率来适应干旱的土壤条件.     

Development of a novel bioreactor system for treatment of gaseous benzene

A novel, continuous bioreactor system combining a bubble column (absorption section) and a two-phase bioreactor (degradation section) has been designed to treat a gas stream containing benzene. The bubble column contained hexadecane as an absorbent for benzene, and was systemically chosen considering physical, biological, environmental, operational, and economic factors. This solvent has infinite solubility for benzene and very low volatility. After absorbing benzene in the bubble column, the hexadecane served as the organic phase of the two-phase partitioning bioreactor, transferring benzene into the aqueous phase where it was degraded by Alcaligenes xylosoxidans Y234. The hexadecane was then continuously recirculated back to the absorber section for the removal of additional benzene. All mass transfer and biodegradation characteristics in this system were investigated prior to operation of the integrated unit, and these included: the mass transfer rate of benzene in the absorption column; the mass transfer rate of benzene from the organic phase into the aqueous phase in the two-phase bioreactor; the stripping rate of benzene out of the two-phase bioreactor, etc. All of these parameters were incorporated into model equations, which were used to investigate the effects of operating conditions on the performance of the system. Finally, two experiments were conducted to show the feasibility of this system. Based on an aqueous bioreactor volume of 1 L, when the inlet gas flow and gaseous benzene concentration were 120 L/h and 4.2 mg/L, respectively, the benzene removal efficiency was 75% at steady state. This process is believed to be very practical for the treatment of high concentrations of gaseous pollutants, and represents an alternative to the use of biofilters.     

The Role of the Liquid Phase in the Degradation of Toluene and m‐Cresol within a Biofilm Trickle‐Bed Reactor

The degradation of toluene and m‐cresol in a biofilm trickle‐bed reactor was experimentally and theoretically investigated. Degradation is the result of the cooperation between suspended and immobilized microorganisms in the trickling film and the biofilm. The role of the trickling film is that of a barrier for mass transfer to the biofilm or that of an additional reaction space. This is the result of physical availability of pollutants to the liquid phase as well as co‐substrate degradation of inherent biomass. An instationary reactor balance model is presented. In addition to this the change in wetting behavior of carrier surface due to biofilm formation is discussed. A partial wetting of biofilm surface by rivulets of the trickling film is proposed. The model was verified by experimental data. The different reactor operation modes denoted as biofilm regime versus trickling film regime for the chosen pollutant system were expressed in terms of dimensionless reactions and transfer numbers. It is shown that the volumetric reaction rates for toluene in a trickling film regime reaches values twice as high as that of a biofilm regime due to the presence of the second substrate m‐cresol. The limiting step in both cases is the mass transfer of oxygen to bacteria in the biofilm or trickling film.     

Cyclodextrin-enhanced degradation of toluene and p-toluic acid by Pseudomonas putida.

Degradation of an immiscible aromatic solvent, toluene, and a water-soluble aromatic compound, p-toluic acid, by a Pseudomonas putida strain in the presence of beta-cyclodextrin (beta-CD) was investigated. The ability of CDs to interact with hydrophobic organics and form inclusion compounds was exploited in this study to remove or alleviate the toxicities of substrates and consequently to enable or enhance degradation. Liquid toluene was found to be highly toxic to P. putida. However, this phase toxicity was removed when crystalline beta-CD-complexed toluene was provided as the substrate. The latter was fully degraded at a concentration of up to 10 g/liter. Degradation of toluene vapors was enhanced in the presence of beta-CD as a result of reduced molecular toxicity and facilitated absorption of the gaseous substrate. Similarly, beta-CD alleviated the inhibitory effect of p-toluic acid on P. putida. This protective effect of CD was remarkably more prominent when the microbial culture was shock loaded with an otherwise toxic dose of p-toluic acid (1.8 g/liter).     

Carboxypeptidase Y-Catalyzed Reaction in Systems Containing Organic Solvent

The effect of organic solvents on carboxypeptidase Y (a serine carboxypeptidase from yeast)-catalyzed hydrolysis of amino acid ester and peptide synthesis from N-acyl amino acid ester and amino acid amide was investigated.

The K_m value of ester hydrolysis increased with an increase in the solvent content. Dioxane was the most effective and dimethyl sulfoxide (DMSO) the least, whilst K_cat showed a tendency to increase slightly in N, N-dimethylformamide (DMF) and DMSO. For dioxane and acetonitrile (MeCN) a maximum was observed.

In peptide formation from Fua-Phe-OEt and Gly-NH₂, dioxane and MeCN supported high product yield at molar fractions smaller than ca. 0.05 but the yield decreased significantly at higher fractions, although a relatively constant selectivity (ratio of the peptide bond formed to the ester consumed) was maintained. DMSO gave rather low peptide yields and selectivity even at lower molar fractions. DMF showed an intermediate tendency.

An apparent saturation parameter of the amine component was evaluated and the dissociation constant of a complex between acyl-enzyme and amino acid amide (K_n), as well as the rate constant of aminolysis exerted by the amino acid amide bound correctly on the enzyme (K_n), was calculated by initial rate analysis of peptide formation. In contrast to K_m values, K_n decreased with increasing concentrations of organic cosolvent. while a suppressive effect was observed (except for DMSO) on the K_n parameter.

Effects of the solvent practically immiscible in water was also studied by use of the enzyme physically “immobilized” on glass beads.     

A bioactive foam reactor for the removal of volatile organic compounds: system performance and model development

A bioactive foam reactor (BFR), a novel bioreactor operated using surfactant foams and suspended microorganisms for the treatment of gaseous toluene, was investigated to characterize its performance with respect to the mass transfer and biodegradation rates. The BFR system consisted of two reactors in series; a foam column for toluene mass transfer using fine bubbles and a cell reservoir where suspended microorganisms actively biodegraded toluene. In this study, a series of short-term experiments demonstrated that the BFR could achieve stable removal performance and a high elimination capacity (EC) for toluene at 100.3 g/m³/h. A numerical model, combining mass balance equations for the mass transfer and subsequent biodegradation, resulted in reasonable agreement with the experimental findings. At an inlet toluene concentration of 100 ppm_v, the toluene concentration in the liquid phase remained extremely low, indicating that the microbial activity was not hindered in the BFR system. However, the experimental and model prediction results showed that the actual mass of toluene transferred into the liquid phase was not closely balanced with the amount of toluene biodegraded in the BFR used in this study. Consequently, methods, such as increasing the effective volume of the foam column or the mass transfer coefficient, need to be implemented to achieve higher toluene EC and better BFR performance.     

Enhancement of the gas-to-water ethene transfer coefficient by a dispersed water-immiscible solvent: effect of the cells

  For a mass-transfer-limited system, it was demonstrated that the volumetric ethene transfer coefficient (k _l a) from gas to water could be enhanced by dispersing adequate amounts of a water-immiscible organic liquid, namely the perfluorocarbon FC40, in the aqueous phase. When 26% (v/v) FC40 was dispersed in a culture of Mycobacterium parafortuitum an enhancement of k _l a, calculated on a total liquid volume basis, of 1.8 times was found. Steady-state experiments in the absence of microorganisms, however, showed a 1.2-fold enhancement of k _l a at 18.5% (v/v) FC40. At all FC40 volume fractions tested, enhancement factors with cells were higher than enhancements without cells; apparently the microorganisms or their excretion products affected the interfacial areas or characteristic phase dimensions. Received: 4 December 1995 / Received revision: 7 June 1996 / Accepted: 10 June 1996     

A step-forward in the characterization and potential applications of solid and liquid oxygen transfer vectors

Silicone oil 20 and 200 cSt, a perfluorocarbon (FC40TM), heptamethylnonane, Kraton, Elvax, and Desmopan were evaluated for their ability to enhance oxygen transfer in stirred tank and airlift reactors (STR and ALR, respectively). None of the vectors tested was either toxic or biodegradable and they exhibited a moderate affinity for oxygen (gas/vector partitioning coefficients K_{\textg/\textv} = C_\textg ·C_\textv^{- 1} K_{{{\text{g}}/{\text{v}}}} = C_{\text{g}} \cdot C_{\text{v}}^{- 1} ranging from 3 to 5.1). FC40 was highly volatile, while KratonTM and ElvaxTM exhibited a low thermal stability, which constitutes a serious handicap for their implementation in fermentations. Silicone oil 20 cSt and Desmopan supported the highest oxygen transfer rates under abiotic conditions in both STR and ALR designs, with enhancement factors of up to 90% and 250%, respectively, compared to control tests (deprived of vector). The fact that these vectors showed the highest K _g/v proved that, besides the classical selection criteria, the in situ hydrodynamic behavior (which affects K _L a) must be considered for vector selection. The use of silicone oil 20 cSt and Desmopan in glucose-supplemented Saccharomyces cerevisiae fermentations resulted in a two- and threefold increase in biomass productions, respectively. The better performance of Desmopan in terms of biomass growth enhancement, together with the absence of the operational problems inherent to the use of liquid vectors (such as intensive foaming, high cost, and difficult solvent recovery), make solid vectors a promising and cost-effective alternative in the future developments of two-phase partitioning bioreactors.     

Mitochondrial functions under hypoxic conditions: The steady states of cytochrome c reduction and of energy metabolism

1. The effect of low oxygen concentration on the oxidation-reduction states of cytochrome c and of pyridine nucleotide, on Ca²⁺ uptake, on the energy-linked reduction of pyridine nucleotide by succinate, and on the rate of oxygen consumption have been examined under various metabolic conditions, using pigeon heart mitochondria.

2. The oxygen concentration required to provide half-maximal reduction of cytochrome c (p50_c) ranges from 0.27 to 0.03 μM (0.2-0.02 Torr) depending upon the metabolic activity. There is a linear increase of the p50_c value with increasing respiratory rate.

3. The fraction of the normoxic respiration that is observed at p50_c is 70–90% under State 4 conditions, but is 30% under State 3 conditions.

4. The oxygen requirement for half-maximal reduction of pyridine nucleotide (p50_PN) varies less than p50_c, being 0.08 μM in State 3 and 0.06 μM in the uncoupled state.

5. The ability of the mitochondria to exhibit an energy-linked reduction of pyridine nucleotide by succinate disappears at an oxygen concentration of 0.09 μM (0.06 Torr). Below this oxygen concentration, endogenous Ca²⁺ begins to be released from the mitochondria. Thus, the critical oxygen concentration for bioenergetic function of mitochondria corresponds approximately to 50% reduction of pyridine nucleotide (p50_PN).     

Liquid-to-Gas Mass Transfer in Anaerobic Processes: Inevitable Transfer Limitations of Methane and Hydrogen in the Biomethanation Process

Liquid-to-gas mass transfer in anaerobic processes was investigated theoretically and experimentally. By using the classical definition of k(L)a, the global volumetric mass transfer coefficient, theoretical development of mass balances in such processes demonstrates that the mass transfer of highly soluble gases is not limited in the usual conditions occurring in anaerobic fermentors (low-intensity mixing). Conversely, the limitation is important for poorly soluble gases, such as methane and hydrogen. The latter could be overconcentrated to as much as 80 times the value at thermodynamic equilibrium. Such overconcentrations bring into question the biological interpretations that have been deduced solely from gaseous measurements. Experimental results obtained in three different methanogenic reactors for a wide range of conditions of mixing and gas production confirmed the general existence of low mass transfer coefficients and consequently of large overconcentrations of dissolved methane and hydrogen (up to 12 and 70 times the equilibrium values, respectively). Hydrogen mass transfer coefficients were obtained from the direct measurements of dissolved and gaseous concentrations, while carbon dioxide coefficients were calculated from gas phase composition and calculation of related dissolved concentration. Methane transfer coefficients were based on calculations from the carbon dioxide coefficients. From mass balances performed on a gas bubble during its simulated growth and ascent to the surface of the liquid, the methane and carbon dioxide contents in the gas bubble appeared to be controlled by the bubble growth process, while the bubble ascent was largely responsible for a slight enrichment in hydrogen.     

Viscosity of locust bean, guar and xanthan gum solutions in the Newtonian domain: a critical examination of the log (ηsp)o-log c[η]o master curves

The viscosity in the low shear rate Newtonian domain of three biopolymers, locust bean gum, guar gum and xanthan gum was studied as a function of temperature and of polymer concentration in various aqueous solvents. The intrinsic viscosities [η]_o of both galactomannans are not modified in the presence of 10 or 40% sucrose. In this case, a master curve relating the Newtonian specific viscosity (η_sp)_o, to the reduced concentration c[η]_o is obtained and allows (in good agreement with theoretical conjectures), two critical concentrations C* and C** to be defined, from which the value of the expansion coefficient may be estimated. For xanthan, as expected for a polyelectrolyte, [η]_o depends strongly on salt concentration and on added sucrose and the results did not obey the above-mentioned master curve. However, it is shown that (η_sp)_o depends only on xanthan concentration whenC > C**, and then it is assumed that chain dimensions have attained their unperturbed values whatever the solvent. Considering that both types of chains, random coils (galactomannans) and semi-rigid (xanthan) should give the same (η_sp)_o-C[η]_o master curve for C > C** when [η]_o is replaced by its unperturbed counterpart [η]_θ, a method for estimating [η]_θ for the xanthan sample is proposed. In conclusion, the numerous exceptions to the widely accepted (η_sp)_o vs C[η]_o “universal” behaviour are mainly ascribed to significant differences in expansion coefficient values which depend on both the polymer and the solvent.     

Design of a control system for biotransformation of toxic substrates: toluene hydroxylation by Pseudomonas putida UV4

Using the hydroxylation of toluene to toluene cis-glycol by Pseudomonas putida UV4 as an example, the design of a feed-back control system for the addition of a toxic, poorly water-soluble substrate to a fed-batch biotransformation is described. In kinetic studies the reaction followed Michaelis-Menten behavior until toxic toluene concentrations were reached (2.4 mM), above which irreversible denaturation of the biocatalyst was observed. An algorithm, based on a system mass balance, was used to maintain the aqueous phase toluene concentration in the desired range for zero order kinetics. The mass balance required accurate and rapid analysis of the product and reactant in both the liquid and the vapor phase. Various analytical methods were considered and the effect of the sampling and analysis time on the response of the control system was examined.     

Toluene removal from waste air using a flat composite membrane bioreactor

In this report, gaseous toluene biodegradation results in a flat composite membrane reactor inoculated with Pseudomonas putida TVA8 are presented. Preliminary abiotic experiments showed that transport of toluene through the membrane was linearly and negatively correlated with the gas residence time (tau). During a 339-day biofiltration experiment, the influence of gas residence time (2-24 sec) and mass loading rate (B(v); 10-483 g x m(-3) h(-1)) on the toluene elimination capacity was investigated. A maximum elimination capacity (EC(max)) of 397 g x m(-3) h(-1) was achieved at tau = 24 sec and B(v) = 473 g x m(-3) h(-1). Expressed per unit membrane area, the EC(m,max) was 0.793 g x m(-2) h(-1), which is five times higher than results obtained with other membrane bioreactor experiments in the same range of loading rates. At low gas residence times, reactor performance was limited by mass transfer. Toluene concentration profiles along the membrane were measured for several biotic and abiotic conditions. For inlet concentrations (C(in)) up to 1 g x m(-3), more than 90% was eliminated at 15 cm from the reactor inlet. For C(in) > 1.65 g x m(-3), longer membranes are necessary to obtain these high removal efficiencies.     

A novel method of simulating oxygen mass transfer in two-phase partitioning bioreactors

An empirical correlation, based on conventional forms, has been developed to represent the oxygen mass transfer coefficient as a function of operating conditions and organic fraction in two-phase, aqueous-organic dispersions. Such dispersions are characteristic of two-phase partitioning bioreactors, which have found increasing application for the biodegradation of toxic substrates. In this work, a critical distinction is made between the oxygen mass transfer coefficient, k(L)a, and the oxygen mass transfer rate. With an increasing organic fraction, the mass transfer coefficient decreases, whereas the oxygen transfer rate is predicted to increase to an optimal value. Use of the correlation assumes that the two-phase dispersion behaves as a single homogeneous phase with physical properties equivalent to the weighted volume-averaged values of the phases. The addition of a second, immiscible liquid phase with a high solubility of oxygen to an aqueous medium increases the oxygen solubility of the system. It is the increase in oxygen solubility that provides the potential for oxygen mass transfer rate enhancement. For the case studied in which n-hexadecane is selected as the second liquid phase, additions of up to 33% organic volume lead to significant increases in oxygen mass transfer rate, with an optimal increase of 58.5% predicted using a 27% organic phase volume. For this system, the predicted oxygen mass transfer enhancements due to organic-phase addition are found to be insensitive to the other operating variables, suggesting that organic-phase addition is always a viable option for oxygen mass transfer rate enhancement.     

Measurement of steady-state values of respiration rate and oxidation levels of respiratory pigments at low oxygen tensions. A new technique

1. An apparatus was developed for the simultaneous measurement of steady-state values of respiration rate and oxidation level of respiratory pigments at low oxygen tensions. An open reaction system is utilized. The liquid sample is in contact with a gas mixture whose oxygen tension can be increased linearly with time at a rate so slow that the system is always practically at a steady state.

2. Assuming Michaelis-Menten kinetics in the respiration, theoretical curves for oxygen tension in the liquid and oxidation level of the terminal oxidase during a linear increase of the oxygen tension in the gas were calculated.

3. Measurements were performed on rat liver mitochondria. Steady-state curves for oxygen tension in the liquid and oxidation level of the terminal oxidase, cytochrome a₃, obtained with coupled mitochondria resembled the theoretical curves. For uncoupled mitochondria the cytochrome a₃ curve was signmoidal, deviating strongly from the theoretical curve.

4. The apparent K_m for oxygen uptake of coupled mitochondria in the presence of pyruvate and malate, in the absence of phosphate was found to be 0.5 μM. In the case of uncoupled mitochondria the oxygen tension in the liquid could not be measured with sufficient accuracy to allow comparison with Michaelis-Menten kinetics. The apparent K_m for oxygen uptake was less than 0.05 μM.     

Degradation of toluene/heptene mixtures in a trickling-bed bioreactor

Laws and regulations for the reduction of solvent emissions tend to force increasingly lower limits on gaseous emissions. Biological waste-gas treatment is an ecological and rather cheap method for solvent reduction, which is often used for waste-gas streams containing two or more components. The microbiological degradation of toluene and heptane as well as of several mixtures of these substances were investigated in a laboratory plant trickling-bed reactor. It was found that toluene was degraded at up to three times the rate for the heptane. Heptane elimination in the mixtures was low and the toluene elimination rate at low organic loads approached 100%. *** DIRECT SUPPORT *** AG903066 00010     

An MCD spectroscopic study of the molybdenum active site in sulfite oxidase: insight into the role of coordinated cysteine

Temperature-dependent magnetic circular dichroism (MCD) spectroscopy has been used for the first time to probe the electronic structure of the Mo active site in sulfite oxidase (SO). The enzyme was poised in the catalytically relevant [Mo(V):Fe(II)] state by anaerobic reduction of the enzyme with the natural substrate, sulfite, in the absence of the physiological oxidant cytochrome c. The [Mo(V):Fe(II)] state is of particular importance, as it is proposed to be a catalytic intermediate in the oxidative half reaction, where SO is reoxidized to the resting [Mo(VI):Fe(III)] state by two sequential one-electron transfers to cytochrome c. The MCD spectrum of the enzyme shows no charge transfer transitions below 17 000 cm⁻¹. This has been interpreted to result from (1) a severe reduction in ene-1,2-dithiolate sulfur in-plane and out-of-plane p orbital mixing, (2) a decrease in the dithiolate sulfur out-of-plane p-Mo d_xy orbital overlap, and (3) an orthogonal orientation between the vertical cysteine sulfur p (perpendicular to the Mo–S_cys σ-bond) and Mo d_xy orbitals. The spectroscopically determined cysteine sulfur p-Mo d_xy bonding scheme in the [Mo(V):Fe(II)] state is consistent with the crystallographically determined O–Mo–S_cys–C dihedral angle of 90° and precludes a covalent interaction between the vertical cysteine sulfur p orbital and Mo d_xy, effectively decoupling the cysteine from an effective through-bond electron transfer pathway. We have tentatively assigned a 22 250 cm⁻¹ positive C-term feature in the MCD as the cysteine S(σ)→Mo d_xy charge transfer that becomes allowed by a combination of configuration interaction and low-symmetry; however, the orbital overlap is anticipated to be quite small due to the near orthogonality of these orbitals. Therefore, we propose that the primary role of the coordinated cysteine is to decrease the effective nuclear charge on Mo by charge donation to the metal, statically poising the active site at more negative reduction potentials during electron transfer (ET) regeneration. Finally, the results of this study are consistent with the pyranopterin ene-1,2-dithiolate acting to couple the Mo site into efficient superexchange pathways for ET regeneration following oxygen atom transfer to the substrate.     

Use of an SF6-Based Diagnostic Tool for Assessing Air Distributions and Oxygen Transfer Rates during IAS Operation

A diagnostic test designed to assess air distribution and oxygen delivery rate to the aquifer during in situ air sparging (IAS) is described. The conservative tracer gas, sulfur hexafluoride (SF₆), is added upstream of the air injection manifold during steady IAS operation and groundwater samples are collected from the target treatment zone after some time period (usually 4 to 24 h). The appearance of SF₆ in groundwater is used to characterize the air distribution in the target treatment zone, while the SF₆ concentration increase with time is used to assess oxygen transfer rates to the target treatment zone. Conversion from SF₆ concentration to oxygen mass transfer rate involves correcting the SF₆ concentration increase over time for differences in the relevant chemical properties and injection air concentration. Data presented from a field demonstration site illustrate the utility of this test for identifying air distribution details not readily identified by deep vadose zone helium and groundwater pressure transducer response tests. Oxygen transfer rates at this site ranged from 0 to 20 mg-O₂/L-H₂O/d. Finally, a comparison of short-term SF₆ test data with longer-term dissolved oxygen data illustrated this test's utility for anticipating long-term dissolved oxygen distributions.