Monitoring of the Degradation Activities and the Diversity of the Microbial Community Degrading Refinery Waste Sludge

The biodegradation of n-alkanes and branched alkanes from waste sludge were observed in landfarming soils of Motor Oil Hellas (a petroleum refinery) and changes in the bacterial communities in the soils were monitored during the remediation. Bacterial 16S rRNA gene (rDNA)-based community fingerprint patterns were obtained from soil samples by terminal restriction fragment length polymorphism (T-RFLP) analysis. Changes in T-RFLP fingerprints, as well as in the petroleum contaminant composition of the polluted soil, correlated with degradation activities in field tests.     

大气粉尘中常见多环芳烃类化合物的提取及含量测定

采用便携式PM2.5采样器和玻璃纤维滤膜对大气粉尘进行采样,以加压液体萃取、超声波辅助提取和微波辅助提取3种方法对多环芳烃类化合物进行提取,以超高效液相色谱和质谱联用方法进行测定。共检测出16种多环芳烃类化合物,分别为:萘、苊、苊烯、芴、菲、蒽、荧蒽、芘、苯并[a]蒽、■、苯并[b]荧蒽、苯并[k]荧蒽、苯并[a]芘、二苯并[a,h]蒽、茚并[1,2,3-c,d]芘和苯并[g,h,i]苝。3种提取方法均可有效提取大气粉尘中的多环芳烃类化合物,加压液体萃取法的提取效率最高、超声波辅助提取法次之、微波辅助提取法最低。超高效液相色谱和质谱连用方法可有效地分析大气粉尘中的多环芳烃类化合物,线性方程的相关系数不低于0.998 3,测定值的相对标准偏差不超过0.80%,检出限不高于0.27 ng/g。     

Pseudomonas sp. (Strain 10–1B): A potential inoculum candidate for green and sustainable remediation

Soil pollution caused by polycyclic aromatic hydrocarbons (PAHs) is a consequence of various industrial processes which destabilizes the ecosystem. Bioremediation by bacteria is a cost‐effective and environmentally safe solution for reducing or eliminating pollutants in soils. In the present study, we artificially polluted agricultural soil with used automobile engine oil with a high PAH content and then isolated bacteria from the soil after 10 weeks. Pseudomonas sp. strain 10–1B was isolated from the bacterial community that endured this artificial pollution. We sequenced its genomic DNA on Illumina MiSeq sequencer and evaluated its ability to solubilize phosphate, fix atmospheric nitrogen, and produce indoleacetic acid, in vitro, to ascertain its potential for contribution to soil fertility. Its genome annotation predicted several dioxygenases, reductases, ferredoxin, and Rieske proteins important in the ring hydroxylation initiating PAH degradation. The strain was positive for the soil fertility attributes evaluated. Such combination of attributes is important for any potential bacterium partaking in sustainable bioremediation of PAH‐polluted soil.     

Use of fungal technology in soil remediation: A Case Study

Two white rot fungi Irpex lacteus and Pleurotus ostreatus and a PAH-degrading bacterial strain of Pseudomonas putida were used as inoculum for bioremediation of petroleum hydrocarbon-contaminated soil from a manufactured-gas-plant-area. Also two cocultures comprising a fungus with Pseudomonas putida were applied. After 10-week treatment out of 12 different PAHs, concentration of phenanthrene, anthracene, fluoranthene and pyrene decreased up to 66%. The ecotoxicity of the soil after bioremediation did not reveal any effect on the survival of Daphnia magna, a crustacian. However, the toxic effect on seed germination of plant Brassica alba and oxidoreductase activity of bacterium Bacillus cereus decreased after 5 and 10 weeks of treatment.     

Effects of physicochemical factors on PAH degradation by Planomicrobium alkanoclasticum

Polycyclic aromatic hydrocarbons (PAHs) are environmental pollutants that are mutagenic, carcinogenic, and toxic to living organisms. Here, the ability and effectiveness of selected bacteria isolated from an oil‐contaminated area in biodegrading PAHs were evaluated, and the optimal conditions conducive to bacterial PAH biodegradation were determined. Of six bacterial isolates identified based on their 16S rRNA sequences, Planomicrobium alkanoclasticum could subsist on and consume nearly all hydrocarbons according to the 2,6‐dichlorophenolindophenol assay. The efficacy of this isolate at PAH biodegradation was then empirically confirmed. After 30 days of incubation, P. alkanoclasticum degraded 90.8% of the 16 PAH compounds analyzed and fully degraded eight of them. The optimum P. alkanoclasticum growth conditions were 35°C, pH 7.5, and NaNO₃ as the nitrogen source. Under these biostimulant conditions, P. alkanoclasticum degraded 91.4% of the total PAH concentration and completely decomposed seven PAHs after 15 days incubation. Hence, P. alkanoclasticum is an apt candidate for the biodegradation of PAHs and the bioremediation of sites contaminated by them.     

Characterisation of polycyclic aromatic hydrocarbons in flue gas and residues of a full scale fluidized bed combustor combusting non-hazardous industrial waste

This paper studies the fate of PAHs in full scale incinerators by analysing the concentration of the 16 EPA-PAHs in both the input waste and all the outputs of a full scale Fluidized Bed Combustor (FBC). Of the analysed waste inputs i.e. Waste Water Treatment (WWT) sludge, Refuse Derived Fuel (RDF) and Automotive Shredder Residue (ASR), RDF and ASR were the main PAH sources, with phenanthrene, fluoranthene and pyrene being the most important PAHs. In the flue gas sampled at the stack, naphthalene was the only predominant PAH, indicating that the PAHs in FBC’s combustion gas were newly formed and did not remain from the input waste. Of the other outputs, the boiler and fly ash contained no detectable levels of PAHs, whereas the flue gas cleaning residue contained only low concentrations of naphthalene, probably adsorbed from the flue gas. The PAH fingerprint of the bottom ash corresponded rather well to the PAH fingerprint of the RDF and ASR, indicating that the PAHs in this output, in contrast to the other outputs, were mainly remainders from the PAHs in the waste inputs. A PAH mass balance showed that the total PAH input/output ratio of the FBC ranged from about 100 to about 2600 depending on the waste input composition and the obtained combustion conditions. In all cases, the FBC was clearly a net PAH sink.     

Remediation of Soil and Ground Water Contaminated with PAH using Heat and Fe(II)-EDTA Catalyzed Persulfate Oxidation

The feasibility of degrading 16 USEPA priority polycyclic aromatic (PAH) hydrocarbons (PAHs) with heat and Fe(II)-EDTA catalyzed persulfate oxidation was investigated in the laboratory. The experiments were conducted to determine the effects of temperature (i.e. 20 ^∘C, 30 ^∘C and 40 ^∘ C) and iron-chelate levels (i.e., 250 mg/L-, 375 mg/L- and 500 mg/L-Fe(II)) on the degradation of dissolved PAHs in aqueous systems, using a series of amber glass jars as the reactors that were placed on a shaker inside an incubator for temperature control. Each experiment was run in duplicate and had two controls (i.e., no persulfate in systems). Samples were collected after a reaction period of 144 hrs and measured for PAHs, pH and sodium persulfate levels. The extent of degradation of PAHs was determined by comparing the data for samples with the controls. The experimental results showed that persulfate oxidation under each of the tested conditions effectively degraded the 16 target PAHs. All of the targeted PAHs were degraded to below the instrument detection limits (∼4 μ/L) from a range of initial concentration (i.e., 5 μ/L for benzo(a)pyrene to 57 μ/L for Phenanthrene) within 144 hrs with 5 g/L of sodium persulfate at 20 ^∘ C, 30 ^∘C and 40 ^∘C. The data indicated that the persulfate oxidation was effective in degrading the PAHs and that external heat and iron catalysts might not be needed for the degradation of PAHs. The Fe(II)-EDTA catalyzed persulfate also effectively degraded PAHs in the study. In addition, the data on the variation of persulfate concentrations during the experiments indicated that Fe(II)-EDTA accelerated the consumption of persulfate ions. The obtained degradation data cannot be used to evaluate the influence of temperature and Fe(II) levels on the PAH degradation because the PAHs under each of the tested conditions were degraded to below the instrument detection limit within the first sampling point. However, these experiments have demonstrated the feasibility of degrading PAHs in aqueous systems with persulfate oxidation. Additional tests are being conducted to evaluate the effectiveness of treating PAHs in soils and obtaining the rate of degradation of PAHs with persulfate oxidation. Two sets of laboratory experiments were conducted to evaluate the ability of sodium persulfate in oxidizing real world PAH-contaminated soils collected from a Superfund site in Connecticut. The first set of soil sample were treated only with persulfate and to the second batch, mixture of persulfate and Fe(II)-EDTA solutions were added. The results of the second test showed that within 24 hours, 75% to 100% of the initial concentrations of seven PAH compounds detected in the soil samples were degraded by sodium persulfate mixed with FE(II)-EDTA.     

In Situ flushing of contaminated soils from a refinery: Organic compounds and metal removals

This article demonstrates the applicability of in situ flushing for the remediation of soil contaminated with petroleum hydrocarbons at a Mexican refinery. The initial average total petroleum hydrocarbon (TPH) concentration for the demonstration field test was 55,156 g/kg. After six weeks of in situ flushing with alternate periods of water and water/surfactant, an average concentration of 1,407 mg/kg was reached, achieving a total removal efficiency of 98 percent. At the end of the process, no hydrocarbons such as diesel; gasoline; benzene, toluene, ethyl benzene, and xylene (BTEX); or petroleum aromatic hydrocarbons (PAHs) were found. Iron washing achieved a removal efficiency of 70 percent, and for vanadium, the removal efficiency was 94.4 percent. The volume of soil treated was 41.6 m³ (38 m²), equivalent to 69.5 tons of soil. A rough calculation of the process costs estimated a total cost of $104.20/m³ ($114.00/m²). Our research indicates that there are a few studies demonstrating in situ flushing experiences under field conditions where both organic (TPH, diesel, gasoline, PAHs, BTEX) and metal (iron and vanadium) removals are reported. © 2004 Wiley Periodicals, Inc.     

Ethanol washing of PAH-contaminated soil and Fenton oxidation of washing solution

As a means to remediate soil contaminated by polycyclic aromatic hydrocarbons, we investigated a combined process involving ethanol washing followed by a Fenton oxidation reaction. Artificial loamy soil was contaminated with various representative polycyclic aromatic hydrocarbons (i.e., fluorene, anthracene, pyrene, benzo(b)fluoranthene, or benzo(a)pyrene) at concentrations ten times higher than regulatory soil standards of The Netherlands or Canada, and then washed four times in ethanol, which reduced the concentration of polycyclic aromatic hydrocarbon contamination to below the regulatory standard. Fenton oxidation of ethanol solutions containing anthracene, benzo(a)pyrene, pyrene, acenaphthylene, acenaphthene, benz(a)anthracene, benzo(j)fluoranthene, or indeno(1,2,3-cd)pyrene showed a removal efficiency of 73.3%–99.0%; by contrast, solutions containing naphthalene, fluorene, fluoranthene, phenanthrene, or benzo(b)fluoranthene showed a removal efficiency of 9.6%–27.6%. Since each of the nonremediated polycyclic aromatic hydrocarbons, excluding benzo(b)fluoranthene, are easily biodegradable, these results indicate that the proposed treatment can be successfully applied to polycyclic aromatic hydrocarbon-contaminated soil that does not contain high concentrations of benzo(b)fluoranthene. The main reaction products resulting from Fenton oxidation of ethanol solutions containing anthracene or benz(a)anthracene were anthraquinon or benz(a)anthracene-7,12-dione, respectively; while 1,8-naphthalic anhydride was produced by solutions of acenaphthylene and acenaphthene, and 9-fluorenone by a fluorene solution. Received: June 9, 1998 / Accepted: March 24, 1999     

Microbial communities and greenhouse gas emissions associated with the biodegradation of specified risk material in compost

Provided that infectious prions (PrP^Sc) are inactivated, composting of specified risk material (SRM) may be a viable alternative to rendering and landfilling. In this study, bacterial and fungal communities as well as greenhouse gas emissions associated with the degradation of SRM were examined in laboratory composters over two 14 day composting cycles. Chicken feathers were mixed into compost to enrich for microbial communities involved in the degradation of keratin and other recalcitrant proteins such as prions. Feathers altered the composition of bacterial and fungal communities primarily during the first cycle. The bacterial genera Saccharomonospora, Thermobifida, Thermoactinomycetaceae, Thiohalospira, Pseudomonas, Actinomadura, and Enterobacter, and the fungal genera Dothideomycetes, Cladosporium, Chaetomium, and Trichaptum were identified as candidates involved in SRM degradation. Feathers increased (P < 0.05) headspace concentrations of CH₄ primarily during the early stages of the first cycle and N₂O during the second. Although inclusion of feathers in compost increases greenhouse gas emissions, it may promote the establishment of microbial communities that are more adept at degrading SRM and recalcitrant proteins such as keratin and PrP^Sc.     

Biofilm mediated degradation of commercially available LDPE films by bacterial strains isolated from partially degraded plastic

Biodegradation is an attractive approach for the elimination of synthetic polymers, pervasively accumulated in natural environments and generating ecological problems. The present work investigated the degradation of low‐density polyethylene (PE) by three Bacillus sp., that is, ISJ36, ISJ38, and ISJ40. The degree of biodegradation was assessed by measuring hydrophobicity, viability, and total protein content of bacterial biofilm attached to the PE surface. Although all three bacterial strains were able to establish an active biofilm community on the PE surface, ISJ40 showed better affinity toward PE degradation than the other two. Bacterial colonization and physical changes on the PE surface were visualized by scanning electron microscopy. Fourier transform infrared spectroscopy analysis revealed alteration in the intensities of functional groups along with an increase in the carbonyl bond indexes. The study results suggest that the Bacillus strain ISJ40 can be used as a potential degrader for the eco‐friendly treatment of PE waste.     

Influence of pH and C/N Ratio on Poly(Vinyl Alcohol) Biodegradation in Mixed Bacterial Culture

Cultivation conditions affecting poly(vinyl alcohol) (PVA) degradation by a mixed bacterial culture of Bacillus sp. and Curtobacterium sp. were investigated. Bacterial strains used in this study were isolated from the watercourse and the sewage sludge of vinylonfibre mill by enrichments on PVA as the sole carbon source. The results showed that PVA was greatly degraded under the following conditions: 0.5% PVA as a substrate at the initial medium pH of 8 with 0.15% glucose and urea at C/U ratio 1.5:1 and 1% bacterial inoculum, at a temperature of 35 °C and a shaking speed of 110 rpm. The analysis of FTIR and ¹H NMR spectra before and after biodegradation indicate fission of the PVA molecular chain during the incubation.     

Lysinibacillus sphaericus and Geobacillus sp Biodegradation of Petroleum Hydrocarbons and Biosurfactant Production

Petroleum oil is a major driver of worldwide economic activity, but it has also created contamination problems during the storage and refining process. Also, unconventional resources are natural resources, which require greater than industry‐standard levels of technology or investment to exploit. In the case of unconventional hydrocarbon resources, additional technology, energy, and capital have to be applied to extract the gas or oil. Bioremediation of petroleum spill is considered of great importance due to the contaminating effects on human health and the environment. For this reason, it is important to reduce total petroleum hydrocarbons (TPH) in contaminated soil. In addition, biosurfactant production is a desirable property of hydrocarbon‐degrading microorganisms. Seven strains belonging to Lysinibacillus sphaericus and Geobacillus sp were selected to evaluate their ability to biodegrade TPH in the presence of toxic metals, their potential to produce biosurfactants, and their ability to improve the biodegradation rate. The seven bacterial strains examined in this study were able to utilize crude petroleum‐oil hydrocarbons as the sole source of carbon and energy. In addition, their ability to degrade crude oil was not affected by the presence of toxic metals such as chromium and arsenic. At the same time, the strains were able to reduce toxic metals concentration through biosorption processes. Biosurfactant production was determined using the drop‐collapsed method for all strains, and they were characterized as both anionic and cationic biosurfactants. Biosurfactants showed an increase in biodegradation efficiency both in liquid minimal salt medium and landfarming treatments. The final results in field tests showed an efficiency of 93 percent reduction in crude oil concentration by the selected consortium compared to soil without consortium. The authors propose L. sphaericus and Geobacillus sp consortium as an optimum treatment for contaminated soils. In addition, production of biosurfactants could have an application in the extraction of crude oil from unconventional hydrocarbon resources. © 2014 Wiley Periodicals, Inc.     

Biochemical and Genetic Characterization of an Extracellular Poly(3-Hydroxybutyrate) Depolymerase from Acidovorax Sp. Strain TP4

To determine the properties of enzymes from bacteria that degrade polypropiolactone (PPL), we isolated 13 PPL-degrading bacteria from pond water, river water, and soil. Nine of these strains were identified as Acidovorax sp., three as Variovorax paradoxus, and one as Sphingomonas paucimobilis. All the isolates also degraded poly(3-hydroxybutyrate) (PHB). A PPL-degrading enzyme was purified to electrophoretical homogeneity from one of these bacteria, designated Acidovorax sp. TP4. The purified enzyme also degraded PHB. The molecular weight of the enzyme was estimated as about 50,000. The enzyme activity was inhibited by diisopropylfluorophosphate, dithiothreitol, and Triton X-100. The structural gene of the depolymerase was cloned in Escherichia coli. The nucleotide sequence of the cloned DNA fragment contained an open reading frame (1476 bp) specifying a protein with a deduced molecular weight of 50,961 (491 amino acids). The deduced overall sequence was very similar to that of a PHB depolymerase of Comamonas acidovorans YM1609. From these results it was concluded that the isolated PPL-degrading enzyme belongs to the class of PHB depolymerases. A conserved amino acid sequence, Gly-X₁-Ser-X₂-Gly (lipase box), was found at the N-terminal side of the amino acid sequence. Site-directed mutagenesis of the TP4 enzyme confirmed that ²⁰Ser in the lipase box was essential for the enzyme activity. This is the first report of the isolation a PHB depolymerase from Acidovorax.     

Bacterial Poly(Hydroxyalkanoate) Polymer Production from the Biodiesel Co-product Stream

A co-product stream from soy-based biodiesel production (CSBP) containing glycerol, fatty acid soaps, and residual fatty acid methyl esters (FAME) was utilized as a fermentation feedstock for the bacterial synthesis of poly(3-hydroxybutyrate) (PHB) and medium-chain-length poly(hydroxyalkanoate) (mcl-PHA) polymers. Pseudomonas oleovorans NRRL B-14682 and P. corrugata 388 grew and synthesized PHB and mcl-PHA, respectively, when cultivated in up to 5% (w/v) CSBP. In shake flask culture, P. oleovorans grew to 1.3 ± 0.1 g/L (PHA cellular productivity = 13–27% of the bacterial cell dry weight; CDW) regardless of the initial CSBP concentration, whereas P. corrugata reached maximum cell yields of 2.1 g/L at 1% CSBP, which tapered off to 1.7 g/L as the CSBP media concentration was increased to 5% (maximum PHA cellular productivity = 42% of the CDW at 3% CSBP). While P. oleovorans synthesized PHB from CSBP, P. corrugata produced mcl-PHA consisting primarily of 3-hydroxyoctanoic acid (C_8:0; 39 ± 2 mol%), 3-hydroxydecanoic acid (C_10:0; 26 ± 2 mol%) and 3-hydroxytetradecadienoic acid (C_14:2; 15 ± 1 mol%). The molar mass (M_n) of the PHB polymer decreased by 53% as the initial CSBP culture concentration was increased from 1% to 5% (w/v). In contrast, the M_n of the mcl-PHA polymer produced by P. corrugata remained constant over the range of CSBP concentrations used.     

Comparing the Growth Characteristics of Three Bacteria Involved in Degrading Rubbers

Three bacteria isolated for degradation of rubbers were compared for their growth characteristics derived from the Bradford protein assay and turbidity (optical density, OD) measurement. Both Alcaligenes xylosoxidans T2 and Pseudomonas aeruginosa GP10 were fast-growing bacteria while Nocardia corynebacterioides S3 was a slow grower utilizing rubber as the sole source of carbon and energy, but the extent of degradation was lower by the formers than the latter. A. xylosoxidans T2, P. aeruginosa GP10 and N. corynebacterioides S3 showed a typical sigmoidal growth pattern based on binding of Coomassie Brilliant Blue G250 to bacterial proteins and spectrophotometrical measurement at 600 nm. Both assays showed similar growth characteristics for all three bacteria in this study. Degradation of rubber was more pronounced by N. corynebacterioides S3 than either A. xylosoxidans T2 or P. aeruginosa GP10 during 70 days of incubation. Our results suggest that slow-growing bacteria may play a much greater role in degrading polymeric materials than was previously believed.     

Quantification of Polymerisation Processes During The Oxidative Degradation of 14C-Labelled Chlorophenols

In experiments employing the lignocellulose-decaying basidiomycetes Trametes versicolor and Stropharia rugosoannulata degrading uniformly¹⁴C-labelled 2,4-dichlorophenol and pentachlorophenol, acombination of size exclusion chromatography (SEC),fractionation, and -scintillation counting wasapplied to quantify polymerisation products formed duringchlorophenol degradation. Time-dependent mass balances weregenerated by analysis of ¹⁴C in polymerisation products,CO₂, as well as monomer non-polar and polar metabolites.Approximately 30% of the chlorophenols were found to bepolymerised. A major fraction of the polymerised productscorresponded to a molecular weight range from 0.24 – 40 kDa.Only a minor fraction could be attributed to a molecularweight >40 kDa. This method proved to be useful inquantification of polymerisation products and kinetics of thepolymerisation processes, whereas UV/Vis detection ofpolymerisation products separated by SEC led to false positiveresults. The SEC-¹⁴C method could also be applied forother complex processes where polymerisation ordepolymerisation occurs (humification, degradation oflignocellulose, formation of bound residues from xenobioticssuch as polycyclic aromatic hydrocarbons or 2,4,6-trinitrotoluene) and where spectrophotometric determinationsare difficult or impossible.     

Synthesis of copolyesters containing poly(ethylene terephthalate) and poly(ε-caprolactone) units and their susceptibility toPseudomonas sp. lipase

Copolyesters containing poly(ethylene terephthalate) (PET) and poly(-caprolactone) (PCL) were synthesized from PET and PCL homopolymers by transesterification reaction at 270°C in the presence of catalyst. The copolyesters were characterized by¹³C-NMR and differential scanning calorimetry (DSC). The degradation behavior of PCL byPseudomonas sp. lipase in buffer solution (pH 7) and tetrahydrofuran (THF) was investigated by gel permeation chromatography (GPC) and¹H-NMR. From these experiments, it was found thatPseudomonas sp. lipase acted endoenzymatically on PCL. Using this lipase, degradation tests for PET/PCL copolyesters whose PCL content was below 50% by weight were also performed in buffer solution (pH 7). However, evenPseudomonas sp. lipase with high degradation activity on PCL did not easily degrade the PCL unit in PET/PCL copolyesters.     

Degradation of Poly(3-hydroxybutyrate) and Poly(3-hydroxybutyrate-co-3-hydroxyvalerate) by Some Soil Aspergillus spp.

Systematic screening of 45 soil fungi for degradation polyhydroxyalkanoic acids (PHAs) has led to the selection of 6 potent Aspergillus isolates belonging to A. flavus, A. oryzae, A. parasiticus, and A. racemosus. Degradation of PHAs as determined by tube assay method revealed that these Aspergillus spp. were more efficient in degrading poly(3-hydroxybutyrate) [P(3HB)] compared to copolymer of 3-hydroxybutyric acid and 3-hydroxyvaleric acid (P3HB-co-16% 3HV). Moreover, the extent of degradation in mineral base medium was much better than those in complex organic medium. For all the Aspergillus spp. tested, maximum degradation was recorded at a temperature of 37°C with significant inhibition of growth. The optimum pH range for degradation was 6.5–7.0 with degradation being maximum at pH 6.8. The extent of polymer degradation increased with increase in substrate concentration, the optimum concentration for most of the cultures being 0.4% and 0.2% (w/v) for P(3HB) and P(3HB-co-16%3HV) respectively. Supplementation of the degradation medium with additional carbon sources exerted significant inhibitory effect on both P(3HB) and P(3HB-co-16%3HV) degradation.     

Degradation of poly(3-hydroxybutyrate), PHB,by bacteria and purification of a novel PHB depolymerase fromComamonas sp.

Bacteria capable of growing on poly(3-hydroxybutyrate), PHB, as the sole source of carbon and energy were isolated from various soils, lake water, activated sludge, and air. Although all bacteria utilized a wide variety of monomeric substrates for growth, most of the strains were restricted to degrade PHB and copolymers of 3-hydroxybutyrate and 3-hydroxyvalerate, P(3HB-co-3HV). Five strains were also able to decompose a homopolymer of 3-hydroxyvalerate, PHV. Poly(3-hydroxyoctanoate), PHO, was not degraded by any of the isolates. One strain, which was identified asComamonas sp., was selected, and the extracellular depolymerase of this strain was purified from the medium by ammonium sulfate precipitation and by chromatography on DEAE-Sephacel and Butyl-Sepharose 4B. The purified PHB depolymerase was not a glycoprotein. The relative molecular masses of the native enzyme and of the subunits were 45,000 or 44,000, respectively. The purified enzyme hydrolyzed PHB, P(3HB-co-3HV), and—at a very low rate—also PHV. Polyhydroxyalkanoates, PHA, with six or more carbon atoms per monomer or characteristic substrates for lipases were not hydrolyzed. In contrast to the PHB depolymerases ofPseudomonas lemoignei andAlcaligenes faecalis T₁, which are sensitive toward phenylmethylsulfonyl fluoride (PMSF) and which hydrolyze PHB mainly to the dimeric and trimeric esters of 3-hydroxybutyrate, the depolymerase ofComamonas sp. was insensitive toward PMSF and hydrolyzed PHB to monomeric 3-hydroxybutyrate indicating a different mechanism of PHB hydrolysis. Furthermore, the pH optimum of the reaction catalyzed by the depolymerase ofComamonas sp. was in the alkaline range at 9.4.