Column cellulose hydrolysis reactor: An efficient cellulose hydrolysis reactor with continuous cellulase recycling

Summary The use of a column cellulose hydrolysis reactor with continuous enzyme recycling was demonstrated by incorporating a continuous ultrafiltration apparatus at the effluent end of the column reactor. Using this setup, over 90% (w/v) cellulose hydrolysis was achieved, resulting in an average sugar concentration of 6.8% (w/v) in the effluent stream. The output of the system was 1.98 g of reducing sugar/l/h with a ratio of 87% (w/v) of the reducing sugars being monomeric sugars. Batch hydrolysis reactors were less effective, resulting in 57% (w/v) of the cellulose being hydrolyzed. The output of the batch reactor was 1.33 g of reducing sugar/l/h with similar product concentrations and percentage of monomeric sugars. The ratio of reducing sugar/filter paper unit of cellulase activity for the column method was 69.1 mg/U as compared to only 21.2 mg/U for the batch reactor.     

Evaluation of a upflow anaerobic sludge blanket reactor with partial recirculation of effluent used to treat wastewaters from pulp and paper plants

The main purpose of this study was to evaluate the performance of a UASB reactor treating diluted black liquor from a Kraft pulp mill, which simulates an unbleached Kraft plant wastewater, under different operational conditions, including partial recycling of the effluent. The reactor's performance was evaluated from the standpoint of COD, pH, volatile acid concentration, alkalinity, concentration of methane in the biogas, and microbiological examinations of the sludge. Without recirculation the reduction of the HRT from 36 to 30h did not significantly affect the average COD removal efficiency. The parameter displaying the greatest variation was the average concentration of effluent volatile acids, which increased by 16%. With recirculation the reduction of the HRT from 30 to 24h increased the average COD removal efficiency from 75% to 78%. In this case, the average effluent alkalinity also showed an increase. The use of partial recirculation of the effluent did not improve significantly the COD removal under the operational conditions tested in this work, but it was possible to operate the reactor with lower hydraulic retention time without disintegration of the granules.     

The influence of different substrate pH values on the performance of a downflow anaerobic fixed bed reactor treating a petrochemical effluent

Summary Neutralizing requirements for the anaerobic treatment of an acidic petrochemical effluent in a downflow anaerobic fixed bed reactor were examined. Neutralization (pH 6.0 with NaOH) of the effluent prior to digestion resulted in a Na⁺ concentration of over 3 g/l which was detrimental. Decreasing the Na⁺ concentration and subsequent replacement of NaOH by a mixture of Ca(OH)₂, NaOH and KOH resulted in an increase in reactor performance. The addition of different alkalines resulted in the best loading rates thusfar applied in the anaerobic treatment of this petrochemical effluent. During the final stages of this study, the effluent (pH 3.95) was treated at a loading rate of 10.37 kg COD/m³. d (HRT=1.35 d) with more than 94% fatty acid removal.     

Solubility and selective crystallization of lactose from solutions of its hydrolysis products glucose and galactose

A high degree of conversion is desired when lactose is hydrolyzed to glucose and galactose. This produces, however, a high concentration of galactose, which is inhibitory for the enzyme catalyst (beta-galactosidase). The inhibition can be reduced by limiting the conversion per pass over the enzyme (e.g. to ca. 50%), separating unconverted lactose from the reactor effluent, and recycling it to the reactor inlet. (This allows the overall conversion to be raised to ca. 80-90%). The solubilities of lactose, glucose, and galactose have been determined at various temperatures and for sugar mixtures having different concentrations and degrees of hydrolysis. Various cooling crystallizations have defined convenient and simple processes for the selective separation of lactose from its hydrolysis products.     

Anaerobic treatment of pre-hydrolysate liquor (PHL) from a rayon grade pulp mill: pilot and full-scale experience with UASB reactors

Pilot studies were carried out for the treatment of pre-hydrolysate liquor (PHL), a high strength effluent (COD: 70,000-80,000 mg/l) emanating from a rayon grade pulp mill using up-flow anaerobic sludge blanket reactor (UASB). Substrate inhibition was avoided with optimum COD feed of around 25,000 mg/l. This was achieved by diluting the PHL with a low strength effluent stream known as alkali back wash (ABW) available in the plant and also by partially recycling the reactor liquid outlet. An optimum organic loading rate (OLR) of 10 could be achieved with a COD reduction of 70-75%, a BOD reduction of 85-90% and a methane yield of 0.31-0.33 m3/kg of COD reduced. The pilot scale studies also revealed that addition of milk of lime (MOL) was essential for neutralization and buffering and DAP and urea to supplement the nutrients in the PHL. Based on the pilot studies, a full-scale high rate biomethanation plant was designed and erected for treating the PHL, which after some modification showed similar performance for COD, BOD reduction and methane yield.     

Effluent recirculation to improve perchlorate reduction in a fixed biofilm reactor

The effect of effluent recirculation on perchlorate reduction in a nominally plug-flow fixed biofilm reactor was studied in two cases: influent concentrations of 10 and 400 microg/L at low hydraulic loading rates (1.9 and 37.5 m(3)/m(2)/day without and with recirculation, respectively) and after a step increase in perchlorate concentration to 1,000 microg/L at the higher hydraulic loading rate (5 and 100 m(3)/m(2)/day without and with recirculation, respectively). Complete perchlorate reduction was sustained for influent concentrations of 400 and 10 microg/L in both flow regimes at the lower hydraulic loading rates. Reactor tracer profiles showed that biofilm diffusion had a more significant effect on mass transfer in the plug flow reactor compared with recirculation. The recirculation bioreactor acclimated more rapidly to increased hydraulic and perchlorate mass loading rates with significantly lower effluent perchlorate compared to the plug flow reactor: 16 microg/L versus 46 microg/L, respectively, although complete perchlorate removal was not achieved in either flow regime after 21 days acclimation to the higher loading. Total biofilm mass was more uniformly distributed in the recirculation reactor which may have contributed to better performance under increased perchlorate loading.     

Feasibility study of a pilot-scale sewage treatment system combining an up-flow anaerobic sludge blanket (UASB) and an aerated fixed bed (AFB) reactor at ambient temperature

A feasibility test of a 17 m3-pilot-scale sewage treatment system was carried out by continuous feeding of raw municipal sewage under ambient temperature conditions. The system consisted of a UASB and an aerated fixed bed reactor. Some of the effluent from the fixed bed reactor was returned to the UASB influent in order to provide a sulfate source. The total BOD of 148-162 mg l(-1) in the influent was reduced to a more desirable 11-25 mg l(-1) in the final effluent. The levels of methane-producing activity from acetate and H2/CO2 gas at 10 degrees C were only 2% and 0% of those at 35 degrees C, respectively. On the other hand, the sulfate-reducing activity levels of the UASB sludge were relatively high at 10 degrees C, for example, 18% for acetate and 9% for H2/CO2 gas, compared to the activity levels at 35 degrees C. Therefore, BOD oxidization by sulfate reduction in the UASB was greater than that by methane production under low temperature conditions. This sulfate-reducing activity tended to be proportional to the copy number of adenosine-5'-phosphosulfate (APS) reductase genes in DNA extracted from the sludge.     

Formation of metabolites during biodegradation of linear alkylbenzene sulfonate in an upflow anaerobic sludge bed reactor under thermophilic conditions.

Biodegradation of linear alkylbenzene sulfonate (LAS) was shown in an upflow anaerobic sludge blanket reactor under thermophilic conditions. The reactor was inoculated with granular biomass and fed with a synthetic medium and 3 micromol/L of a mixture of LAS with alkylchain length of 10 to 13 carbon atoms. The reactor was operated with a hydraulic retention time of 12 h with effluent recirculation in an effluent to influent ratio of 5 to 1. A sterile reactor operated in parallel revealed that sorption to sludge particles initially accounted for a major LAS removal. After 8 days of reactor operation, the removal of LAS in the reactor inoculated with active granular biomass exceeded the removal in the sterile reactor inoculated with sterile granular biomass. The effect of sorption ceased after 185 to 555 h depending on the LAS homologs. 40% of the LAS was biodegraded, and the removal rate was 0.5 x 10(-6) mol/h/mL granular biomass. Acidified effluent from the reactor was subjected to dichloromethane extraction followed by gas chromatography/mass spectrometry. Benzenesulfonic acid and benzaldehyde were detected in the reactor effluent from the reactor with active granular biomass but not in the sterile and unamended reactor effluent. Benzenesulfonic acid and benzaldehyde are the first identified degradation products in the anaerobic degradation of LAS.     

Integrated production of biogas and protein biomass from piggery manure

Summary An integrated process involving the production of biogas and the recovery of proteins by anaerobic digestion of piggery manure has been studied. the digestion, effected in a 616 L downflow stationary fixed film reactor, resulted in a biogas production rate of approximately 0.86 m³ per m³ reactor per day (35°C, hydraulic retention time 7.2 days). Treatment of the digested effluent by flocculation, decantation and/or sieving yielded a solid biomass with reduced coliform counts and a protein content of about 14%.     

Anaerobic treatment of distillery spent wash - a study on upflow anaerobic fixed film bioreactor

Anaerobic digestion of wastewater from a distillery industry having very high COD (1,10,000-1,90,000 mg/L) and BOD (50,000-60,000 mg/L) was studied in a continuously fed, up flow fixed film column reactor using different support materials such as charcoal, coconut coir and nylon fibers under varying hydraulic retention time and organic loading rates. The seed consortium was prepared by enrichment with distillery spent wash in a conventional type reactor having working capacity of 3 L and was used for charging the anaerobic column reactor. Amongst the various support materials studied the reactor having coconut coir could treat distillery spent wash at 8d hydraulic retention time with organic loading rate of 23.25 kg COD m(-3)d(-1) leading to 64% COD reduction with biogas production of 7.2 m3 m(-3)d(-1) having high methane yield without any pretreatment or neutralization of the distillery spent wash. This study indicates fixed film biomethanation of distillery spent wash using coconut coir as the support material appears to be a cost effective and promising technology for mitigating the problems caused by distillery effluent.     

Development of an immobilized cell reactor for the production of 1,3-propanediol by Citrobacter freundii

Citrobacter freundii DSM 30040 immobilized on modified polyurethane carrier particles PUR 90/16 was used for continuous glycerol fermentation in an anaerobic fixed bed reactor with effluent recycle and pH control (fixed bed loop reactor). The fermentor was run with buffered mineral medium under growth conditions resulting in the permanent renewal of active biomass. The effects of glycerol concentration in the feed, dilution rate (D), pH and temperature (T) were investigated to optimize the process. With 400 mm glycerol in the feed, pH 6.9, T = 36°C and D = 0.5 h^–1 the maximum productivity could be determined as 8.2 g/l per hour of 1,3-propanediol.     

Lipase-catalyzed selective synthesis of monolauroyl maltose using continuous stirred tank reactor

Monolauroyl maltose was synthesized by an immobilized lipase that catalyzed condensation of maltose and lauric acid in acetone using a batch reactor or a continuous stirred tank reactor. Mono- and di-lauroyl maltoses were identified by FT-IR, ¹H NMR, ¹³C NMR and MS. Monolauroyl maltose was selectively synthesized in a continuous stirred tank reactor and no diester was detected. The highest concentration of monolauroyl maltose at 28 mmol/l was obtained in 250 ml acetone when maltose was added at 4 g/d and the molar ratio of lauric acid to maltose was fixed at 4:1 at a flow rate of 0.15 ml/min for both influx and effluent without supplement of fresh molecular sieve.     

A new method of autotrophic denitrification with spent sulfidic caustic as substrate and alkalinity source

A new method based on sulfide utilizing autotrophic denitrification was adopted to remove nitrate from wastewater and to reuse spent sulfidic caustic containing high sulfide and alkalinity levels. The experiments were performed using a bench-scale upflow anoxic hybrid growth reactor (UAHGR) and an upflow anoxic suspended growth reactor (UASGR) to characterize the stoichiometric relationship between sulfur and nitrate in the process as well as the performance of the reactors. The level of nitrate removal from the UAHGR and UASGR were maintained at over 90% at a nitrate loading rate ranging from 0.15∼0.40 kgNO₃ ⁻/m³·d and no significant nitrite accumulation was observed in either reactor. Although the influent pH values were higher than the optimum range of autotrophic denitrification at 8.7∼10.1, the effluent pH was stable at 7.2∼7.9 due to the production of hydrogen ions during operation. The stoichiometric ratio of sulfate production to nitrate removal was 1.5∼2.1 mgSO₄ ²⁻/mgNO₃ ⁻ in both reactors. A comparison of the reactor performance revealed that the chemical parameters of the UAHGR operation corresponded to a plug flow like type reactor while the chemical parameters of the UASGR operation corresponded to a completely stirred tank reactor like type reactor. UAHGR did not require sludge recycling due to the packed media while UASGR required 300∼700% sludge recycling. Therefore, spent sulfidic caustic could be used in the sulfur utilizing autotrophic denitrification processes as substrate and alkalinity sources.     

Kinetics of a hollow fiber dehydrogenase reactor

A hollow fiber module was used as a reactor for conversion of ethanol to acetaldehyde in the presence of horse liver alcohol dehydrogenase as catalyst. Mass transport rates for NAD⁺, the overall acetaldehyde generation rate, catalyst effectiveness factors, and the overall order of the reaction with respect to NAD⁺ concentration were measured. A coupled-substrate reactor with continuous in situ regeneration of cofactor was also examined. Two substrates of opposite redox state were added simultaneously to the feed stream. NADH and acetaldehyde concentrations were monitored in the effluent stream. The cofactor recycle number, or ratio of moles of product to moles of NADH produced, exceeded 10,000 under certain conditions. While decreasing the NAD⁺ concentration in the feed stream decreased reactor productivity somewhat, it greatly enhanced the ratio of product formed per mole of NAD⁺ fed to the reactor. It is suggested that high cofactor costs in dehydrogenase reactors may be overcome with efficient in situ regeneration and secondary recovery and recycling of cofactor from the process stream.     

UASB/Flash aeration enable complete treatment of municipal wastewater for reuse

A simple, efficient and cost-effective method for municipal wastewater treatment is examined in this paper. The municipal wastewater is treated using an upflow anaerobic sludge bed (UASB) reactor followed by flash aeration (FA) as the post-treatment, without implementing aerobic biological processes. The UASB reactor was operated without recycle, at hydraulic retention time (HRT) of 8 h and achieved consistent removal of BOD, COD and TSS of 60-70% for more than 12 months. The effect of FA on UASB effluent post-treatment was studied at different HRT (15, 30 and 60 min) and dissolved oxygen (DO) concentrations (low DO = 1-2 mg/L and high DO = 5-6 mg/L). The optimum conditions for BOD, COD and sulfide removal were 30-60 min HRT and high DO concentration inside the FA tank. The final effluent after clarification was characterized by BOD and COD values of 28-35 and 50-58 mg/L, respectively. Sulfides were removed by more than 80%, but the fecal coliform only by ~2 log. The UASB followed by FA is a simple and efficient process for municipal wastewater treatment, except for fecal coliform, enabling water and nutrients recycling to agriculture.     

Bioremediation of selenite in oil refinery wastewater

Selenium-oxyanion-containing wastewater, with levels of selenite as high as 3690 g Se/l and very low levels of selenate, was treated in a laboratory-scale biological reactor system inoculated with the selenate-respiring bacterium Thauera selenatis. The wastewater contained selenite that had been removed from refinery effluent wastewater using iron-coprecipitation followed by selenite release to yield a more concentrated selenium-containing wastewater. The reactor system consisted of recycling sludge-blanket (500 ml; 200 g sand) and fluidized-bed reactors (500 ml; 150 g sand). The flow rate through the reactor system was 3.5 ml/min. The carbon source fed into the reactor was acetate (3 mM); nitrate was also present (3 mM). The selenium oxyanion levels in the wastewater were reduced by 95%. T. selenatis was the only selenate-reducing bacterium detected in the reactor system and it presumably reduced a portion of the selenate present in the water to selenite. The selenite present in the water, and that formed by selenate reduction, was reduced both by the Thauera and by a population of denitrifying bacteria also present in high numbers in the reactor system.     

Statistical determination of steady state condition in bioremediation tests

Linear regression and two newly developed statistical techniques were used to determine steady states in the dependent y-variables (effluent concentration or removal rate of pollutants) using correlation coefficients (r) for the relationship between the independent x-variables (reactor operating or treatment time) and the dependent y-variables. The statistical technique applied to chlorophenol bioremediation using a varying number of data points for linear regression analysis was more useful in determining a steady state for six general data patterns from bioremediation tests than the statistical technique using a fixed number of data points for linear regression analysis.     

Continuous adsorption and recovery of Cr(VI) in different types of reactors

This study reports the results of experiments on continuous adsorption and desorption of Cr(VI) ions by a chemically modified and polysulfone-immobilized biomass of the fungus Rhizopus nigricans. A fixed quantity of polymer-entrapped biomass beads corresponding to 2 g of dry biomass powder was employed in packed bed, fluidized bed, and stirred tank reactor for monitoring the continuous removal and recovery of Cr(VI) ions from aqueous solution and synthetic chrome plating effluent. Parameters such as flow rate (5, 10 and 15 mL/min), inlet concentration of Cr(VI) ions (50, 100, 150 and 250 mg/L) and the depth of biosorbent packing (22.8, 11.2 and 4.9 cm) were evaluated for the packed bed reactor. The breakthrough time and the adsorption rates in the packed bed column were found to decrease with increasing flow rate and higher Cr inlet concentrations and to increase with higher depths of sorbent packing. To have a comparative analysis of Cr adsorption efficiency in different types of reactors, the fluidized bed reactor and stirred tank reactor were operated using the same quantities of biosorbent material. For the fluidized bed reactor, Cr(VI) solution of 100 mg/L was pumped at 5 mL/min and fluidized by compressed air at a flow rate of 0.5 kg/cm.(2) The stirred tank reactor had a working volume of 200 mL capacity and the inlet/outlet flow rate was 5 mL/min. The maximum removal efficiency (mg Cr/g biomass) was obtained for the stirred tank reactor (159.26), followed by the fluidized reactor (153.04) and packed bed reactor (123.33). In comparison to the adsorption rate from pure chromate solution, approximately 16% reduction was monitored for synthetic chrome plating effluent in the packed bed. Continuous desorption of bound Cr ions from the reactors was effective with 0.01 N Na(2)CO(3) and nearly 80-94% recoveries have been obtained for all the reactors.     

Racemization of undesired enantiomers: Immobilization of mandelate racemase and application in a fixed bed reactor

Production of optically pure products can be based on simple unselective synthesis of racemic mixtures combined with a subsequent separation of the enantiomers; however, this approach suffers from a 50% yield limitation which can be overcome by racemization of the undesired enantiomer and recycling. Application of biocatalyst for the racemization steps offers an attractive option for high‐yield manufacturing of commercially valuable compounds. Our work focuses on exploiting the potential of racemization with immobilized mandelate racemase. Immobilization of crude mandelate racemase via covalent attachment was optimized for two supports: Eupergit^® CM and CNBr‐activated Sepharose 4 Fast Flow. To allow coupling of enzymatic reaction with enantioselective chromatography, a mobile phase composition compatible with both processes was used in enzymatic reactor. Kinetic parameters obtained analyzing experiments carried out in a batch reactor could be successfully used to predict fixed‐bed reactor performance. The applicability of the immobilized enzyme and the determined kinetic parameters were validated in transient experiments recording responses to pulse injections of R‐mandelic acid. The approach investigated can be used for futher design and optimization of high yield combined resolution processes. The characterized fixed‐bed enzymatic reactor can be integrated e.g. with chromatographic single‐ or multicolumn steps in various configurations.     

Highly sensitive enzyme thermistor determination of ADP and ATP by multiple recycling enzyme systems

An enzyme thermistor method for the determination of ADP and/or ATP with signal amplification by recycling procedures is described. Pyruvate kinase (PK) and hexokinase (HK) coimmobilised on aminopropyl-controlled pore glass were applied in a column reactor. Addition of an excess of phosphoenolpyruvate (PEP) and glucose leads to cofactor recycling and production of glucose-6-phosphate and pyruvate. In presence of PEP an amplification of the sensitivity up to 30 times was reached as compared with the HK-catalysed reaction alone. An additional signal amplification was accomplished by recycling the pyruvate leaving the first enzyme reactor in a second reactor containing L-lactate dehydrogenase, lactate oxidase and catalase. In the presence of NADH an overall amplification of the sensitivity for ATP or ADP up to 1700 times was found. The limits of detection were 6 × 10⁻⁵ M cofactor without recycling at all, 2 × 10⁻⁶M with recycling in the kinase bienzyme reactor and 1 × 10⁻⁸M with the dual recycling system.