Biodegradation kinetics of peptone and 2,6-dihydroxybenzoic acid by acclimated dual microbial culture

This study evaluated the kinetics of simultaneous biodegradation of peptone mixture and 2,6-dihydroxybenzoic acid (2,6-DHBA) by an acclimated dual microbial culture under aerobic conditions. A laboratory-scale sequencing batch reactor was sustained at steady-state with peptone mixture feeding. During the study period, peptone mixture feeding was continuously supplemented with 2,6-DHBA. Related experimental data were derived from three sets of parallel batch reactors, the first fed with the peptone mixture, the second with 2,6-DHBA and the third one with the two substrates, after acclimation of microbial culture and simultaneous biodegradation of both organics. A mechanistic model was developed for this purpose including the necessary model components and process kinetics for the model calibration of relevant experimental data. Model evaluation provided all biodegradation characteristics and kinetics for both peptone mixture and 2,6-DHBA. It also supported the development of a dual microbial community through acclimation, with the selective growth of a second group of microorganisms specifically capable of metabolizing 2,6-DHBA as an organic carbon source.     

Bioremediation 2,4,6,-Trichlorophenol (2,4,6-TCP) by Shigella sp. S2 Isolated from Industrial Dumpsite

A bacterium that utilizes 2,4,6-trichlorophenol (2,4,6-TCP) as a sole source of carbon and energy was isolated from an industrial dumpsite, the bacterium designated as strain S2. Degradation was routinely monitored by observing growth analysis, chloride release assay, and ring cleavage activity and was further confirmed by gas chromatography (GC) analysis. The bacterium was found to degrade up to 90% of 2,4,6-TCP at 1.5 mM concentration. The bacteria were characterized morphologically, biochemically, and by 16S rRNA gene sequencing, which showed 99% sequence similarity with Shigella sp. This is the first report that Shigella sp. was able to degrade 2,4,6-TCP. This strain was found to be novel and a potential 2,4,6-TCP degrader. Further, this strain may be used for bioremediation of 2,4,6-TCP–containing waste in the environment.     

Biodegradation kinetics of methyl iso-butyl ketone by acclimated mixed culture

Methyl iso-butyl ketone (MIBK) is a widely used volatile organic compound (VOC) which is highly toxic in nature and has significant adverse effects on human beings. The present study deals with the removal of MIBK using biodegradation by an acclimated mixed culture developed from activated sludge. The biodegradation of MIBK is studied for an initial MIBK concentration ranging from 200–700 mg l⁻¹ in a batch mode of operation. The maximum specific growth rate achieved is 0.128 h⁻¹ at 600 mg l⁻¹of initial MIBK concentration. The kinetic parameters are estimated using five growth kinetic models for biodegradation of organic compounds available in the literature. The experimental data found to fit well with the Luong model (R ² = 0.904) as compared to Haldane model (R ² = 0.702) and Edward model (R ² = 0.786). The coefficient of determination (R ²) obtained for the other two models, Monod and Powell models are 0.497 and 0.533, respectively. The biodegradation rate found to follow the three-half-order kinetics and the resulting kinetic parameters are reported.     

Polyhydroxybutyrate production from lactate using a mixed microbial culture

In this study we investigated the use of lactate and a lactate/acetate mixture for enrichment of poly-3-hydroxybutyrate (PHB) producing mixed cultures. The mixed cultures were enriched in sequencing batch reactors (SBR) that established a feast-famine regime. The SBRs were operated under conditions that were previously shown to enable enrichment of a superior PHB producing strain on acetate (i.e., 12 h cycle length, 1 day SRT and 30°C). Two new mixed cultures were eventually enriched from activated sludge. The mixed culture enriched on lactate was dominated by a novel gammaproteobacterium. This enrichment can accumulate over 90 wt% PHB within 6 h, which is currently the best result reported for a bacterial culture in terms of the final PHB content and the biomass specific PHB production rate. The second mixed culture enriched on a mixture of acetate and lactate can produce up to 84 wt% PHB in just over 8 h. The predominant bacterial species in this culture were Plasticicumulans acidivorans and Thauera selenatis, which have both been reported to accumulate large amounts of PHB. The data suggest that P. acidivorans is a specialist on acetate conversion, whereas Thauera sp. is a specialist on lactate conversion. The main conclusion of this work is that the use of different substrates has a direct impact on microbial composition, but has no significant effect on the functionality of PHB production process.     

Biodegradation of polyvinyl alcohol by a mixed microbial culture

A mixed culture capable of degrading 1 g l⁻¹ polyvinyl alcohol (PVA) completely was screened from sludge samples at Pacific Textile Factory, Wuxi, China. This mixed culture had stronger capability of degrading PVA with low polymerization and high saponification than degrading PVA with high polymerization and low saponification. Inorganic nitrogen source was more suitable for the mixed culture to grow and degrade PVA than organic nitrogen source. Microorganisms and relative abundance of this mixed culture were explored by terminal restriction fragment length polymorphism (T-RFLP). Small PVA molecules were detected in cell extracts of the mixed culture. This indicated that PVA degradation in the mixed culture was in fact a combined action of extracellular and intracellular enzymes. Two strains producing extracellular PVA-degrading enzyme were isolated from the mixed culture. They could individually degrade PVA1799 with polymerization of 1700 from initial average molecular weight 112,981 to 98,827 Da and 84,803 Da, respectively. However, only small amount of PVA124 in polymerization of 400 could be degraded by these two strains.     

Biodegradation kinetics of stored wastewater substrates by a mixed microbial culture

The anaerobic accumulation of several organic pollutants from industrial wastewaters, as storage substrates, and their subsequent aerobic biodegradation using a wastewater treatment mixed microbial culture for biological nutrient removal has been studied. The amount and the kinetics of substrate accumulation in the anaerobic stage depended on the characteristics of the wastewater fed to the anaerobic stage. Depending on the substrate used, levels of between 27 and 86% of storage polymers were accumulated with respect to the level obtained on feeding with acetate. The biodegradation kinetics were studied by modelling respirometry results. During the aerobic stage, oxygen-consumption data obtained in the respirometric tests were fitted to a model using a non-linear fitting estimation method. The simulation data obtained correlated well with the experimental oxygen-consumption data. The estimated kinetic parameters obtained indicate that each storage polymer was degraded at a different rate. However, the values obtained for the storage polymer half-saturation coefficient, K_S: 16 mg COD l⁻¹, and for the coefficient for endogenous respiration, b: 0.008 h⁻¹, were similar in all the experiments. The results indicate that each substrate produces the synthesis of a specific storage polymer that is degraded at a different rate.     

Biodegradation of 2,4,6-Tribromophenol by Ochrobactrum sp. Strain TB01

A bacterium that utilizes 2,4,6-tribromophenol (2,4,6-TBP) as sole carbon and energy source was isolated from soil contaminated with brominated pollutants. This bacterium, designated strain TB01, was identified as an Ochrobactrum species. The organism degraded 100 μM of 2,4,6-TBP within 36 h in a growing culture. In addition, it released 3 mol of bromine ions from 1 mol of 2,4,6-TBP during the complete degradation of 2,4,6-TBP in a resting cell assay. Moreover, cells grown on 2,4,6-TBP degraded 2,6-dibromophenol (2,6-DBP), 4-bromophenol (4-BP), 2,4,6-trichlorophenol (2,4,6-TCP) and phenol. Metabolic intermediates were detected in the reaction mixture of an in vitro assay for 2,4,6-TBP, and they were identified as 2,4-DBP and 2-BP. NADH was required for the debromination of 2,4,6-TBP. These results suggest that 2,4,6-TBP is converted to phenol through sequential reductive debromination reactions via 2,4-DBP and 2-BP by this strain.     

Biodegradation of polyethylene glycol by symbiotic mixed culture (obligate mutualism)

Neither Flavobacterium sp. nor Pseudomonas sp. grew on a polyethylene glycol (PEG) 6000 medium containing the culture filtrate of their mixed culture on PEG 6000. The two bacteria did not grow with a dialysis culture on a PEG 6000 medium. Flavobacterium sp. grew well on a dialysis culture containing a tetraethylene glycol medium supplemented with a small amount of PEG 6000 as an inducer, while poor growth of Pseudomonas sp. was observed. Three enzymes involved in the metabolism of PEG, PEG dehydrogenase, PEG-aldehyde dehydrogenase and PEG-carboxylate dehydrogenase (ether-cleaving) were present in the cells of Flavobacterium sp. The first two enzymes were not found in the cells of Pseudomonas sp. PEG 6000 was degraded neither by intact cells of Flavobacterium sp. nor by those of Pseudomonas sp., but it was degraded by their mixture. Glyoxylate, a metabolite liberated by the ether-cleaving enzyme, inhibited the growth of the mixed culture. The ether-cleaving enzyme was remarkably inhibited by glyoxylate. Glyoxylate was metabolized faster by Pseudomonas sp. than by Flavobacterium sp., and seemed to be a key material for the symbiosis.     

Biodegradation and metabolites of 2-methylquinoline by acclimated activated sludge under aerobic and denitrifying conditions

2-Methylquinoline is a common organic contaminant in environment. Its degradation in wastewater treatment system has not been fully explored. In this study, batch experiments were conducted to investigate the biodegradation of 2-methylquinoline by activated sludge under both aerobic and denitrifying conditions. The results showed that 2-methylquinoline was degraded under both conditions, but the degradation under aerobic condition was significantly faster than that under denitrifying condition. Total organic carbon (TOC) residues were detected in the final effluent under both conditions, indicating the formation of recalcitrant metabolites. Further analysis identified 1,2,3,4-tetrahydro-2-methylquinoline, N,N-diethyl-benzenamine, and 4-ethyl-benzenamine as common metabolites under both conditions. 4-Butyl-benzenamine and 2,6-diethyl-benzenamine were additional metabolites under the aerobic condition, whereas 2-methyl-4-quinolinol was exclusive to the denitrifying condition. Most of these metabolites were further degraded during the treatment process. 1,2,3,4-Tetrahydro-2-methylquinoline, however, remained in the final effluent under both conditions, implying its persistence in the environment. It can be concluded that 2-methylquinoline undergoes the similar degradation pathway under both treatment conditions.     

Biodegradation of 2,4,5-trichlorophenol by aerobic microbial communities: biorecalcitrance,inhibition, and adaptation

Chlorinated aromatic compounds challenge our environment and wastewater treatment processes due to their biorecalcitrance and inhibition. In particular, 2,4,5-trichlorophenol (TCP) seems to demonstrate greater resistance to biodegradation than other trichlorophenols and is a known uncoupler of the electron transport chain, although little work addresses this compound specifically. Here, we investigate the biorecalcitrance, inhibition, and adaptation to 2,4,5-trichlorophenol by aerobic mixed microbial communities. We show that 2,4,5-trichlorophenol is strongly resistant to biodegradation at concentrations greater than 40 μM, demonstrates inhibition to respiration in direct proportion to 2,4,5-trichlorophenol concentration (with 50% inhibition projected near 85 μM 2,4,5-trichlorophenol), and does not sustain biomass in continuous reactors, even when all input 2,4,5-trichlorophenol is degraded. Communities showed consistent adaptation patterns to 2,4,5-trichlorophenol at concentrations of 10 μM and 20 μM, but these patterns diverged at concentrations greater than 40 μM. Finally, thermodynamic approximations were used to estimate the yield of 2,4,5-trichlorophenol as 0.165 gVSS/gCOD, a low value that partially explains why biodegradation of 2,4,5-trichlorophenol did not sustain the biomass. In particular, we estimated that the minimum concentration to support steady-state biomass (S _min) is approximately 180 μM, a value much larger than the 40-μM concentration that is strongly resistant to biodegradation. Thus, readily biodegradable concentrations of 2,4,5-trichlorophenol are too low to sustain the biomass that biodegrades it.     

Degradation of biphenyl by methanogenic microbial consortium

Biphenyl was readily degraded and mineralized to CO₂ and CH₄ by a PCB-dechlorinating anaerobic microbial consortium. Degradation occurred when biphenyl was supplied as a sole source of carbon or as a co-metabolic substrate together with glucose and methanol. p-Cresol was detected and confirmed by mass spectroscopy as a transient intermediate. Production of ¹⁴ C-CO₂ and ¹⁴C-CH₄ from ¹⁴C-biphenyl was observed in the approximate ratio of 1:2. The results indicated the existence of novel pathways for biphenyl degradation in a natural anaerobic microbial community.     

Biodegradation of 4-nitrophenol by indigenous microbial populations in Everglades soils

The Everglades in South Florida are a unique ecologicalsystem. As a result of the widespread use of pesticides andherbicides in agricultural areas upstream from these wetlands,there is a serious potential for pollution problems in theEverglades. The purpose of this study was to evaluate theability of indigenous microbial populations to degradexenobiotic organic compounds introduced by agricultural andother activities. Such biodegradation may facilitate theremediation of contaminated soils and water in the Everglades.The model compound selected in this study is 4-nitrophenol, achemical commonly used in the manufacture of pesticides. Themineralization of 4-nitrophenol at various concentrations wasstudied in soils collected from the Everglades. Atconcentrations of 10 and 100 µg/g soil, considerablemineralization occurred within a week. At a higherconcentration, i.e., 10 mg/g soil, however, no mineralizationof 4-nitrophenol occurred over a 4-month period; such a highconcentration apparently produced an inhibitory effect. Therate and extent of 4-nitrophenol mineralization was enhancedon inoculation with previously isolated nitrophenol-degradingmicroorganisms. The maximum mineralization extent measured,however, was less than 30% suggesting conversion to biomassand/or unidentified intermediate products. These resultsindicate the potential for natural mechanisms to mitigate theadverse effects of xenobiotic pollutants in a complex systemsuch as the Everglades.     

Transformation of 2,4,6-trinitrotoluene (TNT) by immobilized Phanerochaete chrysosporium under fed-batch and continuous TNT feeding conditions

The cometabolic transformation of 2,4,6-trinitrotoluene (TNT) by an immobilized Phanerochaete chrysosporium culture was investigated under different TNT and/or glycerol feeding conditions in a 5-L reactor. In the fed-batch feeding mode, as a result of four spiking events at an average feeding rate of 20 mg TNT L(-1) d(-1) and 250 mg glycerol L(-1) d(-1), the initial TNT transformation rate and the glycerol uptake rate of the 7-day-old immobilized cell culture were 2.41 mg L(-1) h(-1) and 16.6 mg L(-1) h(-1), respectively. Thereafter, the TNT fed into the reactor depicted a negative effect on the cell physiology of P. chrysosporium, i.e., both rates decreased constantly. At 32 mg TNT L(-1) d(-1) feeding rate, also in the presence of glycerol (200 mg L(-1) d(-1)), this effect on the fungal cell metabolism was even more significant. When TNT was fed alone at 3.7 mg L(-1) d(-1), it showed an initial 0.75 mg L(-1) h(-1) rate of TNT transformation, i.e., one-third the initial level observed in the presence of glycerol. In contrast, in the continuous feeding mode (dilution rate, D = 0.11 d(-1)), at 5.5 mg TNT L(-1) d(-1) and 220 mg glycerol L(-1) d(-1), the immobilized cell culture exhibited a constant TNT transformation rate for cultivation periods of 50 and 61 days, under uncontrolled and controlled pH conditions, respectively. Thereafter, during the latter experiment, 100% TNT biotransformation was achieved at 1,100 mg L(-1) d(-1) glycerol feeding rate. Immobilized cells (115-day-old), sampled from a continuous TNT feeding experiment, mineralized [(14)C]-TNT to a level of 15.3% following a 41-day incubation period in a microcosm.     

Degradation of 2,4,6-trichlorophenol (2,4,6-TCP) by co-immobilization of anaerobic and aerobic microbial communities in an upflow reactor under air-limited conditions

The co-immobilization and the culture of anaerobic and aerobic communities was tested for the mineralization of 2,4,6-trichlorophenol (2,4,6-TCP). At first, the anaerobic microorganisms (aggregated into granules) were cultivated in an upflow anaerobic sludge blanket (UASB) reactor, in a continuous mode, with glucose, propionate, acetate (COD loading rate = 0.5-2.0 g COD/l per day, ratio 1:1:1) and 2,4,6-TCP (2,4,6-TCP loading rate = 25-278 micromol/l per day) as substrates. 2,4,6-TCP was degraded into 2,4-DCP and 4-CP, but it was not mineralized because of the low degradation rates of 4-CP. Furthermore, the highest loading rates of 2,4,6-TCP (>126 micromol/l per day) caused the inhibition of the strains degrading the propionate. The granules were therefore tested in association with the aerobic community. They were immobilized in kappa-carrageenan/gelatin [2% (w/w) of each polymer] gel beads and cultivated in a reactor, on their own (to test the influence of the gel), and then with the aerobic community, under anaerobic and air-limited conditions, respectively. The results showed that (1) the gel did not influence the activity of the granules, (2) the anaerobic and aerobic communities could be easily co-immobilized in gel beads and cultivated in a reactor, (3) the mineralization of 2,4,6-TCP (2,4,6-TCP loading rate = 10-506 micromol/l per day), its intermediates of degradation and the other substrates [glucose + acetate + propionate (ratio 1:1:1) = COD loading rate = 500 mg COD/l per day] could be obtained under air-limited conditions if the culture parameters were strictly controlled [airflow = 36-48 vvd (volume of air/volume of liquid in the reactor per day), pH value at around 7.5]. Finally, the gel did not retain its structure during the whole culture (263 days) in the air-limited reactor, but the anaerobic and aerobic communities retained their activities and worked together for the mineralization.     

Biodegradation of p-aminoazobenzene by Bacillus subtilis under aerobic conditions

Biological oxidation of organic dyes is important for textile industry wastewater treatment. The aim of this work was to assess the biodegradation kinetics of a specific azo-dye, p-aminoazobenzene. The degradation of p-aminoazobenzene by Bacillus subtilis was examined through batch experiments in order to investigate the effect of p-aminoazobenzene on the bacterial growth rate and elucidate the mechanism of dye degradation. The results proved that B. subtilis cometabolizes p-aminoazobenzene in the presence of glucose as carbon source, producing aniline and p-phenylenediamine as the nitrogen–nitrogen double bond is broken. The azo-dye was found to act as an inhibitor to microbial growth. A mathematical model was developed that describes cellular growth, glucose utilization, p-aminoazobenzene degradation and product formation. Received 26 July 1996/ Accepted in revised form 14 May 1997     

共代谢条件下光合细菌对2-氯苯酚的生物降解

光合细菌PSB-1D不能利用2-氯苯酚(2-CP)作为唯一的碳源和能源.选用苹果酸、丙酸钠、乙酸钠、柠檬酸钠、苯酚、葡萄糖和可溶性淀粉等7种不同碳源作为光合细菌PSB-1D降解2-CP的共代谢基质,考察了在黑暗好氧培养条件下,不同共代谢基质对PSB-1D生长及降解2-CP效果的影响.结果表明:葡萄糖能够很好地促进PSB-1D的大量繁殖,提高降解效果,缩短降解周期,为最佳共代谢基质.对葡萄糖的投加浓度进行了优化,当葡萄糖的投加浓度为3 g·L-1时,菌株PSB-1D培养168 h后的菌体生长浓度△D560为1.749,2-CP的半衰期为3.9 d,降解速率常数为0.00864 h-1.采用SDS-PAGE对微生物全细胞蛋白质进行分析发现,在共代谢过程中当菌株PSB-1D利用葡萄糖作为底物提供能源和碳源时,可诱导产生2-CP特异性降解酶.     

Biodegradation of aromatic compounds under mixed oxygen/denitrifying conditions: a review

Bioremediation of aromatic hydrocarbons in groundwater and sediments is often limited by dissolved oxygen. Many aromatic hydrocarbons degrade very slowly or not at all under anaerobic conditions. Nitrate is a good alternative electron acceptor to oxygen, and denitrifying bacteria are commonly found in the subsurface and in association with contaminated aquifer materials. Providing both nitrate and microaerophilic levels of oxygen may result in oxidation of the stable benzene rings in aromatic contaminants and allow for the intermediates of this oxidation to degrade via denitrification. The effects of using mixed electron acceptors on biodegradation of subsurface contaminants is unclear. Below some critical oxygen threshold, aerobic biodegradation is inhibited, however high levels of oxygen inhibit denitrification. The mechanisms which regulate electron transfer to oxygen and nitrate are complex. This review: 1) describes the factors which may affect the utilization of oxygen and nitrate as dual electron acceptors during biodegradation; 2) summarizes the incidence of dual use of nitrate and oxygen (aerobic denitrification); and 3) presents evidence of the effectiveness of bioremediation under mixed oxygen/nitrate conditions. Received 08 November 1995/ Accepted in revised form 09 June 1996     

Stoichiometry and kinetics of microbial toluene degradation under denitrifying conditions

Batch experiments were carried out to investigate the stoichiometry and kinetics of microbial degradation of toluene under denitrifying conditions. The inoculum originated from a mixture of sludges from sewage treatment plants with alternating nitrification and denitrification. The culture was able to degrade toluene under anaerobic conditions in the presence of nitrate, nitrite, nitric oxide, or nitrous oxide. No degradation occurred in the absence of Noxides. The culture was also able to use oxygen, but ferric iron could not be used as an electron acceptor. In experiments with¹⁴C-labeled toluene, 34%±8% of the carbon was incorporated into the biomass, while 53%±10% was recovered as¹⁴CO₂, and 6%±2% remained in the medium as nonvolatile water soluble products. The average consumption of nitrate in experiments, where all the reduced nitrate was recovered as nitrite, was 1.3±0.2 mg of nitrate-N per mg of toluene. This nitrate reduction accounted for 70% of the electrons donated during the oxidation of toluene. When nitrate was reduced to nitrogen gas, the consumption was 0.7±0.2 mg per mg of toluene, accounting for 97% of the donated electrons. Since the ammonia concentration decreased during degradation, dissimilatory reduction of nitrate to ammonia was not the reductive process. The degradation of toluene was modelled by classical Monod kinetics. The maximum specific rate of degradation, k, was estimated to be 0.71 mg toluene per mg of protein per hour, and the Monod saturation constant, K _s , to be 0.2 mg toluene/l. The maximum specific growth rate, _max , was estimated to be 0.1 per hour, and the yield coefficient, Y, was 0.14 mg protein per mg toluene.Abbreviations NVWP Non Volatile Water-soluble Products     

Microbial biomass and growth kinetics of microorganisms in chernozem soils under different land use modes

The carbon content of microbial biomass and the kinetic characteristics of microbial respiration response to substrate addition have been estimated for chernozem soils under different land use: arable lands used for 10, 46, and 76 years, mowed meadow, natural forest, and forest shelter belt. Microbial biomass and the content of microbial carbon in humus (C_mic /C_org) decreased in the following order: soils under forest cenoses—mowed meadow—10-year arable land—46- and 75-year arable land. The amount of microbial carbon in the long-plowed horizon was 40% of its content in the upper horizon of natural forest. Arable soils were characterized by a lower metabolic diversity of microbial community and by the highest portion of microorganisms able to grow directly on glucose introduced into soil. The effects of different scenarios of carbon sequestration in soil on the amounts and activity of microbial biomass are discussed.     

Biodegradation of phenanthrene by the indigenous microbial biomass in a zinc amended soil

AIMS: To study the effect of zinc on the biodegradation of phenanthrene by the microbial biomass in soil. METHODS AND RESULTS: Uncontaminated soil was amended with zinc and phenanthrene as single or co-contaminants, and microbial metabolic activity was measured using an intracellular dehydrogenase enzyme bioassay over 37 days. Contaminants were amended at optimum, action and double the action level specified in 'The New Dutch List' (Ministry of Housing, Spatial Planning and Environment, the Netherlands, 2000). Microbial activity in soils with zinc or phenanthrene alone indicated the presence of tolerant, albeit inhibited soil micro-organisms. A zinc concentration at the optimum level of 140 mg kg(-1) in the co-contaminated soil (phenanthrene at 40 mg kg(-1)) resulted in marginal stimulation of the rate of phenanthrene biodegradation. However, Zn2+ concentrations at the action and double the action level of zinc (720 and 1440 mg kg(-1)) inhibited phenanthrene degradation. CONCLUSIONS: Biodegradation of phenanthrene in soils co-contaminated with zinc at concentrations above the action value is impeded. SIGNIFICANCE AND IMPACT OF THE STUDY: Bioremediation efforts to remove polycyclic aromatic hydrocarbon in zinc co-contaminated soils are likely to be constrained.