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1.
A cyclone reactor for microbial fermentation processes was developed with high oxygen transfer capabilities. Three geometrically similar cyclone reactors with 0.5?l, 2.5?l and 15?l liquid volume, respectively, were characterized with respect to oxygen mass transfer, mixing time and residence time distribution. Semi-empirically correlations for prediction of oxygen mass transfer and mixing times were identified for scale-up of cyclone reactors. A volumetric oxygen mass transfer coefficient k L a of 1.0?s?1 (available oxygen transfer rate with air: 29?kg?m?3?h?1) was achieved with the cyclone reactor at a volumetric power input of 40?kW?m?3 and an aeration gas flow rate of 0.2?s?1. Continuous methanol controlled production of formate dehydrogenase (FDH) with Candida boidinii in a 15?l cyclone reactor resulted in more than 100% improvement in dry cell mass concentration (64.5?g?l?1) and in about 100% improvement in FDH space-time yield (300?U?l?1?h?1) compared to steady state results of a continuous stirred tank reactor.  相似文献   

2.
E. coli ATCC 11105 was cultivated in a 10-1 stirred tank reactor and in a 60-1 tower loop reactor in batch and continuous operation. By on-line measurements of O2 and CO2 concentrations in the outlet gas, pH, temperature, cell mass concentration X as well as dissolved O2 concentration along the tower in the broth, gas holdup, broth recirculation rate through the loop and by offline measurements of substrate concentration DOC and cell mass concentration along the tower, the maximum specific growth rate m , yield coefficients Y X/S. Y X/DOC and were evaluated in stirred tank and tower loop in batch and continuous cultures with and without motionless mixers in the tower and at different broth circulation rates through the loop. To control the accuracy of the measurements the C balance was calculated and 95% of the C content was covered.The biological parameters determined depend on the mode of operation as well as on the reactor used. Furthermore, they depend on the recirculation rate of the broth and built-ins in the tower. The unstructured cell and reactor models are unable to explain these differences. Obviously, structured cell and reactor models are needed. The cell mass concentration can be determined on line by NADH fluorescence in balanced growth, if the model parameters are determined under the same operational conditions in the same reactor.List of Symbols a, b empirical parameters in Eq. (1) - CPR kg/(m3 h) CO2 production rate - C kg/m3 concentration - D l/h dilution rate - DOC kg/m3 dissolved organic carbon - I net. fluorescence intensity - K S kg/m3 Monod constant - k L a l/h volumetric mass transfer coefficient - OTR kg/(m3 h) oxygen transfer rate - OUR kg/(m3 h) oxygen utilization rate - RQ = CPR/OUR respiratory quotient - S kg/m3 substrate concentration - t h,min, s time - t u min recirculation time - t M min mixing time - v m3/h volumetric flow rate through the loop - X kg/m3 (dry) cell mass concentration - Y X/S yield coefficient of cell mass with regard to the consumed substrate - Y X/DOC yield coefficient of the cell mass with regard to the consumed DOC - Y X/O yield coefficient of the cell mass with regard to the consumed oxygen - Z relative distance in the tower from the aerator with regard to the height of the aerated broth - l/h specific growth rate - m l/h maximum specific growth rate Indices f feed - e outlet  相似文献   

3.
A miniaturized reactor system with on‐line measurement of respiration rates by membrane inlet mass spectrometry was applied for the on‐line metabolic flux analysis at different phases of a 1.2 L batch culture of lysine‐producing Corynebacterium glutamicum. For this purpose, cells taken from the batch culture were transferred into the 12 mL mini reactor, and incubated for 15 min with [1‐18O]glucose. Quantification of oxygen uptake rate and CO2 mass isotopomer production rates in combination with a simple metabolic model allowed the estimation of the flux partitioning ratio between the pentose phosphate pathway and glycolysis during the process. The relative flux into the pentose pathway increased during growth, and reached maxima at 11 and 17 h cultivation time coinciding with maxima of the differential lysine yield. The developed system is a promising tool for determination of metabolic flux dynamics in industrially relevant batch and fed‐batch cultures.  相似文献   

4.
Methanolysis of sunflower oil catalyzed by immobilized Aspergillus niger mycelium was studied in a packed-bed reactor. The optimal cultivation parameters for A. niger were determined using full factorial and steepest ascent experimental designs. Sunflower oil, yeast extract and soybean meal were selected as the best carbon and nitrogen sources and were used in the subsequent experiments. Intracellular lipase activity and cell mass concentration were respectively 3.2 and 2.4 times greater and cultivation period decreased by 24 h compared with the initial medium. The optimum values of these most significant parameters were as follows: sunflower oil (13.2 g/L), yeast extract (6.2 g/L), soybean meal (7.4 g/L) and incubation period (72 h) at 30 °C. With A. niger biocatalyst grown in optimized conditions, the biodiesel fuel yield reached 23.1% after sixth pass of recycled reaction mixture through the reactor.  相似文献   

5.
This paper describes a novel bioscrubber concept for biological flue gas desulfurization, based on the recycling of a cell suspension of sulfite/sulfate reducing bacteria between a scrubber and a sulfite/sulfate reducing hydrogen fed bioreactor. Hydrogen metabolism in sulfite/sulfate reducing cell suspensions was investigated using batch activity tests and by operating a completely stirred tank reactor (CSTR). The maximum specific hydrogenotrophic sulfite/sulfate reduction rate increased with 10% and 300%, respectively, by crushing granular inoculum sludge and by cultivation of this sludge as cell suspension in a CSTR. Operation of a sulfite fed CSTR (hydraulic retention time 4 days; pH 7.0; sulfite loading rate 0.5–1.5 g SO 3 2- l-1 d-1) with hydrogen as electron donor showed that high (up to 1.6 g l-1) H2S concentrations can be obtained within 10 days of operation. H2S inhibition, however, limited the sulfite reducing capacity of the CSTR. Methane production by the cell suspension disappeared within 20 days reactor operation. The outcompetition of methanogens in excess of H2 can be attributed to CO2 limitation and/or to sulfite or sulfide toxicity. The use of cell suspensions opens perspectives for monolith or packed bed reactor configurations, which have a much lower pressure drop compared to air lift reactors, to supply H2 to sulfite/sulfate reducing bioreactors.  相似文献   

6.
Summary Cephalosporin C was produced by Cephalosporium acremonium in a 60 l airlift loop reactor on complex medium (with 30 kg/m3 peanut flour) in fed-batch operation. A final product concentration of 5 kg/m3 and a maximum productivity of 45 g/m3 h were attained. On-line analysis was used to determine ammonia, methionine, phosphate, reducing sugar and cephalosporin C by an autoanalyser, glucose by a flow injection analyser and cephalosporin C, penicillin N, deacetoxycephalosporin C, deacetylce-phalosporin C and methionine by HPLC. The volumetric productivity of the stirred tank reactor was higher than that of the airlift reactor because of differences in cell concentration. Specific productivities in relative to cell mass were similar in the two reactors. The substrate yield coefficient in the airlift reactor was twice that in the stirred tank reactor.Nomenclature E o2 efficiency of oxygen transfer with regard to the specific power input - K La volumetric mass transfer coefficient - OTR oxygen transfer rate - P power input - PR volumetric productivity of CPC - q a volumetric aeration rate/broth volume (vvm) - SPR specific productivity with regard to RNA - V L broth volume in reactor - z relative height of the aerated reactor  相似文献   

7.
The influences of geometric configuration, mycelial broth rheology and superficial gas velocity (Usg) were investigated with respect to the following hydrodynamic parameters: gas holdup (), oxygen transfer coefficient (KLa) and mixing time (tm). Increases in Usg and height of gas separator (Ht) caused an increase in and KLa, and a decrease in tm. Consequently, a diameter ratio (Dd/Dr) of 0.71 and Ht 0.20 m were found to be the best geometry and operation parameters to achieve high aeration and mixing efficiency for the high viscous broth system in the cultivation of filamentous fungi. An external airlift reactor (EALR) was developed and designed for the cultivation of filamentous fungi. The EALR with two spargers excels in reliability and high aeration and mass transfer coefficiency, resulting in a fast mycelial growth and high biomass productivity in the cultivation of the fungus Rhizopus oryzae.  相似文献   

8.
Protamylasse is a residual compound occurring during the industrial production of starch from potatoes. It contains a variety of nutrients and all necessary minerals and could be used as a carbon, nitrogen, and energy source for the growth of bacteria and also for cyanophycin (CGP) biosynthesis. Media containing protamylasse as the sole compound diluted only in water were therefore examined for their suitability in CGP production. Among various bacterial strains investigated in this study, a recombinant strain of Escherichia coli DH1 harboring plasmid pMa/c5-914::cphA6803, which carries the cyanophycin synthetase structural gene (cphA) from Synechocystis sp. strain PCC6803, was found to be most suitable. Various cultivation conditions for high CGP contents were first optimized in shake flask cultures. The optimized conditions were then successfully applied to 30- and 500-liter fermentation scales in stirred tank reactors. A maximum CGP content of 28% (wt/wt) CGP per cell dry matter was obtained in 6% (vol/vol) protamylasse medium at an initial pH of 7.0 within a cultivation period of only 24 h. The CGP contents obtained with this recombinant strain employing protamylasse medium were higher than those obtained with the same strain cultivated in mineral salts medium or in expensive commercial complex media such as Luria-Bertani or Terrific broth. It was shown that most amino acids present in the protamylasse medium were almost completely utilized by the cells during cultivation. Exceptions were alanine, tryptophan, tyrosine, and most interestingly, arginine. Furthermore, CGP was easily isolated from protamylasse-grown cells by applying the acid extraction method. The CGP exhibited a molecular mass of about 26 to 30 kDa and was composed of 50% (mol/mol) aspartate, 46% (mol/mol) arginine, and 4% (mol/mol) lysine. The use of cheap residual protamylasse could contribute in establishing an economically and also ecologically feasible process for the biotechnological production of CGP.  相似文献   

9.
Enzyme production with E. coli ATCC 11105, in a complex medium using phenylacetic acid as inducer is carried out in a stirred-tank reactor of 10 dm3 and an airlift tower-loop reactor of 60 dm3 with outer loop at a temperature of 27 °C. The optimum inducer concentration was 0.8 kg/m3, which was kept constant by fed-batch operation. The optimum of the relative dissolved O2-concentration with regard to saturation is below 10% in a stirred-tank reactor and at 35% in a tower-loop reactor. It was kept constant by parameter-adaptive control of the aeration rate. In a stirred-tank enzyme productivity is slightly higher than in a tower-loop reactor, and much higher than in a bubble column reactor.List of Symbols CPR kg/(m3 h) CO2-production rate - OTR kg/(m3 h) O2-transfer rate - OUR kg/(m3 h) O2-utilization rate - PAA phenylacetic acid (inducer) - RQ = CPR/OUR respiratory quotient - X kg/m3 cell mass concentration - m h–1 maximum specific growth rate  相似文献   

10.
Summary A novel membrane-stirrer reactor allowing bubble-free aeration was used for the cultivation of a cell suspension culture of Thalictrum rugosum. The immediate advantages were the absence of any foaming or flotation and the gentle mixing of the suspension by the tumbling movement of a coil or basket of membranes. In a 211 reactor under these conditions the suspensions were grown to high cell densities of 550 g fresh mass or 50 g dry mass/l. A pO2 of 30% was maintained up to 400 g/l without air bubble formation at the gas-exchange membranes. Provided that the physiological states of the initial suspensions were comparable, growth rates and alkaloid production of shake-flask and fermenter-grown cells were rather similar.Abbreviations NAA -naphthylacetic acid - 2,4-d 2,4-dichlorophenoxyacetic acid  相似文献   

11.
The kinetics of degradation of toluene from a model waste gas and of biomass formation were examined in a bioscrubber operated under different nutrient limitations with a mixed culture. The applicability of the kinetics of continuous cultivation of the mixed culture was examined for a special trickle-bed reactor with a periodically moved filter bed. The efficiency of toluene elimination of the bioscrubber was 50 to 57% and depended on the toluene mass transfer as evident from a constant productivity of 0.026 g dry cell weight/L . h over the dilution rate. Under potassium limitation the biomass productivity was reduced by 60% to 0.011 g dry cell weight/L . h at a dilution rate of 0.013/h. Conversely, at low dilution rates the specific toluene degradation rates increased. Excess biomass in a trickle-bed reactor causes reduction of interfacial area and mass transfer, and increase in pressure drop. To avoid these disadvantages, the trickle-bed was moved periodically and biomass was removed with outflowing medium. The concentration of steady state biomass fixed on polyamide beads decreased hyperbolically with the dilution rate. Also, the efficiency of toluene degradation decreased from 72 to 56% with increasing dilution rate while the productivity increased. Potassium limitation generally caused a reduction in biomass, productivity, and yield while the specific degradation increased with dilution rate. This allowed the application of the principles of the chemostat to the trickle-bed reactor described here, for toluene degradation from waste gases. (c) 1997 John Wiley & Sons, Inc. Biotechnol Bioeng 55: 686-692, 1997.  相似文献   

12.
Abstract

The ability of dried bacterial strain Bacillus sp. S14 to adsorb Malathion in a packed bed column reactor was studied. The effects of important design parameters such as bed height, flow rate and influent Malathion concentration on Malathion removal from an aqueous solution was studied using a packed bed column reactor. The optimised conditions for maximum Malathion removal were found to be: flow rate: 5 mL min-1, bed height: 6.0 cm and influent Malathion concentration: 25 mg L-1. The Adams-Bohart model, Wolborska model, Thomas model, Yoon and Nelson Model were employed to determine characteristic parameters such as N0 (saturation concentration, mg L-1), βo (external mass transfer coefficient, min-1), k Th(Thomas rate constant, mL min-1mg-1), q0 (maximum solid phase concentration of the solute, mg L-1), kYN (rate constant, min-1) and τ (time required for 50 % adsorbate breakthrough time, min) which are useful for process design. Data were fitted with Adams-Bohart model at lower region of (C/C0) values but more accurately fitted with Wolborska and Thomas model.  相似文献   

13.

Background

Among disposable bioreactor systems, cylindrical orbitally shaken bioreactors show important advantages. They provide a well-defined hydrodynamic flow combined with excellent mixing and oxygen transfer for mammalian and plant cell cultivations. Since there is no known universal correlation between the volumetric mass transfer coefficient for oxygen kLa and relevant operating parameters in such bioreactor systems, the aim of this current study is to experimentally determine a universal kLa correlation.

Results

A Respiration Activity Monitoring System (RAMOS) was used to measure kLa values in cylindrical disposable shaken bioreactors and Buckingham’s π-Theorem was applied to define a dimensionless equation for kLa. In this way, a scale- and volume-independent kLa correlation was developed and validated in bioreactors with volumes from 2 L to 200 L. The final correlation was used to calculate cultivation parameters at different scales to allow a sufficient oxygen supply of tobacco BY-2 cell suspension cultures.

Conclusion

The resulting equation can be universally applied to calculate the mass transfer coefficient for any of seven relevant cultivation parameters such as the reactor diameter, the shaking frequency, the filling volume, the viscosity, the oxygen diffusion coefficient, the gravitational acceleration or the shaking diameter within an accuracy range of +/? 30%. To our knowledge, this is the first kLa correlation that has been defined and validated for the cited bioreactor system on a bench-to-pilot scale.
  相似文献   

14.
An industrial scale reactor concept for continuous cultivation of immobilized animal cells (e.g. hybridoma cells) in a radial-flow fixed bed is presented, where low molecular weight metabolites are removed via dialysis membrane and high molecular products (e.g. monoclonal antibodies) are enriched. In a new nutrient-split feeding strategy concentrated medium is fed directly to the fixed bed unit, whereas a buffer solution is used as dialysis fluid. This feeding strategy was investigated in a laboratory scale reactor with hybridoma cells for production of monoclonal antibodies. A steady state monoclonal antibody concentration of 478 mg l-1 was reached, appr. 15 times more compared to the concentration reached in chemostat cultures with suspended cells. Glucose and glutamine were used up to 98%. The experiments were described successfully with a kinetic model for immobilized growing cells. Conclusions were drawn for scale-up and design of the large scale system.Abbreviations: cGlc – glucose concentration, mmol l-1; cGln – glutamine concentration, mmol l-1; cAmm – ammonia concentration, mmol l-1; cLac – lactate concentration, mmol l-1; cMAb – MAb concentration, mg l-1; D – dilution rate, d-1; Di – dilution rate in the inner chamber of the membrane dialysis reactor, d-1; D0 – dilution rate in the outer chamber of the membrane dialysis reactor, d-1; q*FB,Glc – volume specific glucose uptake rate related to the fixed bed volume, mmol lFB -1 h-1; q*FB,Gln – volume specific glutamine uptake rate related to the fixed bed volume, mmol lFB -1 h-1.  相似文献   

15.
The impact of mass transfer on productivity can become a crucial aspect in the fermentative production of bulk chemicals. For highly aerobic bioprocesses the oxygen transfer rate (OTR) and productivity are coupled. The achievable space time yields can often be correlated to the mass transfer performance of the respective bioreactor. The oxygen mass transfer capability of a jet aerated loop reactor is discussed in terms of the volumetric oxygen mass transfer coefficient kLa [h?1] and the energetic oxygen transfer efficiency E [kgO2 kW?1 h?1]. The jet aerated loop reactor (JLR) is compared to the frequently deployed aerated stirred tank reactor. In jet aerated reactors high local power densities in the mixing zone allow higher mass transfer rates, compared to aerated stirred tank reactors. When both reactors are operated at identical volumetric power input and aeration rates, local kLa values up to 1.5 times higher are possible with the JLR. High dispersion efficiencies in the JLR can be maintained even if the nozzle is supplied with pressurized gas. For increased oxygen demands (above 120 mmol L?1 h?1) improved energetic oxygen transfer efficiencies of up to 100 % were found for a JLR compared to an aerated stirred tank reactor operating with Rushton turbines.  相似文献   

16.
A new alkaline keratinase, which could kill Meloidogyne incognita (a root-knot nematode) was separated and purified from Bacillus sp. 50-3 in this study. The solid ammonium sulfate was selected to precipitate the enzyme and its proper adding mass was also determined. After solid ammonium sulfate precipitation and liquid chromatography on DEAE-Sephadex-A50 column, there was 17.7-fold purification with a yield of 46.5%, as determined by azokeratin as substrate. The purification effect was determined through SDS-PAGE and the molecular weight of the enzyme was found to be 27,423 Da by the MALDI-TOF-MS. When the second-stage juveniles of Meloidogyne incognita were exposed to 50 μg/ml of keratinase solution, 98.5% of Meloidogyne incognita mortality rates were obtained compared to control after 24 h. Its simple purification step and high yield from the cheap medium affords this keratinase great biotechnological potential, especially in controlling root-knot nematodes such as Meloidogyne incognita. To the best of our knowledge, this study is the first report that uses keratinase as a pesticide.  相似文献   

17.
A new bioreactor system has been developed for in vivo NMR spectroscopy of microorganisms under defined physiological conditions. This cyclone reactor with an integrated NMR flow cell is continuously operated in the magnet of a 400-MHz wide-bore NMR spectrometer system. The residence times of medium and cells are decoupled by a circulation-integrated cross-flow microfiltration module to achieve higher cell densities as compared to continuous fermentations without cell retention (increase in cell density up to a factor of 10 in steady state). Volumetric mass transfer coefficients k(L)a of more than 1.0 s(-1) are possible in the membrane cyclone reactor, ensuring adequate oxygen supply [oxygen transfer rate >15,000 mg O(2) .(L h)(-1)] of high cell densities. With the aid of the membrane cyclone reactor we were able to show, using continuous in vivo (31)P NMR spectroscopy of anaerobic glucose fermentation by Zymomonas mobilis, that the NMR signal intensity was directly proportional to the cell concentration in the reactor. The concentration profiles of intracellular inorganic phosphate, NAD(H), NDP, NTP, UDP-sugar, a cyclic pyrophosphate, two sugar phosphate pools, and extracellular inorganic phosphate were recorded after a shift from one steady state to another. The intracellular cyclic pyrophosphate had not been detected before in in vitro measurements of Zymomonas mobilis extracts due to the high instability of this compound. Using continuous in vivo (13)C NMR spectroscopy of aerobic glucose utilization by Corynebacterium glutamicum at a density of 25 g(cell dry weight) . L(-1), the membrane cyclone reactor served to measure the different dynamics of labeling in the carbon atoms of L-lactate, L-glutamate, succinate, and L-lysine with a time resolution of 10 min after impressing a [1-(13)C]-glucose pulse.  相似文献   

18.
Polysialic acid (polySia), consisting of α‐(2,8)‐linked N‐acetylneuraminic acid monomers plays a crucial role in many biological processes. This study presents a novel process for the production of endogenous polySia using Escherichia coli K1 in a disposable bag reactor with wave‐induced mixing. Disposable bag reactors provide easy and fast production in terms of regulatory requirements as GMP, flexibility, and can easily be adjusted to larger production capacities not only by scale up but also by parallelization. Due to the poor oxygen transfer rate compared to a stirred tank reactor, pure oxygen was added during the cultivation to avoid oxygen limitation. During the exponential growth phase the growth rate was 0.61 h?1. Investigation of stress‐related product release from the cell surface showed no significant differences between the disposable bag reactor with wave‐induced mixing and the stirred tank reactor. After batch cultivation a cell dry weight of 6.8 g L?1 and a polySia concentration of 245 mg L?1 were reached. The total protein concentration in the supernatant was 132 mg L?1. After efficient and time‐saving downstream processing characterization of the final product showed a protein content of below 0.04 mgprotein/gpolySia and a maximal chain length of ~90 degree of polymerization.  相似文献   

19.
A specially designed model reactor based on a 42‐L laboratory fermentor was equipped with six stirrers (Rushton turbines) and five cylindrical disks. In this model reactor, the mixing time, Θ90, turned out to be 13 times longer compared with the 42‐L standard laboratory fermentor fitted with two Rushton turbines and four wall‐fixed longitudinal baffles. To prove the suitability of the model reactor for scaledown studies of mixing‐time‐dependent processes, parallel exponential fed‐batch cultivations were carried out with the leucine‐auxotrophic strain, Corynebacterium glutamicum DSM 5715, serving as a microbial test system. L‐ Leucine, the process‐limiting substrate, was fed onto the liquid surface of both reactors. Cultivations were conducted using the same inoculum material and equal oxygen supply. The model reactor showed reduced sugar consumption (−14%), reduced ammonium consumption (−19%), and reduced biomass formation (−7%), which resulted in a decrease in L ‐lysine formation (−12%). These findings were reflected in less specific enzyme activity, which was determined for citrate synthase (CS), phosphoenolpyruvate carboxylase (PEP‐C), and aspartate kinase (AK). The reduced specific activity of CS correlated with lower CO2 evolution (−36%) during cultivation. The model reactor represents a valuable tool to simulate the conditions of poor mixing and inhomogeneous substrate distribution in bioreactors of industrial scale. © 1999 John Wiley & Sons, Inc. Biotechnol Bioeng 64: 599–606, 1999.  相似文献   

20.
Summary The effect of the periodic variation of the partial pressure of oxygen in the aeration gas on biomass concentrations, ethanol conversion, yield and productivity during continuous cultivations of the yeast Candida utilis in a multistage tower fermentor was studied. The results were compared with those obtained under aeration conditions with a constant PO 2 in the aeration gas. The results demonstrated that, with the optimum PO 2 in the aeration gas, the aeration procedure with the periodic variation of PO 2 in the gas phase permitted achievement of the same process parameters as those under constant PO 2. Using this new aeration procedure, the consumption of pure oxygen can be lowered by 55% to 60%. In addition, the significance of the induction effect of PO 2 on growth characteristics in the individual stages of the fermentor was proved.Symbols Ac Concentration of acetic acid (g/l) - i Number of stage - PO 2 Partial pressure of oxygen in the aeration gas (torr) - PR Productivity of the fermentor (g cell dwt/l/h) - SR Ethanol concentration in the feed (g/l) - S Ethanol concentration in the cultivation broth (g/l) - t Time of continuous cultivation (h) - X Cell dry weight concentration (g/l) - (YX/S)W Yield of cell dry weight from ethanol for the whole fermentor (g cell dwt/g ethanol) - Concentration interval in which parameters varied during the long-term cultivation at constant constant PO 2=263.5 torr in the aeration gas - 1 Concentration interval in which parameters varied during the long-term cultivation before the increase of PO 2 in the aeration gas - 2 Concentration interval in which parameters varied during the long-term cultivation immediately after the decrtease of PO 2 in the aeration gas - 3 Concentration interval in which parameters varied during the long-term cultivation about 24 h after the decrease of PO 2 in the aeration gas - 4 Concentration interval in which parameters varied during the long-term cultivation about 48 h after the decrease of PO 2 in the aeration gas  相似文献   

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