AnSBBR Applied to the Treatment of Metalworking Fluid Wastewater: Effect of Organic and Shock Load

An investigation was performed regarding the application of a mechanically stirred anaerobic sequencing batch biofilm reactor containing immobilized biomass on inert polyurethane foam (AnSBBR) to the treatment of soluble metalworking fluids to remove organic matter and produce methane. The effect of increasing organic matter and reactor fill time, as well as shock load, on reactor stability and efficiency have been analyzed. The 5-L AnSBBR was operated at 30?°C in 8-h cycles, agitation of 400 rpm, and treated 2.0 L effluent per cycle. Organic matter was increased by increasing the influent concentration (500, 1,000, 2,000, and 3,000 mg chemical oxygen demand (COD)/L). Fill times investigated were in the batch mode (fill time 10 min) and fed-batch followed by batch (fill time 4 h). In the batch mode, organic matter removal efficiencies were 87%, 86%, and 80% for influent concentrations of 500, 1,000, and 2,000 mgCOD/L (1.50, 3.12, and 6.08 gCOD/L.d), respectively. At 3,000 mgCOD/L (9.38 gCOD/L.d), operational stability could not be achieved. The reactor managed to maintain stability when a shock load twice as high the feed concentration was applied, evidencing the robustness of the reactor to potential concentration variations in the wastewater being treated. Increasing the fill time to 4 h did not improve removal efficiency, which was 72% for 2,000 mgCOD/L. Thus, gradual feeding did not improve organic matter removal. The concentration of methane formed at 6.08 gCOD/L was 5.20 mmolCH₄, which corresponded to 78% of the biogas composition. The behavior of the reactor during batch and fed-batch feeding could be explained by a kinetic model that considers organic matter consumption, production, and consumption of total volatile acids and methane production.     

AnSBBR Applied to a Personal Care Industry Wastewater Treatment: Effects of Fill Time,Volume Treated Per Cycle,and Organic Load

A study was performed regarding the effect of the relation between fill time, volume treated per cycle, and influent concentration at different applied organic loadings on the stability and efficiency of an anaerobic sequencing batch reactor containing immobilized biomass on polyurethane foam with recirculation of the liquid phase (AnSBBR) applied to the treatment of wastewater from a personal care industry. Total cycle length of the reactor was 8 h (480 min). Fill times were 10 min in the batch operation, 4 h in the fed-batch operation, and a 10-min batch followed by a 4-h fed batch in the mixed operation. Settling time was not necessary since the biomass was immobilized and decant time was 10 min. Volume of liquid medium in the reactor was 2.5 L, whereas volume treated per cycle ranged from 0.88 to 2.5 L in accordance with fill time. Influent concentration varied from 300 to 1,425 mg COD/L, resulting in an applied volumetric organic load of 0.9 and 1.5 g COD/L.d. Recirculation flow rate was 20 L/h, and the reactor was maintained at 30 °C. Values of organic matter removal efficiency of filtered effluent samples were below 71% in the batch operations and above 74% in the operations of fed batch followed by batch. Feeding wastewater during part of the operational cycle was beneficial to the system, as it resulted in indirect control over the conversion of substrate into intermediates that would negatively interfere with the biochemical reactions regarding the degradation of organic matter. As a result, the average substrate consumption increased, leading to higher organic removal efficiencies in the fed-batch operations.     

Feasibility of treating swine manure in an anaerobic sequencing batch biofilm reactor with mechanical stirring

Anaerobic sequencing batch reactors containing granular or flocculent biomass have been employed successfully in the treatment of piggery wastewater. However, the studies in which these reactors were employed did not focus specifically on accelerating the hydrolysis step, even though the degradation of this chemical oxygen demand (COD) fraction is likely to be the limiting step in many investigations of this type of wastewater. The mechanically stirred anaerobic sequencing batch biofilm reactor offers an alternative for hastening the hydrolysis step, because mechanical agitation can help to speed up the reduction of particle sizes in the fraction of particulate organic matter. In the present study, a 4.5-L reactor was operated at 30°C, with biomass immobilized on cubic polyurethane foam matrices (1 cm of side) and mechanical stirring provided by three flat-blade turbines (6 cm) at agitation rates varying from 0 to 500 rpm. The reactor was operated to treat diluted swine waste, and mechanical stirring efficiently improved degradation of the suspended COD. The operational data indicate that the reactor remained stable during the testing period. After 2 h of operation at 500 rpm, the suspended COD decreased by about 65% (from 1500 to 380 mg/L). Apparent kinetic constants were also calculated by modified first-order expressions.     

AnSBBR Applied to Organic Matter and Sulfate Removal: Interaction Effect Between Feed Strategy and Cod/Sulfate Ratio

A mechanically stirred anaerobic sequencing batch reactor containing anaerobic biomass immobilized on polyurethane foam cubes, treating low-strength synthetic wastewater (500 mg COD L^?1), was operated under different operational conditions to assess the removal of organic matter and sulfate. These conditions were related to fill time, defined by the following feed strategies: batch mode of 10 min, fed-batch mode of 3 h and fed-batch mode of 6 h, and COD/[SO₄ ^2?] ratios of 1.34, 0.67, and 0.34 defined by organic matter concentration of 500 mg COD L^?1 and sulfate concentrations of 373, 746, and 1,493 mg SO₄ ^2? L^?1 in the influent. Thus, nine assays were performed to investigate the influence of each of these parameters, as well as the interaction effect, on the performance of the system. The reactor operated with agitation of 400 rpm, total volume of 4.0 L, and treated 2.0 L synthetic wastewater in 8-h cycles at 30?±?1°C. During all assays, the reactor showed operational stability in relation to the monitored variables such as COD, sulfate, sulfide, sulfite, volatile acids, bicarbonate alkalinity, and solids, thus demonstrating the potential to apply this technology to the combined removal of organic matter and sulfate. In general, the results showed that the 3-h fed-batch operation with a COD/[SO₄ ^2?] ratio of 0.34 presented the best conditions for organic matter removal (89%). The best efficiency for sulfate removal (71%) was accomplished during the assay with a COD/[SO₄ ^2?] ratio of 1.34 and a fill time of 6 h. It was also observed that as fill time and sulfate concentration in the influent increased, the ratio between removed sulfate load and removed organic load also increased. However, it should be pointed out that the aim of this study was not to optimize the removal of organic matter and sulfate, but rather to analyze the behavior of the reactor during the different feed strategies and applied COD/[SO₄ ^2?] ratios, and mainly to analyze the interaction effect, an aspect that has not yet been explored in the literature for batch reactors.     

ASBR Applied to the Treatment of Biodiesel Production Effluent: Effect of Organic Load and Fill Time on Performance and Methane Production

The effect of organic matter and fill time on anaerobic sequencing batch reactor (5 L, 30°C, 8-h cycles, 50 rpm) efficiency has been analyzed. Organic matter was increased by the influent concentration. Fill times investigated were in the batch mode and fed-batch followed by batch. In the batch mode organic matter removal were 93%, 81%, and 66% for influent concentration of 500, 1,000, and 2,000 mgCOD/L (0.6, 1.29, and 2.44 gCOD/L.d), respectively. At 3,000 mgCOD/L (3.82 gCOD/L.d) operational stability could not be achieved. Removal efficiency was improved by increasing the fill time, and was 85% for the 1,000 mgCOD/L condition and fill times of 2 and 4 h, and 80 and 77% for the 2,000 mgCOD/L condition and fill times of 2 and 4 h, respectively. Hence, gradual feeding seemed to improve and to smooth the profiles of organic matter and volatile acids along the cycle with 78 to 96 NmLCH₄/gCOD.     

Effect of Substrate Concentration on Dark Fermentation Hydrogen Production Using an Anaerobic Fluidized Bed Reactor

The effect of substrate (glucose) concentration on the stability and yield of a continuous fermentative process that produces hydrogen was studied. Four anaerobic fluidized bed reactors (AFBRs) were operated with a hydraulic retention time (HRT) from 1 to 8 h and an influent glucose concentration from 2 to 25 g L⁻¹. The reactors were inoculated with thermally pre-treated anaerobic sludge and operated at a temperature of 30 °C with an influent pH around 5.5 and an effluent pH of about 3.5. The AFBRs with a HRT of 2 h and a feed strength of 2, 4, and 10 g L⁻¹ showed satisfactory H₂ production performance, but the reactor fed with 25 g L⁻¹ of glucose did not. The highest hydrogen yield value was obtained in the reactor with a glucose concentration of 2 g L⁻¹ when it was operated at a HRT of 2 h. The maximum hydrogen production rate value was achieved in the reactor with a HRT of 1 h and a feed strength of 10 g L⁻¹. The AFBRs operated with glucose concentrations of 2 and 4 g L⁻¹ produced greater amounts of acetic and butyric acids, while AFBRs with higher glucose concentrations produced a greater amount of solvents.     

Analysis of performance of an anaerobic sequencing batch reactor submitted to increasing organic load with different influent concentrations and cycle lengths

The performance of an anaerobic sequencing batch reactor (ASBR) was assessed when submitted to increasing organic load with different influent concentrations and cycle lengths. The 5-L mechanically stirred (75 rpm) ASBR contained 2 L of granular biomass and treated 2 L of synthetic wastewater per cycle. Volumetric organic loads (VOLs) from 0.66 to 2.88 g of chemical oxygen demand (COD)/(L x d) were applied by using influent concentrations from 550 to 3,600 mg of COD/L in 8- and 12-h cycles. Reactor stability was maintained for VOLs from 0.66 to 2.36 g of COD/(L x d), with organic matter removal efficiencies for filtered samples (epsilonF) between 84 and 88%. For VOLs from 0.78 to 2.36 g of COD/(L x d) at an influent concentration of 2,000 mg of COD/L, when cycle length was reduced from 12 to 8 h, epsilonF did not vary, yet showed a very distinct behavior from the other conditions. In addition, two operation strategies were studied for VOLs with approximately similar values of 2.36 and 2.08 g of COD/(L x d). One involved operation with an influent concentration of 2,000 mg of COD/L and an 8-h cycle, whereas the other involved an influent concentration of 2,600 mg of COD/L and a 12-h cycle. Only the former resulted in system stability and efficiency. These results indicate that besides organic load, influent concentration and cycle length play a significant role in ASBR systems.     

Effect of Feed Strategy on Methane Production and Performance of an AnSBBR Treating Effluent from Biodiesel Production

The aim of this work was to investigate the effect of different feeding times (2, 4 and 6 h) and applied volumetric organic loads (4.5, 6.0 and 7.5 gCOD L⁻¹ day⁻¹) on the performance of an anaerobic sequencing batch biofilm reactor (AnSBBR) treating effluent from biodiesel production. Polyurethane foam cubes were used as inert support in the reactor, and mixing was accomplished by recirculating the liquid phase. The effect of feeding time on reactor performance showed to be more pronounced at higher values of applied volumetric organic loads (AVOLs). Highest organic material removal efficiencies achieved at AVOL of 4.5 gCOD L⁻¹ day⁻¹ were 87 % at 4-h feeding against 84 % at 2-h and 6-h feeding. At AVOL of 6.0 gCOD L⁻¹ day⁻¹, highest organic material removal efficiencies achieved with 4-h and 6-h feeding were 84 %, against 71 % at 2-h feeding. At AVOL of 7.5 gCOD L⁻¹ day⁻¹, organic material removal efficiency achieved with 4-h feeding was 77 %. Hence, longer feeding times favored minimization of total volatile acids concentration during the cycle as well as in the effluent, guaranteeing process stability and safety.     

The Effect of Biomass Immobilization Support Material and Bed Porosity on Hydrogen Production in an Upflow Anaerobic Packed-Bed Bioreactor

The aim of this study was to investigate the effect of the support material used for biomass attachment and bed porosity on the potential generation of hydrogen gas in an anaerobic bioreactor treating low-strength wastewater. For this purpose, an upflow anaerobic packed-bed (UAPB) reactor fed with sucrose-based synthetic wastewater was used. Three reactors with various support materials (expanded clay, vegetal coal, and low-density polyethylene) were operated for hydraulic retention time (HRT) of 0.5 and 2 h. Based on the results obtained, three further reactors were operated with low-density polyethylene as a material support using various bed porosities (91, 75, and 50 %) for an HRT of 0.5 h. The UAPB reactor was found to be a feasible technology for hydrogen production, reaching a maximum substrate-based hydrogen yield of 7 mol H₂ mol^?1 sucrose for an HRT of 0.5 h. The type of support material used did not affect hydrogen production or the microbial population inside the reactor. Increasing the bed porosity to 91 % provided a continuous and cyclic production of hydrogen, whereas the lower bed porosities resulted in a reduced time of hydrogen production due to biomass accumulation, which resulted in a decreasing working volume.     

Pretreatment of Synthetic Dairy Wastewater Using the Sophorolipid-Producing Yeast <Emphasis Type="Italic">Candida bombicola</Emphasis>

The presence of high strength fats and oils in dairy industry wastewaters poses serious challenges for biological treatment systems, and, therefore, its pretreatment is necessary in order to remove them. In the present study, synthetic dairy wastewater prepared in the laboratory was pretreated using the sophorolipid-producing yeast Candida bombicola in a laboratory-scale bioreactor under batch, fed-batch, and continuous modes of operation. To support the yeast growth, the wastewater was supplemented with sugarcane molasses (1% w/v) and yeast extract (0.1% w/v). Results from the batch operated fermentor revealed complete utilization of fats present in the wastewater within 96 h with more than 93% COD removal efficiency. The yeast was, however, able to pretreat the wastewater more quickly and efficiently under fed-batch mode of operation than under batch operated condition in the same fermentor. Continuous experiments were carried out with a wastewater retention time of 28 h in the reactor; results showed very good performance of the system in complete utilization of fats and COD removal efficiency of more than 90%. The study proved the excellent potential of the biosurfactant-producing yeast in pretreating high-fat- and oil-containing dairy industry wastewater.     

Sequential anaerobic/aerobic treatment of dye-containing wastewaters: colour and COD removals, and ecotoxicity tests

Colour and COD removals of the azo dyes Congo Red (CR) and Reactive Black 5 (RB5) were individually evaluated in a sequential anaerobic/aerobic treatment system. Additionally, dye toxicity was assessed by using acute ecotoxicity tests with Daphnia magna as the indicator-organism. The anaerobic reactor was operated at approximately 27 °C and with hydraulic retention times of 12 and 24 h. The aerobic reactor was operated in batch mode with a total cycle of 24 h. During anaerobic step, high colour removals were obtained, 96.3% for CR (400 mg/L) and 75% for RB5 (200 mg/L). During the aerobic phase, COD effluent was considerably reduced, with an average removal efficiency of 52% for CR and 85% for RB5, which resulted in an overall COD removal of 88% for both dyes. Ecotoxicity tests with CR revealed that the anaerobic effluent presented a higher toxicity compared with the influent, and an aerobic post-treatment was not efficient in reducing toxicity. However, the results with RB5 showed that both anaerobic and aerobic steps could decrease dye toxicity, especially the aerobic phase, which removed completely the toxicity in D. magna. Therefore, the anaerobic/aerobic treatment is not always effective in detoxifying dye-containing wastewaters, sometimes even increasing dye toxicity.     

Efficient monooleoyl glycerol synthesis employing hybrid ultrasonic-infrared-wave promoted reactor: Concurrent catalytic and noncatalytic esterification kinetics

For the first time, intensification of monooleoyl glycerol (MOG) synthesis has been investigated in an ultrasonic-infrared-wave (USIRW) promoted batch reactor. Esterification of octadecanoic acid (ODA) with glycerol (Gl) has been conducted [using Amberlyst 36 wet catalyst] in three different reactors, namely traditional batch reactor (TBR), infrared wave promoted batch reactor (IRWPBR), and USIRW-promoted batch reactor (USIRWPBR) to assess the relative efficacy. The energy-efficient USIRWPBR remarkably intensifies the ODA-Gl esterification as manifested through superior ODA conversion (92.5 ± 1.25%) compared to that achieved in IRWPBR (79.8 ± 1.2%) and TBR (36.39 ± 1.25%). The most favorable reaction condition for optimum ODA conversion and maximum MOG yield was identified through statistical optimization over a selected parametric range, namely 3-5 Gl/ODA mole ratio, 0.004-0.006 g/mL Amberlyst 36 catalyst concentration, 300-700 rpm impeller speed, and 333-353 K reaction temperature. The present study also reports the formulation and validation of an innovative reaction kinetics, that is, concurrent noncatalytic and heterogeneously catalyzed (CNCHC) reaction mechanism in addition to the conventional heterogeneous kinetic models (LH and Eley-Rideal mechanisms). Under combined USIRW, the CNCHC esterification mechanism could best describe ODA-Gl esterification (R² = 0.98) compared to LH (R² = 0.97) and Eley-Rideal (R² = 0.88) mechanisms. The optimal product (MOG) was characterized by differential scanning calorimetry and thermogravimetric analysis to assess its crystallization property and thermal stability for possible application as plasticizer/fuel additives.     

Biomethane Production in an AnSBBR Treating Wastewater from Biohydrogen Process

An anaerobic sequencing batch reactor containing immobilized biomass (AnSBBR) was used to produce biomethane by treating the effluent from another AnSBBR used to produce biohydrogen from glucose- (AR-EPHG) and sucrose-based (AR-EPHS) wastewater. In addition, biomethane was also produced from sucrose-based synthetic wastewater (AR-S) in a single AnSBBR to compare the performance of biomethane production in two steps (acidogenic and methanogenic) in relation to a one-step operation. The system was operated at 30 °C and at a fixed stirring rate of 300 rpm. For AR-EPHS treatment, concentrations were 1,000, 2,000, 3,000, and 4,000 mg chemical oxygen demand?(COD)?L^?1 and cycle lengths were 6 and 8 h. The applied volumetric organic loads were 2.15, 4.74, 5.44, and 8.22 g COD L^?1 day^?1. For AR-EPHG treatment, concentration of 4,000 mg COD L^?1 and 4-h cycle length (7.21 g COD L^?1 day^?1) were used. For AR-S treatment, concentration was 4,000 mg COD L^?1 day^?1 and cycle lengths were 8 (7.04 g COD L^?1 day^?1) and 12 h (4.76 g COD L^?1 day^?1). The condition of 8.22 g COD L^?1 day^?1 (AR-EPHS) showed the best performance with respect to the following parameters: applied volumetric organic load of 7.56 g COD L^?1 day^?1, yield between produced methane and removed organic material of 0.016 mol CH₄?g COD^?1, CH₄ content in the produced biogas of 85 %, and molar methane productivity of 127.9 mol CH₄?m^?3 day^?1. In addition, a kinetic study of the process confirmed the trend that, depending on the biodegradability characteristics of the wastewaters used, the two-step treatment (acidogenic for biohydrogen production and methanogenic for biomethane production) has potential advantages over the single-step process.     

The Effect of pH on Continuous Biohydrogen Production from Swine Wastewater Supplemented with Glucose

The effect of pH on hydrogen production from liquid swine manure supplemented with glucose by a mixed culture of fermentative bacteria in an anaerobic sequencing batch reactor was evaluated in this study. At 37 ± 1 °C, five pH values ranging from 4.7 to 5.9 at an increment of 0.3 were tested at a hydraulic retention time (HRT) of 16 h. The results showed that at this HRT, the optimal pH for hydrogen production was 5.0, under which the biogas comprised 33.57 ± 5.65% of hydrogen with a production rate of 8.88 ± 2.94 L-H₂/day and a yield of 1.48 ± 0.49 L-H₂/L liquid swine manure. The highest biomass concentration, highest butyric acid to acetic acid ratio, lowest propionic acid concentration, and the best stability were all found at pH 5.0, while the highest CH₄ productivity was found at pH 5.9. For efficient hydrogen production, oxygen content should be controlled under 2%, beyond which an inverse linear relationship (R ² = 0.986) was observed.     

Variable Volume Fed-Batch Fermentation for Nisin Production by <Emphasis Type="Italic">Lactococcus lactis</Emphasis> subsp<Emphasis Type="Italic">. lactis W28</Emphasis>

A feeding technology that was suitable for improving the nisin production by Lactococcus lactis subsp. lactis W28 was established. The effects of initial sucrose concentration (ISC) in the fermentation broth, feeding time, and feeding rate on the fermentation were studied. It was observed that a fed-batch culture (ISC = 10 g l⁻¹) with 100 ml sucrose solution (190 g l⁻¹) being evenly fed (9–10 ml h⁻¹) into the fermenter after 3-h fermentation gave the best performance in terms of biomass and nisin yield. Under these conditions, the total biomass and the total nisin yield were approximately 23% and 51% higher than those in batch fermentation, respectively. When the sucrose concentration was controlled at 5–10 g l⁻¹ in variable volume intermittent fed-batch fermentation (VVIF) with ISC = 10 g l⁻¹, the total biomass and the total nisin yield were 29% and 60% above those in batch fermentation, respectively. The VVIF proved to be effective to eliminate the substrate inhibition by maintaining sucrose at appropriate levels. It is also easy to be scaled up, since various parameters involved in industrial production were taken into account.     

Pilot-Scale Fermentation of Aqueous-Ammonia-Soaked Switchgrass

Aqueous-ammonia-steeped switchgrass was subject to simultaneous saccharification and fermentation (SSF) in two pilot-scale bioreactors (50- and 350-L working volume). Switchgrass was pretreated by soaking in ammonium hydroxide (30%) with solid to liquid ratio of 5 L ammonium hydroxide per kilogram dry switchgrass for 5 days in 75-L steeping vessels without agitation at ambient temperatures (15 to 33 °C). SSF of the pretreated biomass was carried out using Saccharomyces cerevisiae (D₅A) at approximately 2% glucan and 77 filter paper units per gram cellulose enzyme loading (Spezyme CP). The 50-L fermentation was carried out aseptically, whereas the 350-L fermentation was semiaseptic. The percentage of maximum theoretical ethanol yields achieved was 73% in the 50-L reactor and 52–74% in the 350-L reactor due to the difference in asepsis. The 350-L fermentation was contaminated by acid-producing bacteria (lactic and acetic acid concentrations approaching 10 g/L), and this resulted in lower ethanol production. Despite this problem, the pilot-scale SSF of aqueous-ammonia-pretreated switchgrass has shown promising results similar to laboratory-scale experiments. This work demonstrates challenges in pilot-scale fermentations with material handling, aseptic conditions, and bacterial contamination for cellulosic fermentations to biofuels.     

Effects of Feed Time,Organic Loading and Shock Loads in Anaerobic Whey Treatment by an AnSBBR with Circulation

The aim of this work was to investigate the effect of different feeding times (2, 4, and 6 h) and organic loading rates (3, 6 and 12 gCOD l⁻¹ day⁻¹) on the performance of an anaerobic sequencing batch reactor containing immobilized biomass, as well as to verify the minimum amount of alkalinity that can be added to the influent. The reactor, in which mixing was achieved by recirculation of the liquid phase, was maintained at 30 ± 1°C, possessed 2.5 l reactional volume and treated 1.5 l cheese whey in 8-h cycles. Results showed that the effect of feeding time on reactor performance was more pronounced at higher values of organic loading rates (OLR). During operation at an OLR of 3 gCOD l⁻¹ day⁻¹, change in feeding time did not affect efficiency of organic matter removal from the reactor. At an OLR of 6 gCOD l⁻¹ day⁻¹, reactor efficiency improved in relation to the lower loading rate and tended to drop at longer feeding times. At an OLR of 12 gCOD l⁻¹ day⁻¹ the reactor showed to depend more on feeding time; higher feeding times resulted in a decrease in reactor efficiency. Under all conditions shock loads of 24 gCOD l⁻¹ day⁻¹ caused an increase in acids concentration in the effluent. However, despite this increase, the reactor regained stability readily and alkalinity supplied to the influent showed to be sufficient to maintain pH close to neutral during operation. Regardless of applied OLR, operation with feeding time of 2 h was which provided improved stability and rendered the process less susceptible to shock loads.     

Cultivation of <Emphasis Type="Italic">Rhodobacter sphaeroides</Emphasis> in the Stirred Bioreactor with Different Feeding Strategies for CoQ<Subscript>10</Subscript> Production

The logistic growth model combined with the Luedeking-Piret equation was adopted in this study to model the batch production of CoQ₁₀ in the cultivation of Rhodobacter sphaeroides. The simulation results indicated that CoQ₁₀ production was a primary metabolite. As being a primary metabolite, a longer cell growing stage would tend to accumulate more biomass and lead to a higher CoQ₁₀ concentration being produced. In this context, a fed-batch operation by molasses feeding was performed to increase the biomass and subsequent CoQ₁₀ production. Three different molasses feeding strategies were operated in this study. Results suggested that the fed-batch operation with molasses controlled at 10 ± 1 g/l could increase the cell mass and CoQ₁₀ concentration to reach their maximum values of 18.6 g/l and 83.8 mg/l, respectively, nearly 2.2 times and 1.9 times their respective values obtained in the batch cultivation.     

Biodiesel Residual Glycerol Metabolism by Klebsiella pneumoniae: Pool of Metabolites Under Anaerobiosis and Oxygen Limitation as a Function of Feeding Rates

The metabolism of residual glycerol from biodiesel synthesis by Klebsiella pneumoniae BLh-1 was investigated in this study. Batch and fed-batch cultivations were performed in bioreactors under anaerobic and oxygen limitation conditions. Results of batch cultivations showed that the main product was 1,3-propanediol (1,3-PD) in both conditions, although the higher yields and productivities (0.46 mol mol^?1 glycerol and 1.22 g?L^?1?h^?1, respectively) were obtained under anaerobic condition. Large amounts of ethanol were also produced under batch anaerobic condition, peaking at 12.30 g?L^?1. Batch cultivations under oxygen limitation were characterized by faster growth kinetics, with higher biomass production but lower conversions of glycerol into 1,3-PD, with yields and productivities of 0.33 mol mol^?1 glycerol and 0.99 g?L^?1?h^?1, respectively. The fed-batch cultivations were carried out in order to investigate the effects of feeding of raw glycerol on cells. Fed-batch under anaerobiosis showed that 1,3-PD and ethanol concentrations increased with the feeding rate, with maximal productions of 26.12 and 19.2 g?L^?1, respectively. The oxygen limitation conditions diverted the bacterium metabolism to an elevated lactic acid formation, reaching 59 g?L^?1 in higher feeding rates of glycerol, but lowering the production of ethanol.     

<Emphasis Type="Italic">On-line</Emphasis> Characterization of Metabolic State in Batch Cultivation of <Emphasis Type="Italic">Clostridium diolis</Emphasis> for 1,3-Propanediol Production Using NADH+H<Superscript>+</Superscript> Fluorescence

NADH is a coenzyme which plays a central role in cellular growth and metabolism. It is an intracellular fluorophore which fluoresces at 460 nm when cells are irradiated by 340 nm wavelength of light. The application of NADH+H⁺ fluorescence measurement for characterization of biomass and its metabolic activity during batch fermentation of 1,3-propanediol (1,3-PD) using Clostridium diolis was investigated in this study. A linear correlation between net fluorescence and biomass concentration was observed during both the initial and final phases of 1,3-PD fermentation. This could be used as an on-line indicator of biomass concentration inside the bioreactor thereby eliminating the need for sampling and off-line analysis for establishing biomass concentration during these phases. Also a sharp decline in the NADH+H⁺ fluorescence value was obtained towards the end of fermentation which could be a significant on-line, in situ signal of substrate depletion in the bioreactor and therefore possible fresh nutrient feed for enhanced production of 1,3-PD by repetitive and/or various fed-batch cultivation(s). This is the first report on the use of NADH + H⁺ fluorescence measurement technique for 1,3-PD fermentation.