Enhanced degradation of naphthalene by immobilization of Pseudomonas sp. strain NGK1 in polyurethane foam

A Pseudomonas sp. strain NGKI (NCIM 5120) capable of degrading naphthalene was immobilized in polyurethane foam. The naphthalene-degrading activity of the freely suspended cells was compared with that of immobilized cells in batches in shaken culture and in a continuous culture system in a packed-bed reactor. Increasing concentrations of naphthalene were better tolerated and more quickly degraded by immobilized cell cultures than by free cells. An initial naphthalene concentration of 25 mM was completely degraded by freely suspended cells (4 x 10(10) cfu ml(-1)) and polyurethane-foam-immobilized cells (0.8-1 x 10(12) cfu g(-1) foam cubes) after 4 days and 2 days of incubation, respectively. Free cells degraded a maximum of 30 mM naphthalene after 4 days of incubation with 50 mM naphthalene, and no further degradation was observed even after 15 days of incubation, whereas foam-immobilized cells brought about the complete degradation of 50 mM initial naphthalene after 6 days of incubation. Furthermore, with 25 mM naphthalene, the polyurethane-foam-immobilized cells were re-used 45 times over a period of 90 days without losing naphthalene-degrading activity. By contrast, with the same amount of naphthalene, alginate-, agar-, and polyacrylamide-entrapped cells could be reused for 18, 12, and 23 times over a period of 44, 28, and 50 days, respectively. During continuous degradation in a packed-bed reactor, foam-immobilized cells degraded 80 mM naphthalene at a rate of 150 ml(-1) h(-1). With the same flow rate and 40 mM naphthalene, this system operated efficiently and continuously for about 120 days, whereas the packed-bed reactor with alginate-, agar-, and polyacrylamide-entrapped cells could be operated only for 45, 40, and 60 days respectively. Thus, more efficient degradation of naphthalene could be achieved by immobilizing cells of Pseudomonas sp. strain NGK1 in polyurethane foam, rather than in the other matrices tested.     

Oily sludge degradation by bacteria from Ankleshwar, India

Three bacterial strains, Bacillus sp. SV9, Acinetobacter sp. SV4 and Pseudomonas sp., SV17 from contaminated soil in Ankleshwar, India were tested for their ability to degrade the complex mixture of petroleum hydrocarbons (such as alkanes, aromatics, resins and asphaltenes), sediments, heavy metals and water known as oily sludge. Gravimetric analysis showed that Bacillus sp. SV9 degraded approx. 59% of the oily sludge in 5 days at 30 °C whereas Acinetobacter sp. SV4 and Pseudomonas sp. SV17 degraded 37% and 35%. Capillary gas chromatographic analysis revealed that after 5 days the Bacillus strain was able to degrade oily sludge components of chain length C₁₂–C₃₀ and aromatics more effectively than the other two strains. Maximum drop in surface tension (from 70 to 28.4 mN/m) was accompanied by maximum biosurfactant production (6.7 g l⁻¹) in Bacillus sp. SV9 after 72 h, these results collectively indicating that this bacterial strain has considerable potential for bioremediation of oily sludge.     

Degradation of 2-methylnaphthalene by free and immobilized cells of Pseudomonas sp. strain NGK1

A Pseudomonas sp. strain NGK1 (NCIM 5120) capable of utilizing 2-methylnaphthalene (2-MN) was immobilized in various matrices namely, polyurethane foam (PUF), alginate, agar and polyvinyl alcohol (PVA) (1.5 × 10¹² c.f.u. g^–1 beads). The degradation rates of 25 and 50 mM 2-MN by freely suspended cells (2 × 10¹¹ c.f.u. ml^–1) and immobilized cells in batches, semi-continuous with shaken culture and continuous degradation in a packed-bed reactor were compared. The PUF-immobilized cells achieved higher degradation of 25 and 50 mM of 2-MN than freely suspended cells and the cells immobilized in alginate, agar or PVA. The PVA- and PUF-immobilized cells could be reused for more than 30 and 20 cycles respectively, without losing any degradation capacity. The effect of dilution rates on the rate of degradation of 25 and 50 mM 2-MN with freely suspended and immobilized cells were compared in the continuous system. Increase in dilution rate increased the degradation rate only up to 1 h^–1 in free cells with 25 mM 2-MN and no significant increase was observed with 50 mM 2-MN. With immobilized cells, the degradation rate increased with increase in dilution rate up to 1.5 h^–1 for 25 mM and 1 h^–1 for 50 mM 2-MN. These results revealed that the immobilized cell systems are more efficient than freely suspended cells for biodegradation of 2-MN.     

Enhanced degradation of petrol (Slovene diesel) in an aqueous system by immobilized Pseudomonas fluorescens

The rate of degradation of n -alkanes C₁₂-C₁₈, in petrol (Slovene diesel) in an aqueous system, by free and immobilized Pseudomonas fluorescens in shaking flasks was investigated. Cells were immobilized to a biosupport, Biofix, and a biosorbant, Drizit. Analysis of cellular growth of the free and immobilized bacteria over 8 d of incubation with diesel as the sole carbon source, showed a reduction in the lag phase in the immobilized cultures in comparison to the free system. The free system degraded 52·3% of C₁₂ and 11·6% of C₁₃, but C₁₄-C₁₈ were not degraded. In comparison to the free system and diesel which had not been exposed to experimental conditions (unexposed), the immobilized systems degraded significantly more of C₁₃-C₁₈. Biofix-immobilized cells degraded 14·8% of C₁₂ and an average of 53·5% of C₁₃-C₁₈. Drizit-immobilized cells degraded 24·5% of C₁₂, 52·4% of C₁₃ and an average of 91·2% of C₁₄-C₁₈. This study shows the successful use of immobilized bacteria technology to enhance the degradation of diesel in an aqueous system.     

Improving the biotreatment of hydrocarbons-contaminated soils by addition of activated sludge taken from the wastewater treatment facilities of an oil refinery

Addition of activated sludge taken from the wastewater treatment facilities ofan oil refinery to a soil contaminated with oily sludge stimulated hydrocarbonbiodegradation in microcosms, bioreactors and biopile. Microcosms containing50 g of soil to which 0.07 % (w/w) of activated sludge was added presented ahigher degradation of alkanes (80 % vs 24 %) and polycyclic aromatic hydrocarbons(PAHs) (77 % vs 49 %) as compared to the one receiving only water, after 30days of incubation at room temperature. Addition of ammonium nitrate or sterilesludge filtrate instead of activated sludge resulted in a similar removal of PAHsbut not of alkanes suggesting that the nitrogen contained in the activated sludgeplays a major role in the degradation of PAHs while microorganisms of thesludge are active against alkanes. Addition of sludge also stimulated hydrocarbonbiodegradation in 10-kg bioreactors operated during 60 days and in a 50-m³ biopile operated during 126 days. This biopile treatment allowed the use of the soil for industrial purpose based on provincial regulation (``C' criteria). In contrast, the soil of the control biopile that received only water still exceeded C criteria for C₁₀–C₅₀ hydrocarbons, total PAHs, chrysene and benzo[a]anthracene.The stimulation effect of sludge was stronger on the 4-rings than on 2-rings PAHs.The soil of the biopile that received sludge was 4–5 times less toxic than the control. These results suggest that this particular type of activated sludge could be used to increase the efficiency of the treatment of hydrocarbon-contaminated soils in a biopile.     

Biodegradation of Carbofuran phenol by free and immobilized cells of <Emphasis Type="Italic">Klebsiella pneumoniae</Emphasis> ATCC13883T

The Klebsiella sp. strain ATCC13883T capable of degrading carbofuran phenol (2,3-dihydro-2,2-dimethylbenzofuran-7-ol) has been separated from the soil by enrichment culture technique and immobilized in various, namely polyurethane foam (PUF), polyacrylamide, alginate, agar and alginate-bentonite clay-powdered activated charcoal (PAC). The degradation rates of 20 and 30 mM carbofuran phenol by free and immobilized cells in batch and semi-continuous shaken cultures were compared. The PUF-immobilized cells achieved higher degradation rates in a shorter time than freely suspended cells and the cells immobilized in polyacrylamide, alginate and agar. The PUF- and alginate-bentonite clay-PAC-immobilized cells could be reused for more than 36 cycles, polyacrylamide-entrapped cells for 20 cycles and alginate-bentonite-PAC 28 cycles, without losing any degradation capacity and showed better tolerance to pH, temperature and concentration changes than free cells. These results showed that cells immobilized in modified alginate-bentonite-PAC immobilizers tolerated and completely degraded carbofuran phenol at initial concentrations of 20 and 30 mM and also higher. Such a bacterial strain could be used for bioremediation of environments contaminated with phenolic compounds.     

Biodegradation of Total Petroleum Hydrocarbons from Oily Sludge

The present study concerns the biotreatment of oily sludge of petroleum refineries. Experiments were performed to assess the degradation potential of the microbial species present in the oily sludge through augmentation, and using the augmented microbial inoculums to treat oily sludge in a slurry form containing mineral medium and water. The optimum pH of the slurry was found to be 8 for the biodegradation of oily sludge. The effect of oily sludge concentration, stirring rate, and treatment time on degradation of total petroleum hydrocarbons (TPH) was studied. It was found that the degradation process consists of two consecutive stages and that each stage follows a first-order kinetics. The first stage lasted 4 days followed by a second stage up to 7 days. The faster first stage had a rate constant of 0.1 day^?1, whereas the slower second stage had a rate constant of 0.056 day^?1. The kinetics was found to be time-dependent. The study showed that the mineral medium provided essential nutrients to the microbial species and that the degradation efficiency of the whole microbial species present in the oily sludge was quite high (～90%).     

Fate of Hydrocarbons During Oily Sludge Disposal in Soil

A 1,280-day laboratory simulation of the “landfarming” process explored the fate in soil of polynuclear aromatics (PNAs) and total extractable hydrocarbon residues originating from the disposal of an oily sludge. In addition to the measurement of CO₂ evolution, periodic analyses of PNAs and hydrocarbons monitored biodegradation activity. The estimation of carbon balance and of soil organic matter assessed the fate of residual hydrocarbons. Seven sludge applications during a 920-day active disposal period were followed by a 360-day inactive “closure” period with no further sludge applications. A burst of CO₂ evolution followed each sludge addition, but substantial amounts of undegraded hydrocarbons remained at the end of the study. Hydrocarbon accumulation did not inhibit biodegradation performance. Conversion of hydrocarbons to CO₂ predominated during active disposal; incorporation into soil organic matter predominated during the closure period. In this sludge, the predominant PNAs were degraded more completely (85%) than total hydrocarbons. Both biodegradation and abiotic losses of three- and four-ring PNAs contributed to this result. Some PNAs with five and six rings were more persistent, but these constituted only a small portion of the PNAs in the sludge. The study confirmed that the microbially mediated processes of mineralization and humification remove sludge hydrocarbons from soils of landfarms with reasonable efficiency.     

Degradation of mixtures of monochlorophenols and phenol as substrates for free and immobilized cells of Alcaligenes sp. A7-2

Summary The biodegradation of the three isomeric monochlorophenols 2-(2CP), 3- (3CP) and 4-chlorophenol (4CP) and phenol by the constructed strain Alcaligenes sp. A7-2 was investigated. Mineralization took place in the order: phenol >4CP >2CP >3CP, whereas 3CP was mineralized only co-metabolically. In substrate mixtures with phenol, degradation of 4CP was decelerated but degradation of 2CP was accelerated. Free cells in batch culture showed biphasic growth with an equimolar mixture of 2CP and 4CP as substrates, perhaps due to diauxie. Degradation patterns obtained with free cells in batch culture were confirmed with immobilized cells in continuous culture. Immobilized cells of Alcaligenes sp. A7-2 built up a biofilm on the lava that was used as filling material in the packed-bed reactors. The continuous cultures remained stable despite increasing input rates of chlorophenol and phenol mixtures up to 1.16 mMo1.1^–1.h^–1 for several weeks. Correspondence to: H.-J. Rehm     

Biodegradation of pyridine by freely suspended and immobilized Pimelobacter sp.

Freely suspended and Ca-alginate-immobilized cells of Pimelobacter sp. were used for degradation of pyridine. When the pyridine concentration was up to 2 g l^–1, freely suspended cells completely degraded pyridine regardless of the initial cell concentrations used. However, when the pyridine concentration increased to 4 g l^–1, the initial cell concentration in freely suspended cell culture should be higher than 1.5 g dry cell weight l^–1 for complete degradation of pyridine. In addition, a freely suspended cell culture with a high initial cell concentration resulted in a high volumetric pyridine-degradation rate, suggesting the potential use of immobilized cells for pyridine-degradation. When the immobilized cells were used for pyridine-degradation, neither specific pyridine-degradation rate nor tolerance against pyridine was improved. However, a high volumetric pyridine-degradation rate in the range 0.082–0.129 g l^–1 hr^–1 could be achieved by the immobilized cells because of the high cell concentration. Furthermore, when the immobilized cells were reused in degrading pyridine at a concentration of 2–4 g l^–1 they did not lose their pyridine-degrading activity for 2 weeks. Taken together, the data obtained here showed the feasibility of using immobilized cells for pyridine-degradation.     

Biodegradation potential of oily sludge by pure and mixed bacterial cultures

The biodegradation capacity of aliphatic and aromatic hydrocarbons of petrochemical oily sludge in liquid medium by a bacterial consortium and five pure bacterial cultures was analyzed. Three bacteria isolated from petrochemical oily sludge, identified as Stenotrophomonas acidaminiphila, Bacillus megaterium and Bacillus cibi, and two bacteria isolated from a soil contaminated by petrochemical waste, identified as Pseudomonas aeruginosa and Bacillus cereus demonstrated efficiency in oily sludge degradation when cultivated during 40 days. The bacterial consortium demonstrated an excellent oily sludge degradation capacity, reducing 90.7% of the aliphatic fraction and 51.8% of the aromatic fraction, as well as biosurfactant production capacity, achieving 39.4% reduction of surface tension of the culture medium and an emulsifying activity of 55.1%. The results indicated that the bacterial consortium has potential to be applied in bioremediation of petrochemical oily sludge contaminated environments, favoring the reduction of environmental passives and increasing industrial productivity.     

Biodegradation of Naphthalene by Pseudomonas stutzeri in Marine Environments: Testing Cells Entrapment in Calcium Alginate for Use in Water Detoxification

The biodegradation of naphthalene in sea water by freely suspended and alginate-entrapped cells of Pseudomonas stutzeri 19SMN4 has been investigated in batch cultures. The results showed that immobilized cells can be stored at 4°C for 1 month without loss of viability. The biodegradation was highly affected by the availability of nitrogen and phosphorous, so at 30°C a naphthalene concentration of 25 mM was almost completely degraded (93%) by free cells in 6 days in samples supplemented with these nutrients, whereas only 42% naphthalene was consumed in the nonsupplemented samples. Biodegradation was much slower at 16°C than at 30°C; after 6 days of culture at 30°C, almost all naphthalene was degradated by free and immobilized cells, whereas only 22% and 34% at 16°C, respectively. The degradation rate remained unaffected when the naphthalene concentration was reduced from 25 to 10 mM. Alginate of three different viscosities was used for immobilization of cells. After 7 days of culture, beads formed with 31.4 cP alginate were fragmented, whereas beads formed with 240 and 3600 cP did not display structural changes and afforded the same degradation rate. Beads formed with high-viscosity alginate retained cells more efficiently.     

Production ofLactococcus lactis biomass by immobilized cell technology

Summary Calcium alginate beads containingLactococcus lactis cells were used for three batch fermentations of milk or a commercially available growth medium (Gold Complete, Nordica) with the aim of producing concentrated cultures. Repeated fermentations did not significantly increase bead CFU counts which were between 3.3–7.8×10¹⁰ CFU/g. During the second and third fermentations, which lasted 6 h each, the bead populations decreased if the incubation was extended over 2 h. There was cell release from the beads. Fermentation media and fermentation time all had an effect on free cell counts, but none of these factors statistically interacted. Free cell counts were higher at the end of fermentations 2 and 3 than in the first fermentation and approximately 50% of the population was in the free state. Free cell counts were higher when the beads were incubated in Gold complete than in milk. Although the total bacterial population of a standard free cell fermentation was always higher than those having immobilized cells, immobilized cell technology did enable the production of dense cultures.     

The mechanism of cadmium removal from aqueous solution by nonmetabolizing free and immobilized live biomass of Rhizopus oligosporus

A preliminary study on the removal of cadmium by nonmetabolizing live biomass of Rhizopus oligosporus from aqueous solution is presented. The equilibrium of the process was in all cases well described by the Langmuir sorption isotherm, suggesting that the process was a chemical, equilibrated and saturable mechanism which reflected the predominantly site-specific mechanism on the cell surface. A curve of Scatchard transformation plots reflected the covalent nature of Cd²⁺ adsorption by the cells. The maximum cadmium uptake capacities were 34.25 mg/g for immobilized cells and 17.09 mg/g for free cells. Some factorial experiments in shake flasks were performed in order to investigate the effect of different initial cadmium concentrations and biomass concentrations on the equilibrium. Experimental results showed a reverse trend of the influence of the immobilized and free biomass concentration on the cadmium specific uptake capacity. The immobilized cells had a higher specific cadmium uptake capacity with increasing biomass concentrations compared to free cells. In a bioreactor, the cadmium uptake capacity of immobilized cells (q_max = 30.1–37.5 mg/g) was similar to that observed in shake flask experiments (q_max = 34.25 mg/g) whereas with free cells the bioreactor q_max of 4.8–13.0 mg/g; was much lower than in shake flasks (q_max = 17.09 mg/g), suggesting that cadmium biosorption by immobilized cells of R. oligosporus might be further improved in bigger reactors. EDAX and transmission electron microscopic experiments on the fungal biomass indicated that the presence of Cd²⁺ sequestrated to the cell wall was due to bioadsorption.     

Methyl mercury uptake by free and immobilized cyanobacterium

Methyl mercury uptake in free cells and different immobilizates of the cyanobacteriumNostoc calcicola has been examined. The general growth of the immobilized cyanobacterial cells could be negatively correlated with methyl mercury uptake. Alginate spheres proved most efficient in terms of uptake rate (0.48 nmol mg protein^–1 min^–1, 10 min) and total bioaccumulation (10.71 nmol mg protein^–1, 1 h) with a bioconcentration factor of 3.3×10³. Alginate biofilms showed a faster methyl mercury accumulation rate (0.83 nmol mg protein^–1 min^–1, 10 min) with a saturation of 10.28 nmol mg protein^–1 reached within only 30 min (bioconcentration factor, 3.1×10³). Foam preparations with a slow initial uptake approximated biofilms but were characterized by a lower bioconcentration factor (2.8×10³). Free cells, in comparison, maintained the initial slow rate of uptake (0.62 nmol mg protein^–1 min^–1, 10 min), saturating at 30 min (8.81 nmol mg protein^–1), and the resultant lowest bioconcentration factor (2.7×10³). Cell ageing (30 days) brought a drastic reduction (3-fold) in organomercury uptake by free cells while alginate spheres maintained the same potential. Foam preparations of the same age showed a significant improvement in methyl mercury uptake followed by only a marginal decline in alginate biofilms. Data are discussed in the light of the physiological efficiency and longevity of immobilized cells.     

Degradation of polycyclic aromatic hydrocarbons by an immobilized mixed bacterial culture

The mixed bacterial culture MK1 was capable of degrading a wide spectrum of aromatic compounds both as free and as immobilized cells. By offering anthracene oil or a defined mixture of phenol, naphthalene, phenanthrene, anthracene and pyrene (in concentrations of 0.1–0.2 mm, respectively) as sources of carbon and energy, a specific degradation pattern correlating with the condensation degree was observed. Regarding the defined mixture of aromatic hydrocarbons, complete metabolism was reached for phenol (0.1 mm) after 1 day, for naphthalene (0.1 mm) after 2 days and for phenanthrene (0.1 mm) after 15 days of cultivation. The conversion of anthracene (0.1 mm) and pyrene (0.1 mm) resulted in minimal residual concentrations, analogous to fluoranthene and pyrene of the anthracene oil (0.1%). Maximal total degradation for the tricyclic compounds dibenzofurane, fluorene, dibenzothiophene, phenanthrene and anthracene of the anthracene oil (0.1%) occurred after 5 days. In general, a significant metabolisation of the tetracyclic aromatic hydrocarbons fluoranthene and pyrene was observed after the degradation of phenol, naphthalene and most of the tricyclic compounds. Doubling the start concentrations of the polycyclic aromatic hydrocarbons effected higher degradation rates. Cell growth occurred simultaneously with the conversion of phenol, naphthalene and the tricyclic compounds. The immobilized cells showed stable growth and, compared to freely suspended cells, the same degradation sequence as well as an equivalent degradation potential — even in a model soil system. Correspondence to: I. Wiesel     

Production of cellulase by Aspergillus niger biofilms developed on polyester cloth

AIMS: To compare cellulase production by Aspergillus niger ATCC 10864 biofilms on polyester cloth and freely suspended cultures in shaken flasks and microbioreactors of bubble column type. METHODS AND RESULTS: Both shaken flasks and oxygenated microbioreactors containing 40 ml of production medium were used to compare cellulase secretion by free mycelium and biofilm cultures. Free mycelium cultures grew better in flasks than in microbioreactors producing compact and fluffy pellets, respectively, while the opposite was found for biofilm cultures without any visible change in biofilm morphology. Cellulase activities and volumetric productivities attained by biofilms in flask cultures were 70% higher than that produced by free mycelium cultures and threefold higher when biofilms were grown in microbioreactors. CONCLUSIONS: Fungal biofilms developed on polyester cloth in both flasks and microbioreactors produce higher cellulase yields and volumetric productivities than free mycelium cultures at lower biomass levels. SIGNIFICANCE AND IMPACT OF THE STUDY: The results of the present study are of commercial and biological interest. All productivity parameters revealed that fungal biofilms may be used for the production of cellulase and other proteins in various types of bioreactors. Moreover, they may be used as model systems to study differential gene expression related to cell adhesion.     

Organic Bulking Agents for Enhancing Oil Bioremediation in Soil

Soil contaminated with oil is bioremediated by optimizing conditions for microbial activity. Often the question arises about the benefits of bulking with organic materials to improve soil conditions to enhance degradation of the less biodegradable or less bioavailable components. An investigation was undertaken in the laboratory with the objective of measuring the influence of bulking with dried plant material, bermudagrass, and alfalfa on the degradation of oily sludge added to soil. The oily sludge was diluted 50:50 on a weight basis with soil to achieve a final concentration of 100 g oil and grease kg^-1 of final soil mixture. Bulking agents were added 40 d after dilution of the sludge and optimization of environmental conditions to allow time for the readily decomposable fraction to be degraded before amendment with bulking agents. Populations of heterotrophic microorganisms increased approximately ten times by 40 and 80 d after addition of bulking agents, but the numbers of hydrocarbon-degrading microorganisms did not significantly increase above the number in the nonbulked control. Bulking agents increased the quantity of total petroleum hydrocarbons degraded by approximately 20% during the first 40 d after being added. Disappearance of hydrocarbons for bulked treatments was much slower during the next 40 d, such that the total petroleum hydrocarbon content for both bulked and nonbulked treatments generally was not significantly different at the end. It appears that adding bulking agents may enhance the rate of decomposition of total petroleum hydrocarbons by stimulating the general heterotrophic population of microorganisms, but the influence may not be sustained to influence the extent of decomposition.     

Comparison of bio-augmentation and composting for remediation of oily sludge: A field-scale study in China

Two bioremediation technologies were performed in order to explore a better treatment process for an oily sludge restoration in China during 2004. The bioremediation by augmentation of biopreparation was compared with a conventional composting. The oily sludge and oil-polluted soil were received from an oil production plant. The total hydrocarbon content (THC) varied from 327.7 to 371.2 g kg⁻¹ of dry sludge and the THC in contaminated soil was 151.0 g kg⁻¹. Before application of preparation, straw, sawdust, top sand and pure soil were added in different proportions to the sludge and soil and mixed thoroughly. Such sludge and soil composites were used for negative controls and for activation of indigenous oil degrading microorganisms with addition of fertilizer (positive controls). For composting, crude manure and straw were added to the oily sludge and the THC was 101.4 g kg⁻¹. The biopreparation was applied every 2 weeks and experiment lasted 56 days under the ambient temperature. The sludge was mixed and watered every 3 days. After three times of biopreparation application, the THC decreased by 46–53% in the oily sludge and soil, while in the positive controls (activation of indigenous microorganisms) the THC decreased by 13–23%, and there was no oil degradation in negative controls After composting, the THC decreased by 31% in the oily sludge. The planting of Tall Fescue (Festuca arundinace) revealed a decrease of sludge toxicity after application of both bioremediation technologies and additionally decreased the THC by 5–7%.     

Degradation of naphthalene by cells of Pseudomonas sp. strain NGK 1 immobilized in alginate, agar and polyacrylamide

A Pseudomonas sp. strain NGK 1 (NCIM 5120) was immobilized in various matrices, namely, alginate, agar (1.8 × 10¹¹ cfu g⁻¹ beads) and polyacrylamide (1.6 × 10¹¹ cfu g⁻¹ beads). The degradation of naphthalene was studied, by freely suspended cells (4 × 10¹⁰ cfu ml⁻¹) and immobilized cells in batches, with shaken culture and continuous degradation in a packed-bed reactor. Free cells brought about the complete degradation of 25 mmol naphthalene after 3 days of incubation, whereas, a maximum of 30 mmol naphthalene was degraded by the bacteria after 3–4 days of incubation with 50 mmol and 75 mmol naphthalene, and no further degradation was observed even after 15 days of incubation. Alginate-entrapped cells had degraded 25 mmol naphthalene after 3.5 days of incubation, whereas agar- and polyacrylamide-entrapped cells took 2.5 days; 50 mmol naphthalene was completely degraded by the immobilized cells after 6–7 days of incubation. Maximum amounts of 55 mmol, 70 mmol and 67 mmol naphthalene were degraded, from an initial 75 mmol naphthalene, by the alginate-, agar- and polyacrylamide-entrapped cells after 15 days of incubation. When the cell concentrations were doubled, 25 mmol and 50 mmol naphthalene were degraded after 2 and 5.5 days of incubation by the immobilized cells. Complete degradation of 75 mmol naphthalene occurred after 10 days incubation with agar- and polyacrylamide-entrapped␣cells, whereas only 60 mmol naphthalene was degraded by alginate-entrapped cells after 15 days of␣incubation. Further, with 25 mmol naphthalene, alginate-, agar- and polyacrylamide-entrapped cells (1.8 × 10¹¹ cfu g⁻¹ beads) could be reused 18, 12 and 23 times respectively. During continuous degradation in a packed-bed reactor, 80 mmol naphthalene 100 ml⁻¹ h⁻¹ was degraded by alginate- and polyacrylamide-entrapped cells whereas 80 mmol naphthalene 125 ml⁻¹␣h⁻¹ was degraded by agar-entrapped cells. Received: 21 October 1997 / Received revision: 15 January 1998 / Accepted: 18 January 1998