Versatility of fluorene metabolite (phenol) in fluorene biodegradation by a sulfate reducing culture

Biodegradability of fluorene and the versatility of fluorene metabolite (i.e. phenol) in fluorene biodegradation by a sulfate-reducing enrichment culture were investigated. Batch experiments (with 5 mg l⁻¹ fluorene) were designed via the central composite design to examine the effects of sulfate (5-35 mM) and biomass (5-50 mg l⁻¹) concentrations (variables) on fluorene degradation (response). The experimental results revealed that fluorene removal was more influenced by the biomass concentration than the sulfate concentration. The optimal sulfate and biomass concentrations for fluorene biodegradation (90% removal) were found to be 14.4 mM and 37.8 mg l⁻¹, respectively. Under the optimal conditions, a set of biodegradation experiments were repeated to evaluate both the biodegradability of fluorene metabolite and the potential effect of phenol accumulation on fluorene degradation. The outcomes indicated a slow phenol degradation rate, i.e. 0.02 mg l⁻¹ d⁻¹. Moreover, a small reduction in the fluorene biodegradation efficiency was observed in the presence and accumulation of phenol. However, this sulfate reducing culture is a valuable resource for the simultaneous degradation of fluorene and phenol.     

Biodegradation of phenol using Corynebacterium sp. DJ1 aerobic granules

The single-culture Corynebacterium sp. DJ1 aerobic granules were cultivated and were utilized to degrade high-strength phenolic wastewater. These granules can degrade phenol at sufficient high rate without severe inhibitory effects up to phenol concentration of 2000 mg l⁻¹. Furthermore, the kinetic characteristic noted for these granules yields a zero-order phenol degradation behavior with 500–1500 mg l⁻¹ phenol, which facilitates reactor design and scale up. With added acetate to promote cell growth, this single-culture aerobic granular system yields the highest phenol degradation rate reported in granular literature.     

Phenol removal from hypersaline wastewaters in a Membrane Biological Reactor (MBR): operation and microbiological characterisation

In this study, two Membrane Biological Reactors (MBR) with submerged flat membranes, one at lab-scale conditions and the other at pilot-plant conditions, were operated at environmental temperature to treat an industrial wastewater characterised by low phenol concentrations (8-16 mg L⁻¹) and high salinity (∼150-160 mS cm⁻¹). During the operation of both reactors, the phenol loading rate was progressively increased and less than 1 mg phenol L⁻¹ was detected even at very low HRTs (0.5-0.7 days). Membrane fouling was minimized by the cross flow aeration rate inside the MBRs and by intermittent permeation. Microbial community analysis of both reactors revealed that members of the genera Halomonas and Marinobacter (gammaproteobacteria) were major components. Growth-linked phenol degradation by pure cultures of Marinobacter isolates demonstrated that this bacterium played a major role in the removal of phenol from the bioreactors.     

Phenol degradation performance by isolated Bacillus cereus immobilized in alginate

Phenol degradation by Bacillus cereus AKG1 MTCC9817 and AKG2 MTCC 9818 was investigated and degradation kinetics are reported for the free and Ca-alginate gel-immobilized systems. The optimal pH for maximum phenol degradation by immobilized AKG1 and AKG2 was found to be 6.7 and 6.9, respectively, while 3% alginate was optimum for both the strains. The degradation of phenol by free as well as immobilized cells was comparable at lower concentrations of phenol (100–1000 mg l⁻¹). However, the degradation efficiency of the immobilized strains was higher than that of the free strains at higher phenol concentrations (1500–2000 mg l⁻¹), indicating the improved tolerance of the immobilized cells toward phenol toxicity. More than 50% of 2000 mg l⁻¹ phenol was degraded by immobilized AKG1 and AKG2 within 26 and 36 days, respectively. Degradation kinetics of phenol by free and immobilized cells are well represented by the Haldane and Yano model.     

Biodegradation of phenanthrene and pyrene by Ganoderma lucidum

Biodegradation of two polycyclic aromatic hydrocarbons (PAHs), phenanthrene and pyrene, by a white rot fungus, Ganoderma lucidum, in broth cultures was investigated. It was found that the biomass of the organism decreased with the increase of PAH concentration in the cultures. In the cultures with 2 to 50 mg l⁻¹ PAHs, the degradation rate constants (k₁) increased with the PAH concentration, whereas, at the level of 100 mg l⁻¹, the degradation rate constants decreased. In the presence of 20 mg l⁻¹ PAHs, the highest degradation rates of both PAHs occurred in cultures with an initial pH of 4.0 at 30 °C. The addition of CuSO₄, citric acid, gallic acid, tartaric acid, veratryl alcohol, guaiacol, 2,2′-azino-bis-(3- ethylbenzothazoline-6-sulfonate) (ABTS) enhanced the degradation of both PAHs and laccase activities; whereas the supplement of oxalate, di-n-butyl phthalate (DBP), and nonylphenol (NP) decreased the degradation of both PAHs and inhibited laccase production. In conclusion, G. lucidum is a promising white rot fungus to degrade PAHs such as phenanthrene and pyrene in the environment.     

Anaerobic degradation of tetrachlorobisphenol-A in river sediment

This study investigated the anaerobic degradation of tetrachlorobisphenol-A (TCBPA) in sediment samples collected at three sites along the Erren River in southern Taiwan. TCBPA anaerobic degradation half-lives (t_1/2) in the sediment were 12.6, 16.9 and 21.7 d at concentrations of 50, 100, and 250 ??g g⁻¹, respectively. TCBPA (50 ??g g⁻¹) anaerobic degradation half-lives (t_1/2) in the sediment were 10.1, 11.8, 11.0, 11.6, 10.8, 9.1, 8.5, 18.2, 19.3, and 16.1 d by the addition of yeast extract (5 mg l⁻¹), cellulose (0.96 mg l⁻¹), sodium chloride (1%), brij 30 (130 mg l⁻¹), brij 35 (43 mg l⁻¹), rhamnolipid (55 ??M), surfactin (91 ??M), phthalic esters (2 mg l⁻¹), nonylphenol (2 mg l⁻¹), and heavy metals (2 mg l⁻¹), respectively. The degradation rate of TCBPA was enhanced by the addition of yeast extract, cellulose, sodium chloride, brij 30, brij 35, rhamnolipid, or surfactin. However, it was inhibited by the addition of phthalic esters, nonylphenol, or heavy metals. Also noted was the presence of dichlorobisphenol-A and bisphenol-A, two intermediate products resulting from the anaerobic degradation of TCBPA accumulated in the sediments.     

Degradation kinetics of phenol by immobilized cells of Candida tropicalis in a fluidized bed reactor

Degradation kinetics of phenol by free and agar-entrapped cells of Candida tropicalis was studied in batch cultures. The initial phenol degradation rate achieved with free cells was higher than that obtained with immobilized cells, when phenol concentrations up to 1000 mg l^–1 were used. However, at higher phenol concentrations, the behaviour was quite different. The initial degradation rate of the immobilized yeast cells was about 10 times higher than that of the free cells, at a phenol concentration of 3500 mg l^–1. The semicontinuous and continuous degradation of phenol by immobilized yeast cells was also investigated in a multi-stage fluidized bed reactor. The highest phenol removal efficiencies and degradation rates as well as the lowest values of residual phenol and chemical oxygen demand were obtained in the semicontinuous culture when phenol concentrations up to 1560 mg l^–1 were used.     

Comparison of polysialic acid production in Escherichia coli K1 during batch cultivation and fed-batch cultivation applying two different control strategies

The polysialic acid (PSA) production in Escherichia coli (E. coli) K1 was studied using three different cultivation strategies. A batch cultivation, a fed-batch cultivation at a constant specific growth rate of 0.25 h⁻¹ and a fed-batch cultivation at a constant glucose concentration of 50 mg l⁻¹ was performed. PSA formation kinetics under different cultivation strategies were analyzed based on the Monod growth model and the Luedeking-Piret equation. The results revealed that PSA formation in E. coli K1 was completely growth associated, the highest specific PSA formation rate (0.0489 g g⁻¹ h⁻¹) was obtained in the batch cultivation. However, comparing biomass and PSA yields on the glucose consumed, both fed-batch cultivations provided higher yields than that of the batch cultivation and acetate formation was prevented. Moreover, PSA yield on glucose was also correlated to the specific growth rate of the cells. The optimal specific growth rate for PSA production was 0.32 h⁻¹ obtained in the fed-batch cultivation at a constant glucose concentration of 50 mg l⁻¹, with highest conversion efficiency of 43 mg g⁻¹.     

Benzene, toluene, and o-xylene degradation by free and immobilized P. putida F1 of postconsumer agave-fiber/polymer foamed composites

Benzene, toluene, and o-xylene (BTX) degradation by immobilized Pseudomonas putida F1 of postconsumer agave-fiber/polymer foamed-composites (AFPFC) and suspended cultures was studied under controlled conditions. Analyses using FTIR-ATR and SEM showed that P. putida F1 adhered onto the composite surface and developed a biofilm. In this sense, the AFPFC were successfully used as a support for bacterial immobilization. Both systems, immobilized and suspended cells of P. putida F1, were able to completely degrade benzene and toluene from initial concentrations of 15, 30, 60, and 90 mg l⁻¹. An inhibitory effect of the intermediary catechol from benzene degradation was observed in suspended cultures but it was not presented in the immobilized system. The degradation of o-xylene was partially accomplished in both systems. The Monod equation was used to model the experimental data obtained from the biodegradation kinetics, and they were adequately described with this model.     

Pentachlorophenol (PCP) dechlorination in horizontal-flow anaerobic immobilized biomass (HAIB) reactors

This study verifies the potential applicability of horizontal-flow anaerobic immobilized biomass (HAIB) reactors to pentachlorophenol (PCP) dechlorination. Two bench-scale HAIB reactors (R1 and R2) were filled with cubic polyurethane foam matrices containing immobilized anaerobic sludge. The reactors were then continuously fed with synthetic wastewater consisting of PCP, glucose, acetic acid, and formic acid as co-substrates for PCP anaerobic degradation. Before being immobilized in polyurethane foam matrices, the biomass was exposed to wastewater containing PCP in reactors fed at a semi-continuous rate of 2.0 μg PCP g⁻¹ VS. The applied PCP loading rate was increased from 0.05 to 2.59 mg PCP l⁻¹ day⁻¹ for R1, and from 0.06 to 4.15 mg PCP l⁻¹ day⁻¹ for R2. The organic loading rates (OLR) were 1.1 and 1.7 kg COD m⁻³ day⁻¹ at hydraulic retention times (HRT) of 24 h for R1 and 18 h for R2. Under such conditions, chemical oxygen demand (COD) removal efficiencies of up to 98% were achieved in the HAIB reactors. Both reactors exhibited the ability to remove 97% of the loaded PCP. Dichlorophenol (DCP) was the primary chlorophenol detected in the effluent. The adsorption of PCP and metabolites formed during PCP degradation in the packed bed was negligible for PCP removal efficiency.     

Efficiencies and costs of larval growth in different food environments (Asteroidea: Asterina miniata)

The ocean is a nutritionally heterogeneous environment. For feeding larval forms, food variability has significant consequences for growth and later recruitment success. In this study, the physiological and biochemical responses to a range of different food concentrations (unfed, 4, 20, and 40 algal cells μl^− 1) were examined in larvae of the asteroid, Asterina miniata. Measurements of growth, protein synthesis rates, and the energetic cost of protein synthesis were made. Under conditions of rapid growth, protein comprised a larger percent (66%) of a larva's organic biomass compared to similar-aged, slower-growing larvae (26%). Larvae fed at the highest food concentration tested (40 algal cells μl^− 1) had a protein depositional efficiency of 80% (± 16%), a value 3-fold higher than larvae fed 20 algal cells μl^− 1 (28% ± 11%). Also, faster-growing larvae required 3-fold less energy per unit mass of protein growth. Larvae fed 40 algal cells μl^− 1 deposited protein at a respiratory cost of 65 ± 11 pmol O₂ h^− 1 (μg protein)^− 1; larvae fed 20 algal cells μl^− 1 had a cost of 192 ± 47 pmol O₂ h^− 1 (μg protein)^− 1. While there were differences in the cost to deposit protein (i.e., protein growth, the balance of synthesis and degradation), there were no differences in the energetic cost of protein synthesis for all food concentrations tested. The energetic cost of protein synthesis was fixed at 13.8 (± 0.92) Joules (mg protein synthesized)^− 1 and was independent of developmental stage, growth rates, and large changes (58-fold) in protein synthesis rates. A major conclusion from this study is that larvae grown in high-food environments not only grew faster, but did so for considerably less energy. Defining the complex relationships of food availability and metabolic efficiency will provide more accurate predictions of larval growth under variable food conditions in the ocean.     

Evaluation of a lab-scale tidal flow constructed wetland performance: Oxygen transfer capacity, organic matter and ammonium removal

Oxygen transfer capacity and removal of ammonium and organic matter were investigated in this study to evaluate the performance of a lab-scale tidal flow constructed wetland. Average oxygen supply under tidal operation (350 g m⁻² d⁻¹) was much higher than in conventional constructed wetlands (<100 g m⁻² d⁻¹), resulting in enhanced removal of BOD₅ and NH₄⁺. Theoretical oxygen demand from BOD₅ removal and nitrification was approximately matched by the measured oxygen supply, which indicated aerobic consumption of BOD₅ and NH₄⁺ under tidal operation. When BOD₅ removal increased from 148 g m⁻² d⁻¹ to 294 g m⁻² d⁻¹, neither exhausted oxygen from the aggregate matrix during feeding period (111 g m⁻² d⁻¹) nor effluent dissolved oxygen (DO) concentration (2.8 mg/L) changed significantly, demonstrating that the oxygen transfer potential of the treatment system had not been exceeded. However, even though DO had not been exhausted, inhibition of nitrification was observed under high BOD loading. The loss of nitrification was attributed to excessive heterotrophic biofilm growth believed to induce oxygen transfer limitations or oxygen competition in thickened biofilms.     

Ethanol production from sweet sorghum juice using very high gravity technology: Effects of carbon and nitrogen supplementations

Ethanol production from sweet sorghum juice by Saccharomyces cerevisiae NP01 was investigated under very high gravity (VHG) fermentation and various carbon adjuncts and nitrogen sources. When sucrose was used as an adjunct, the sweet sorghum juice containing total sugar of 280 g l⁻¹, 3 g yeast extract l⁻¹ and 5 g peptone l⁻¹ gave the maximum ethanol production efficiency with concentration, productivity and yield of 120.68 ± 0.54 g l⁻¹, 2.01 ± 0.01 g l⁻¹ h⁻¹ and 0.51 ± 0.00 g g⁻¹, respectively. When sugarcane molasses was used as an adjunct, the juice under the same conditions gave the maximum ethanol concentration, productivity and yield with the values of 109.34 ± 0.78 g l⁻¹, 1.52 ± 0.01 g l⁻¹ h⁻¹ and 0.45 ± 0.01 g g⁻¹, respectively. In addition, ammonium sulphate was not suitable for use as a nitrogen supplement in the sweet sorghum juice for ethanol production since it caused the reduction in ethanol concentration and yield for approximately 14% when compared to those of the unsupplemented juices.     

Multi-objective optimization of glycopeptide antibiotic production in batch and fed batch processes

Fermentation optimization involves potentially conflicting multiple objectives such as product concentration and production media cost. Simultaneous optimization of these objectives would result in a multiobjective optimization problem, which is characterized by a set of multiple solutions, knows as pareto optimal solutions. These solutions gives flexibility in evaluating the trade-offs and selecting the most suitable operating policy. Here, ε-constraint approach was used to generate the pareto solutions for two objectives: product concentration and product per unit cost of media, for batch and fed batch operations using process model for Amycolatopsis balhimycina, a glycopeptide antibiotic producer. This resulted in a set of several pareto optimal solutions with the two objectives ranging from (0.75 g l⁻¹, 3.97 g $^-1) to (0.44 g l⁻¹, 5.19 g $^-1) for batch and from (1.5 g l⁻¹, 5.46 g $^-1) to (1.1 g l⁻¹, 6.34 g $^-1) for fed batch operations. One pareto solution each for batch and for fed batch mode was experimentally validated.     

Assessment of olive-mill wastewater as a growth medium for lipase production by Candida cylindracea in bench-top reactor

Olive-mill wastewater (OMW) was investigated for its suitability to serve as a medium for lipase production by Candida cylindracea NRRL Y-17506. The OMW that best supported enzyme production was characterized by low COD and low total sugars content. In shake flask batch cultures, OMW supplementation with 2.4 g l⁻¹ NH₄Cl and 3 g l⁻¹ olive oil led to an enzyme activity of about 10 U ml⁻¹. The addition of glucose or malt extract and supplements containing organic N (e.g., peptone, yeast extract) either depressed or did not affect the enzyme production. Further experiments were then performed in a 3-l stirred tank reactor to assess the impact of medium pH and stirring speed on the yeast enzyme activity. The lipase activity was low (1.8 U ml⁻¹) when the pH was held constant at 6.5, significantly increased (18.7 U ml⁻¹) with uncontrolled pH and was maximum (20.4 U ml⁻¹) when the pH was let free to vary below 6.5. A stirring regime, that varied depending on the dissolved oxygen concentration in the medium, both prevented the occurrence of anoxic conditions during the exponential growth phase and enabled good lipase production (i.e., 21.6 U ml⁻¹) and mean volumetric productivity (i.e., 123.5 U l⁻¹ h⁻¹).     

Denitrifying sulfide removal and carbon methanogenesis in a mesophilic, methanogenic culture

The inhibitory effects of 90-189 mg l⁻¹ of sulfide and 25-75 mg-N l⁻¹ of nitrate on methanogenesis were investigated in a mixed methanogenic culture using butyrate as carbon source. In the initial phase of 90 mg l⁻¹ S²⁻ test, autotrophic denitrification of nitrate occurred with sulfide as the electron donor. Then the sulfate-reducing strains converted the produced sulfur back to sulfide via heterotrophic oxidation pathway. Methanogenesis was not markedly inhibited when 90 mg l⁻¹ of sulfide was dosed alone. When 25-75 mg-N l⁻¹ of nitrate was presented, initiation of methanogenesis was seriously delayed. Nitrogen oxides (NO_x), the intermediates for nitrate reduction via denitrification pathway, inhibited methanogenesis. The 90 mg l⁻¹ of sulfide favored heterotrophic dissimilatory nitrate reduction to ammonia (DNRA) pathway for nitrate reduction. Possible ways of maximizing methane production from an organic carbon-rich wastewater with high levels of sulfide and nitrate were discussed.     

Sorption of phosphate onto giant reed based adsorbent: FTIR, Raman spectrum analysis and dynamic sorption/desorption properties in filter bed

A sorption process for the removal of phosphate was evaluated under various conditions using a filter bed packed with giant reed (GR) based adsorbent. FTIR spectrum measurement validated the existence of grafted amine groups in the adsorbent and Raman spectrum displayed the characteristic peaks of different forms of phosphate. The column sorption capacity of the adsorbent for phosphate was 54.67 mg g⁻¹ in comparison with the raw GR of 0.863 mg g⁻¹. Influent pH demonstrated an essential effect on the performance of the filter bed as compared to other influent conditions (flow rates and influent concentrations) and the optimal pH was selected at 5.0-10.0. Eluents of HCl, NaOH and NaCl solutions with concentrations of 0.01-0.1 mol l⁻¹ showed the excellent capacities for desorption of phosphate from the adsorbent, and their elution processes could be finished in 90 min.     

Biodegradation kinetics of 2,4,6-Trichlorophenol by an acclimated mixed microbial culture under aerobic conditions

The objective of this study was to achieve a better quantitative understanding of the kinetics of 2,4,6-trichlorophenol (TCP) biodegradation by an acclimated mixed microbial culture. An aerobic mixed microbial culture, obtained from the aeration basin of the wastewater treatment plant, was acclimated in shake flasks utilizing various combinations of 2,4,6-TCP (25–100 mg l⁻¹), phenol (300 mg l⁻¹) and glycerol (2.5 mg l⁻¹) as substrates. Complete primary TCP degradation and a corresponding stoichiometric release of chloride ion were observed by HPLC and IEC analytical techniques, respectively. The acclimated cultures were then used as an inoculum for bench scale experiments in a 4 l stirred-tank reactor (STR) with 2,4,6-TCP as the sole carbon/energy (C/E) source. The phenol acclimated mixed microbial culture consisted of primarily Gram positive and negative rods and was capable of degrading 2,4,6-TCP completely. None of the predicted intermediate compounds were detected by gas chromatography in the cell cytoplasm or supernatant. Based on the disappearance of 2,4,6-TCP, degradation was well modelled by zero-order kinetics which was also consistent with the observed oxygen consumption. Biodegradation rates were compared for four operating conditions including two different initial 2,4,6-TCP concentrations and two different initial biomass concentrations. While the specific rate constant was not dependent on the initial 2,4,6-TCP concentration, it did depend on the initial biomass concentration (X _init). A lower biomass concentration gave a much higher zero-order specific degradation rate. This behaviour was attributed to a lower average biomass age or cell retention time (θ_x) for these cultures. The implications of this investigation are important for determining and predicting the potential risks associated with TCP, its degradation in the natural environment or the engineering implications for ex situ treatment of contaminated ground water or soil.     

Anaerobic degradation of phenol in batch and continuous cultures by a denitrifying bacterial consortium

Summary The anaerobic degradation of phenol under denitrifying conditions by a bacterial consortium was studied both in batch and continuous cultures. Anaerobic degradation was dependent on NO_f3 ^p– and concentrations up to 4 mm phenol were degraded within 2–5 days. During continuous growth in a fermenter, steady states could be maintained at eight dilution rates (D) corresponding to residence times between 12.5 and 50 h. Culture wash-out occurred at D=0.084 h^–1. The kinetic parameters obtained for anaerobic degradation of phenol under denitrifying conditions by the consortium were: maximam specific growth rate = 0.091 h^–1; saturation constant = 4.91 mg phenol/l; true growth yield = 0.57 mg dry wt/mg phenol; maintenance coefficient = 0.013 mg phenol/mg dry wt per hour. The Haldane model inhibition constant was estimated from batch culture data giving a value of 101 mg/l. The requirement of CO₂ for the anaerobic degradation of phenol with NO_f3 ^p– indicates that phenol carboxylation to 4-hydroxybenzoate was the first step of phenol degradation by this culture. 4-Hydroxybenzoate, proposed as an intermediate of phenol carboxylation under these conditions, was detected only in continuous cultures at very low growth rates (D=0.02 h^–1), but was never detected as a free intermediary metabolite either in batch or in continuous cultures. Correspondence to: N. Khoury     

Synergistic production of L-arabinose from arabinan by the combined use of thermostable endo- and exo-arabinanases from Caldicellulosiruptor saccharolyticus

The optimum conditions for the production of l-arabinose from debranched arabinan were determined to be pH 6.5, 75 °C, 20 g l⁻¹ debranched arabinan, 42 U ml⁻¹ endo-1,5-α-l-arabinanase, and 14 U ml⁻¹ α-l-arabinofuranosidase from Caldicellulosiruptor saccharolyticus and the conditions for sugar beet arabinan were pH 6.0, 75 °C, 20 g l⁻¹ sugar beet arabinan, 3 U ml⁻¹ endo-1,5-α-l-arabinanase, and 24 U ml⁻¹ α-l-arabinofuranosidase. Under the optimum conditions, 16 g l⁻¹l-arabinose was obtained from 20 g l⁻¹ debranched arabinan or sugar beet arabinan after 120 min, with a hydrolysis yield of 80% and a productivity of 8 g l⁻¹ h⁻¹. This is the first reported trial for the production of l-arabinose from the hemicellulose arabinan by the combined use of endo- and exo-arabinanases.