Dichloromethane removal from waste gases with a trickle-bed bioreactor

A 66 dm³ trickle-bed bioreactor was constructed to assess the possibilities of eliminating dichloromethane from industrial waste gases. The trickle-bed bioreactor was filled with a randomly-stacked polypropylene packing material over which a liquid phase was circulated. The pH of the circulating liquid was externally controlled at a value of 7 and the temperature was maintained at 25 °C. The packing material was very quickly covered by a dichloromethane-degrading biofilm which thrived on the dichloromethane supplied via the gas phase. The biological system was very stable and not sensitive to fluctuations in the dichloromethane supply. Removal of dichloromethane from synthetic waste gas was possible down to concentrations well below the maximal allowable concentration of 150mg/m³ required by West-German law for gaseous emissions. At higher dichloromethane concentrations specific dichloromethane degradation rates of 200 g h^–1 m^–3 were possible. At very low inlet concentrations, dichloromethane elimination was completely mass transfer limited.The gas-phase mixing could be described by a series of 10 to 7 identical ideally-mixed tanks for superficial gas velocities ranging from 150 to 450 m/h. Dichloromethane elimination with the tricklebed bioreactor was modelled using an overall mass-transfer coefficient that was dependent on the gas and liquid velocities. Masstransfer resistance within the biofilm was also accounted for. Using the model, elimination efficiencies were predicted which were very close to the experimentally observed values.     

Biofilm Growth and Bed Fluidization in a Fluidized Bed Reactor Packed with Support Materials of Low Density

Support materials of low‐density for fluidized bed reactors provide several operational advantages, including lower energy requirements and proper biofilm growth balance. The aim of this investigation was to study the extent of biofilm growth and bed fluidization in an experimental reactor, using polyester resin (ρ_pr = 1220 kg/m³) and vitrified expanded perlite (ρ_vep = 1710 kg/m³) as alternative support materials to conventional silica sand. A noteworthy amount of biofilm was observed to be attached to both support materials from the very beginning of the bioreactor operation. Nevertheless, there were significant variations in biofilm growth and activity over the course of the experimental trials. For both perlite and polyester beds, the highest biofilm mass and the highest total number of mesophilic bacteria were observed between the 7^th and the 10^th day, showing a steady state trend at the end of the experimental runs. The chemical oxygen demand (COD) removal levels were concomitant with biofilm mass and total mesophilic bacteria changes, although the polyester bed efficiency was slightly higher than that for the perlite bed. As expected, the polyester bed was fluidized at a lower re‐circulation flow compared to the perlite bed. Reactor back‐washing was not required for these support materials since biomass excess was adequately separated by means of a special internal device. The efficiencies of removal of organic matter achieved were acceptable (up to 78 %) despite the low volume of the support material (25 %) and the low hydraulic retention time (30 min).     

Heterotrophic cultivation of Paracoccus denitrificans in a horizontal rotating tubular bioreactor

In this work, the heterotrophic cultivation of bacterium Paracoccus denitrificans has been studied in a horizontal rotating tubular bioreactor (HRTB). After development of a microbial biofilm on the inner surface of the HRTB, conditions for one-step removal of acetate and ammonium ion were created. The effect of bioreactor process parameters [medium inflow rate (F) and bioreactor rotation speed (n)] on the bioprocess dynamics in the HRTB was studied. Nitrite and nitrogen oxides (NO and N₂O) were detected as intermediates of ammonium ion degradation. The biofilm thickness and the nitrite concentration were gradually reduced with increase of bioreactor rotation speed when the medium inflow rate was in the range of 0.5–1.5 l h⁻¹. Further increase of inflow rate (2.0–2.5 l h⁻¹) did not have a significant effect on the biofilm thickness and nitrite concentration along the HRTB. Complete acetate consumption was observed when the inflow rate was in the range of 0.5–1.5 l h⁻¹ at all bioreactor rotation speeds. Significant pH gradient (cca 1 pH unit) along the HRTB was only observed at the highest inflow rate (2.5 l h⁻¹). The results have clearly shown that acetate and ammonium ion removal by P. denitificans can be successfully conducted in a HRTB as a one-step process.     

Effects of flow rate and substrate concentration on the formation and H2 production of photosynthetic bacterial biofilms

Photosynthetic bacteria (PSB), Rhodopseudomonas palustris CQK 01, were immobilized on the surface of a thin glass slide in a lab-scale flat panel photobioreactor under different flow rates and substrate concentrations. The morphology, dry weight and thickness of the mature PSB biofilms were determined to reveal the relationship between biofilm formation and hydrogen production performance. The mature biofilm formed at a low flow rate and a high substrate concentration showed a looser structure, these structures of the mature biofilm then affected the H₂ production performance of the bioreactor during mature stage. The biofilm formed at a flow rate of 228 ml/h and a substrate concentration of 60 mmol/l exhibited the highest dry weight and optimally porous structure, which is beneficial not only for hydrogen removal from the biofilm but also glucose diffusion into the biofilm, thus significantly boosting the photo-hydrogen production performance.     

Gas-phase methyl ethyl ketone biodegradation in a tubular biofilm reactor: microbiological and bioprocess aspects

A novel type of bioreactor was designed to clean VOCs-containing air.The operation of this reactor consists in mixing the polluted gas and a mistof nutrient solution in the presence of microorganisms in order to maximizecontact and transfer between gas, liquid and microorganisms and to promotethe degradation kinetics and the relative removal efficiency of thepollutant. A bacterial consortium acclimatized to MEK and containing apreponderance of Alcaligenes denitrificans was established under non-axenicconditions. On the tubular reactor's glass walls, a continuous biofilm wasdeveloped. This biofilm was rapidly contaminated by two fungi able todegrade MEK: Geotrichum candidum and Fusarium oxysporum. Their abundance inthe reactor is probably linked to the acidic conditions inside the biofilmand to their broader tolerance for low pH values concomitant with MEKdegradation. In the reactor, a maximum volumetric degradation rate of 3.5 kgMEK/m³ _reactor·d was obtained for arelative removal efficiency of 35%, whereas the latter was maintainedat 70% for more modest applied loadings of 1.5 kgMEK/m³ _reactor ·d. In liquid batchcultures, a biomass originating from the biofilm was able to degrade 0.40g_MEK/g_DCW·h at the optimal pH of 7. Aregular cycle of detachment-recolonization was observed during the operationof the bioreactor. The maximal degradation activity was obtained with a thinbiofilm and was not increased as the biofilm grew in thickness. The overalldegradation rate of the process did not appear to be limited by thediffusion of oxygen inside the biofilm. Over short periods of time, the MEKtransfer from the gaseous phase to the biofilm was neither affected by thepresence of the mist nor by the wetting of the biofilm. A better control ofthe biofilm pH led to improved performance in terms of removal rate but notin terms of relative elimination efficiency.     

Biological decomposition of dichloromethane from a chemical process effluent

The application of specialized microorganisms to treat dichloromethane (DM) containing process effluents was studied. An aerobic fluidized bed reactor with a working volume of 801 filled with sand particles as carriers for the bacteria was used. Oxygen was introduced into the recycle stream by an injector device. DM was monitored semi-continuously. A processor controlled the feed volume according to the DM effluent concentration. Mineralization rates of 12 kg DM/m_bioreactor ³ · d were reached within about three weeks using synthetic wastewater containing 2000 mg/l DM as single carbon compound. DM from process water of a pharmaceutical plant was reduced from about 2000 mg/l in the feed to below 1 mg/l in the effluent at volumetric loading rates of 3 to 4 kg DM/m_bioreactor ³ · d. Degradation of wastewater components like acetone and isopropanol were favoured, thus making the process less attractive for waste streams containing high amounts of DOC other than of DM. DM concentrations of up to 1000 mg/l were tolerated by the immobilized microorganisms and did not influence their DM degradation capacity. The ability to mineralize DM was lost when no DM was fed to the reactor for 10 days.     

Effects of stratification in a fluidized bed bioreactor during treatment of metalworking wastewater

During wastewater treatment, biofilm-coated sand particles stratified in a fluidized bed bioreactor (FBB); particles coated by thicker biofilm segregated toward the top of the bed. Stratification was so well developed that at least two co-existing regions of significantly different mean biofilm thickness were visually distinct within the operating FBB. The observed stratification is attributed to differences in forces of drag, buoyancy, shear, and collisional impact, as well as differences of collision rate within the different regions. Particles with thick biofilm (thickness >100 μm) near the top of the bed consumed substrate at significantly lower rates per unit biomass than particles with thin biofilm (10-20 μm) near the bottom of the bed, thereby suggesting that substrate mass-transfer resistance through biofilm may limit biodegradation rates in the upper portion of the FBB. Large agglomerates of biomass floc and sand, which formed at the top of the fluidized bed, and sand particles with thick biofilm were susceptible to washout from the FBB, causing operational and treatment instability. Radial injection of supplemental liquid feed near the top of the bed increased shear and mixing, thereby preventing formation and washout of agglomerates and thickly coated sand particles. Supplemental liquid injection caused the mean specific biomass loading on the sand to increase and also increased the total biomass inventory in the FBB. Rates of biodegradation in the FBB appeared to be limited by penetration of substrates into the biofilm and absorption of oxygen from air into the wastewater. Copyright 1999 John Wiley & Sons, Inc.     

Reductive dechlorination of 2-chlorophenol in a hydrogenotrophic, gas-permeable, silicone membrane bioreactor

A gas-permeable silicone membrane bioreactor was used to cultivate the biofilm under hydrogenotrophic condition for reductive dechlorination of 2-chlorophenol (2-CP). The anaerobic sludge obtained from a swine wastewater treatment plant was immobilized by polyvinyl alcohol (PVA) so as to form a biofilm on the surface of the silicone tube. After acclimating for about 4 months, the bioreactor showed a high dechlorinating performance. Under the condition of continuous feeding with 2-CP at 25 mg/l and the hydraulic retention time of 15 h, the 2-CP removal efficiency reached 92.8% (2-CP decay rate: 0.67 g/m² d of surface area of silicone tube). H₂ was used as electron donor for dechlorinating 2-CP, and produced the dechlorinating intermediate, phenol. Both nitrate and sulfate played important roles in inhibiting 2-CP dechlorination through different biological mechanisms. Nitrate can be easily utilized as an electron acceptor by the biofilm, while sulfate cannot. Results of this study demonstrated that nitrate competed with 2-CP as the electron acceptor, while sulfate retarded the activity of hydrogen-dechlorinating bacteria and thus inhibited the 2-CP dechlorination.     

Characteristics of draft tube gas-liquid-solid fluidized-bed bioreactor with immobilized living cells for phenol degradation

Biological phenol degradation in a draft tube gas-liquid-solid fluidized bed (DTFB) bioreactor containing a mixed culture immobilized on spherical activated carbon particles was investigated. The characteristics of biofilms including the biofilm dry density and thickness, the volumetric oxygen mass transfer coefficient, and the phenol removal rates under different operating conditions in the DTFB were evaluated. A phenol degradation rate as high as 18 kg/m(3)-day with an effluent phenol concentration less than 1 g/m(3) was achieved, signifying the high treatment efficiency of using a DTFB.     

Dichloromethane utilized by an anaerobic mixed culture: acetogenesis and methanogenesis

Dichloromethane (8.9 mg/l) was eliminated from industrially polluted, anaerobic groundwater in a fixed-bed reactor (43 m³) which was packed with activated charcoal and operated continuously for over three years. The elimination of dichloromethane over this period was some ten-fold in excess of the sorptive capacity of the charcoal, and the elimination (3.7 mg/h·[kg of charcoal]: residence time, 49 h) was tentatively attributed to dehalogenative microorganisms immobilized on the charcoal. Anaerobic enrichment cultures, with dichloromethane as the sole added source of carbon and energy, were inoculated with material from the reactor. Reproducibly complete substrate disappearance in subcultures was observed when traces of groundwater (1%) or yeast extract (0.01%) were supplied. Fed-batch experiments under an atmosphere of CO₂ plus N₂ led to the conversion in 11 days of 11 mM dichloromethane to 3 mM acetate and 2 mM methane, with a growth yield of 0.4 g of protein/mol of dichloromethane; insignificant amounts (<1 M) of chloromethane accumulated. Methanogenesis could be inhibited by 50 mM 2-bromoethane sulfonate without any effect on the dehalogenation rate. The maximum dehalogenation rate was 0.13 mmol dichloromethane/h·l (2.6 mkat/kg of protein).Abbreviation DCM dichloromethane     

Cloning and characterization of dichloromethane dehalogenase from Methylobacterium rhodesianum for dichloromethane degradation

The gene encoding dichloromethane dehalogenase from Methylobacterium rhodesianum was cloned. Bioinformatic analysis showed that dichloromethane dehalogenase gene sequence from M. rhodesianum is almost identical to the one from Methylobacterium extorquens, with only one base difference. Dichloromethane dehalogenase was subsequently expressed in Escherichia coli BL21 (DE3) and purified. It was found that enzyme activity in recombinant cells was 3 times higher than that in the wild-type M. rhodesianum. Further investigation showed that recombinant dichloromethane dehalogenase was most active at 40°C at pH 7–8, and its K_M was 10.96 mM when treated with dichloromethane as substrate. The fitted curve of dichloromethane degradation gave a V_max of 0.43 mM/h of in 0.01 M phosphate buffer. Degradation efficiency of dichloromethane reached 86.11% within 20 h. In addition, it was found that degradation efficiency of dichloromethane was highly associated with glutathione concentration, supporting the reports that glutathione functions as coenzyme of dichloromethane dehalogenase for dichloromethane degradation.     

The influence of NaCl on the degradation rate of dichloromethane byHyphomicrobium sp.

The degradation of dichloromethane by the pure strainHyphomicrobium GJ21 and by an enrichment culture, isolated from a continuously operating biological trickling filter system, as well as the corresponding growth rates of these organisms were investigated in several batch experiments. By fitting the experimental data to generally accepted theoretical expressions for microbial growth, the maximum growth rates were determined. The effect of NaCl was investigated at salt concentrations varying from 0 to 1000 mM. Furthermore the dichloromethane degradation was investigated separately in experiments in which a high initial biomass concentration was applied. The results show that microbial growth is strongly inhibited by increased NaCl concentrations (50% reduction of _max at 200–250 mM NaCl), while a certain degree of adaptation has taken place within an operational system eliminating dichloromethane. A critical NaCl concentration for growth of 600 mM was found for the microbial culture isolated from an operational trickling filter, while a value of 375 mM was found for the pure cultureHyphomicrobium GJ21. The substrate degradation appears to be much less susceptible to inhibition by NaCl. Even at 800 mM NaCl relatively high substrate degradation rates are still observed, although this process is again dependent on the NaCl concentration. Here the substrate elimination is due to the maintenance requirements of the microorganisms. The inhibition of the dichloromethane elimination was also investigated in a laboratory scale trickling filter. The results of these experiments confirmed those obtained in the batch experiments. At NaCl concentrations exceeding 600 mM a considerable elimination of dichloromethane was still observed for during several months of operation. These observations indicate that the inhibition of microbial growth offers a significant control parameter against excessive biomass growth in biological trickling filters for waste gas treatment.     

Biological phenol degradation in a gas-liquid-solid fluidized bed reactor

Biological phenol degradation was performed experimentally in a gas-liquid-solid fluidized bed bioreactor using a mixed culture of living cells immobilized on activated carbon particles. A comprehensive model was developed for this system utilizing double-substrate limiting kinetics. The model was used to simulate the effects of changing inlet phenol concentration and biofilm thickness on the rate of biodegradation for two different types of support particles. The model shows that gas-liquid mass transfer is the limiting step in the rate of phenol biodegradation when the phenol loading is high.     

Heterogeneous biofilm activity in a segregated anaerobic fluidized bed bioreactor

Bed segregation in a fluidized bed bioreactor profoundly influenced biofilm thickness and microbial activities of the biofilm along the bed height. Bioparticles coated with a thin biofilm, observed at the bottom of the reactor, had a higher specific activity in propylene glycol and n-propanol degradation than in thick biofilms developed at the top of the reactor. Although no significant difference was observed in specific activity for propionate and acetate along the reactor flow axis, more total propionate and acetate conversion occurred in regions of thicker biofilm accumulation.     

A new bioreactor with a semi-fixed packing: investigation of degradation of phenol

A new bioreactor using a semi-fixed packing of frames with “sacks” made of a fabric of “Raschell” type stretched on them is proposed. The construction provides not only a large surface area of the biofilm carrier per unit volume of the apparatus, but also the possibility for an easy removal of the biomass after reaching a certain thickness of the biofilm increasing the gas velocity. Aerobic degradation of phenol in the new bioreactor, using microorganisms of the strain Pseudomonas putida, was studied. The experiments are carried out using water containing 0.7?g/l of phenol at a temperature of 27–30?°C. Different specific surface area of the packing (within 176 and 387?m²/m³) are studied. Degradation rates from 60 to 140?mg/(1?h) are attained. A retardation of the process at the end is observed, probably due to inhibition effect. This rate is 5–6 times higher than the rate observed when using free cells. At air velocity of 0.03–0.035?m/s (related to the total cross section of the bioreactor) the vibrations of the packing material lead to destruction and removal of the old biomass.     

Anaerobic fermentative production of lactic acid using cheese whey and corn steep liquor

Cheese whey was the most suitable substrate for production of lactic acid under anaerobic conditions by Entercoccus flavescens which, on supplementating with corn steep liquor (5% v/v) and 10 mM CaCO₃ at pH 5.5, 37°C, yielded 12.6 g lactic acid/l in 36 h. Production was scaled up to a 10 l bioreactor under controlled pH and continuous CO₂ supply and gave 28 g lactic acid/l in 30 h resulting in a net 8.7-fold increase in production as compared to unoptimized conditions.     

Toluene degradation kinetics for planktonic and biofilm-grown cells of Pseudomonas putida 54G

Toluene degradation kinetics by biofilm and planktonic cells of Pseudomonas putida 54G were compared in this study. Batch degradation of (14)C toluene was used to evaluate kinetic parameters for planktonic cells. The kinetic parameters determined for toluene degradation were: specific growth rate, mu(max) = 10.08 +/- 1.2/day; half-saturation constant, K(S) = 3.98 +/- 1.28 mg/L; substrate inhibition constant, K(I) = 42.78 +/- 3.87 mg/L. Biofilm cells, grown on ceramic rings in vapor phase bioreactors, were removed and suspended in batch cultures to calculate (14)C toluene degradation rates. Specific activities measured for planktonic and biofilm cells were similar based on toluene degrading cells and total biomass. Long-term toluene exposure reduced specific activities that were based on total biomass for both biofilm and planktonic cells. These results suggest that long-term toluene exposure caused a large portion of the biomass to become inactive, even though the biofilm was not substrate limited. Conversely, specific activities based on numbers of toluene-culturable cells were comparable for both biofilm and planktonically grown cultures. Planktonic cell kinetics are often used in bioreactor models to model substrate degradation and growth of bacteria in biofilms, a procedure we found to be appropriate for this organism. For superior bioreactor design, however, changes in cellular activity that occur during biofilm development should be investigated under conditions relevant to reactor operation before predictive models for bioreactor systems are developed. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 53: 535-546, 1997.     

Biodegradation by immobilized bacteria in an airlift-loop reactor-influence of biofilm diffusion limitation

Naphthalene-2-sulfonate was degraded by submerse growing Pseudomonads in a chemostat culture. The kinetic parameters for the Monod equation, including Pirts maintenance energy, were calculated from these experiments regarding naphthalene-2-sulfonate as substrate and oxygene as cosubstrate. By immobilizing the bacteria on sand particles, the degradation of naphthalene-2-sulfonate was carried out in a specialy designed three-phase airlift-loop reactor in a completely fluidized state. From these experiments, the influence of biofilm diffusion limitation on reaction kinetics and criteria for stable biofilm formation on sand particles were obtained.     

Nickel removal from nickel plating waste water using a biologically active moving-bed sand filter

Efficient removal of dissolved nickel was observed in a biologically active moving-bed `MERESAFIN' sand filter treating rinsing water from an electroless nickel plating plant. Although nickel is fully soluble in this waste water, its passage through the sand filter promoted rapid removal of approximately 1 mg Ni/l. The speciation of Ni in the waste water was modelled; the most probable precipitates forming under the conditions in the filter were predicted using PHREEQC. Analyses of the Ni-containing biosludge using chemical, electron microscopical and X-ray spectroscopic techniques confirmed crystallisation of nickel phosphate as arupite (Ni₃(PO₄)₂.8H₂O), together with hydroxyapatite within the bacterial biofilm on the filter sand grains. Biosorption contributed less than 1% of the overall sequestered nickel. Metabolising bacteria are essential for the process; the definitive role of specific components of the mixed population is undefined but the increase in pH promoted by metabolic activity of some microbial components is likely to promote nickel desolubilisation by others.     

Enhanced production of laccase in <Emphasis Type="Italic">Coriolopsis rigida</Emphasis> grown on barley bran in flask or expanded-bed bioreactor

In this work, the effect on laccase activity of adding xylidine, veratryl alcohol and copper sulphate to cultures of Coriolopsis rigida under submerged cultivation conditions have been investigated. The highest activities (approximately 6 × 10⁵ nkat/l) were obtained when the inducers copper sulphate (2 mM) and xylidine (10 mM) were added simultaneously. In addition, operating in the optimal conditions, it was possible to maintain the sustained production of laccase (around 3 × 10⁵ nkat/l) for successive repeated batch cultures in an expanded-bed laboratory scale bioreactor. On the other hand, in vitro phenol degradation by laccase obtained in the bioreactor was studied with/without an effective mediator 1-hydroxybenzotriazol (HBT). The presence of a radical mediator plays an important role inducing the degradation of phenol, and without mediator the polymerization of phenol was detected.