Metabolism of mono- and dichloro-dibenzo-p-dioxins by Phanerochaete chrysosporium cytochromes P450

The white-rot fungus Phanerochaete chrysosporium possesses biodegradative capabilities of polychlorinated dibenzo-p-dioxins (PCDDs). One hundred twenty yeast clones expressing individual P450s of P. chrysosporum (PcCYPs), generated in our previous efforts, were screened for transformation of dioxin, and 40 positive clones were obtained. Of these clones, six clones showed metabolism of 2-chloro-dibenzo-p-dioxin, and a microsomal PcCYP designated as PcCYP11a3 showed much higher activity than any other PcCYPs. The turnover numbers of hydroxylation activities of PcCYP11a3 toward 1-MCDD (58 min⁻¹) and 2-MCDD (13 min⁻¹) are more than 200 times higher than those of previously reported PcCYP65a2. In addition, PcCYP11a3 catalyzes hydroxylation of 2,3-dichloro-dibenzo-p-dioxin. To our best knowledge, PcCYP11a3 has the highest activity toward PCDDs among the known CYPs derived from microorganisms. Although PcCYP11a3 showed no detectable activity toward 2,7-dichloro-dibenzo-p-dioxin and 2,3,7-trichloro-dibenzo-p-dioxin, PcCYP11a3 is promising as a template whose activity would be enhanced by site-directed mutagenesis.     

Removal of Dibenzofuran, Dibenzo-p-Dioxin, and 2-Chlorodibenzo-p-Dioxin from Soils Inoculated with Sphingomonas sp. Strain RW1

Removal of dibenzofuran, dibenzo-p-dioxin, and 2-chlorodibenzo-p-dioxin (2-CDD) (10 ppm each) from soil microcosms to final concentrations in the parts-per-billion range was affected by the addition of Sphingomonas sp. strain RW1. Rates and extents of removal were influenced by the density of RW1 organisms. For 2-CDD, the rate of removal was dependent on the content of soil organic matter (SOM), with half-life values ranging from 5.8 h (0% SOM) to 26.3 h (5.5% SOM).     

Biotransformation of dichloro-, trichloro-, and tetrachlorodibenzo-p-dioxin by the white-rot fungus Phlebia lindtneri

The model polychlorinated dibenzo-p-dioxins (PCDDs) 2,7-dichloro-, 2,3,7-trichloro, 1,2,6,7-, 1,2,8,9-, and 1,3,6,8-tetrachlorodibenzo-p-dioxin were used as substrates for a degradation experiment with the white-rot fungus Phlebia lindtneri. 2,7-Dichlorodibenzo-p-dioxin (2,7-diCDD) was biotransformed to hydroxylated diCDD and methoxylated diCDD. With the exception of 1,3,6,8-tetrachlorodibenzo-p-dioxin, the tri- and tetrachlorodibenzo-p-dioxins were biotransformed to hydroxyl and methoxyl compounds by P. lindtneri. The degradation rate of 1,2,6,7-tetrachlorodibenzo-p-dioxin was higher than that of 2,3,7-trichlorodibenzo-p-dioxin and no degradation of 1,3,6,8-tetrachlorodibenzo-p-dioxin was observed. These results indicate that the degradation of these PCDDs depends on the chlorination patterns of the substrates. This is the first report of the hydroxylation and methoxylation of tri- to tetra-CDDs by a fungal strain.     

Hydrolysis of β-1,3/1,6-glucan by glycoside hydrolase family 16 endo-1,3(4)-β-glucanase from the basidiomycete Phanerochaete chrysosporium

When Phanerochaete chrysosporium was grown with laminarin (a β-1,3/1,6-glucan) as the sole carbon source, a β-1,3-glucanase with a molecular mass of 36 kDa was produced as a major extracellular protein. The cDNA encoding this enzyme was cloned, and the deduced amino acid sequence revealed that this enzyme belongs to glycoside hydrolase family 16; it was named Lam16A. Recombinant Lam16A, expressed in the methylotrophic yeast Pichia pastoris, randomly hydrolyzes linear β-1,3-glucan, branched β-1,3/1,6-glucan, and β-1,3-1,4-glucan, suggesting that the enzyme is a typical endo-1,3(4)-β-glucanase (EC 3.2.1.6) with broad substrate specificity for β-1,3-glucans. When laminarin and lichenan were used as substrates, Lam16A produced 6-O-glucosyl-laminaritriose (β-d-Glcp-(1–>6)-β-d-Glcp-(1–>3)-β-d-Glcp-(1–>3)-d-Glc) and 4-O-glucosyl-laminaribiose (β-d-Glcp-(1–>4)-β-d-Glcp-(1–>3)-d-Glc), respectively, as one of the major products. These results suggested that the enzyme strictly recognizes β-d-Glcp-(1–>3)-d-Glcp at subsites −2 and −1, whereas it permits 6-O-glucosyl substitution at subsite +1 and a β-1,4-glucosidic linkage at the catalytic site. Consequently, Lam16A generates non-branched oligosaccharide from branched β-1,3/1,6-glucan and, thus, may contribute to the effective degradation of such molecules in combination with other extracellular β-1,3-glucanases.     

The microbial degradation of chlorophenolic preservatives in spent,pressure-treated timber

The reduction of pentachlorophenol in treated timber, after inoculation with pentachlorophenol-degrading bacterial species,Rhodococcus chlorophenolicus andFlavobacterium sp., and the white-rot fungusPhanerochaete chrysosporium, was monitored in solid substrate systems and in liquid culture suspensions. In solid substrate systems there was no significant pentachlorophenol degradation by the bacterial species under a variety of conditions. Under similar conditions,Phanerochaete chrysosporium transformed over 80% of the starting concentration of 500 ppm to pentachloroanisole. In liquid culture suspensions however, mid-exponential phaseFlavobacterium sp. cells were able to degrade over 99% of the pentachlorophenol in sawdust and wood chips due to the extraction of PCP from the timber as a water soluble salt. There were however no significant changes in the chlorinated dioxin components during this treatment.Abbreviations ATTC American type culture collection - AWPA American Wood Preservers' Association - DSM Deutsche Sammlung für Mikroorganismen - GC/MS gas chromatograph/mass spectrometer - HpCDD heptachlorodibenzo-p-dioxin - HpCDF heptachlorodibenzofuran - HxCDD hexachlorodibenzo-p-dioxin - HxCDF hexachlorodibenzofuran - ¹³C-OCDD carbon 13-labelled octachlorodibenzo-p-dioxin - OCDD octachlorodibenzo-p-dioxin - OCDF octachlorodibenzofuran - PCDDs polychlorinated dibenzo-p-dioxins - PCDFs polychlorinated dibenzofurans - PCP pentachlorophenol - PnCDD pentachlorodibenzo-p-dioxin - PnCDF pentachlorodibenzofuran - TCDD tetrachlorodibenzo-p-dioxin - TCDF terachlorodibenzofuran - TeCP tetrachlorophenol - WHC water holding capacity - w/v weight for volume ratio     

Superior survival and degradation of dibenzo-p-dioxin and dibenzofuran in soil by soil-adapted Sphingomonas sp. strain RW1

The dibenzo-p-dioxin(DD)- and dibenzofuran(DF)-degrading bacterium, Sphingomonas sp. strain RW1, was tagged by insertion of a mini-Tn5 lacZ transposon in order to follow its fate in complex laboratory soil systems. The tagged strain was tested for its ability to survive in soil and degrade DF and DD applied at a concentration of 1 mg/g. Bacteria pre-adapted to soil conditions were found to survive better in DF- and DD-amended soil and degrade the substrate more efficiently than bacteria that had not been subjected to pre-adaptation. The concentration of soil-applied DF and DD, individually and in combination, decreased to less than 2% of the original concentrations within 3 weeks of addition of the RW1 derivative, accompanied by a short, but significant exponential increase in RW1 viable cells. During the same period the native bacterial population in soil was stable while viable fungi declined. Received: 12 November 1996 / Received revision: 21 February 1997 / Accepted: 22 February 1997     

CYP1B1 (4326C > G), CYP2F1 (c.14_15insC), CYP2J2 (?76G > T), and CYP2S1 (13106C > T and 13255A > G) polymorphisms and genetic predisposition to chronic respiratory diseases induced by smoking and occupational factors

The contribution of the polymorphic markers of cytochrome P450 genes to respiratory diseases caused by smoking and occupational factors has been assessed. For this purpose, PCR-RFLP analysis of the CYP1B1 (rs1056836, 4326C > G), CYP2F1 (rs11399890, c.14_15insC), CYP2J2 (rs890293, -76G > T), and CYP2S1 (rs34971233, 13106C > T and rs338583, 13255A > G) gene polymorphisms has been performed. The analysis has shown that CYP1B1 (rs1056836, 4326C > G) and CYP2F1 (rs11399890, c.14_15insC) polymorphisms may contribute to the development of occupational chronic bronchitis. The proportion of CYP1B1*1*3 heterozygotes in the group of patients with occupational chronic bronchitis is considerably greater than in the group of healthy workers (69.16% versus 53.29%; χ² = 5.94, p = 0.02, p _cur = 0.04, OR = 1.97, the 95% CI is 1.13–3.42). Patients with occupational chronic bronchitis and healthy workers significantly differed from each other in the CYP2F1 genotypes frequency distribution (rs11399890, c.14_15insC) (χ² = 6.18, d.f. = 2, p = 0.05). CYP2F1 wild type/ins heterozygous genotype frequency is higher in healthy workers (36.08%) than in patients (22.22%) (χ² = 5.48, p = 0.02, p _cur = 0.04, OR = 0.51, the 95% CI is 0.28–0.90). No association has been found between the CYP2J2 (rs890293, −76G > T) or CYP2S1 (rs34971233, 13106C > T, and rs338583, 13255A > G) gene polymorphisms and respiratory diseases.     

鼠李糖乳杆菌碱性磷酸酶的提取条件优化及其降解有机磷农药作用

【背景】我国作为农业大国,对农药的大量使用是不可避免的,但是农药的超范围使用、超标及高检出率对于环境的污染与人体健康的威胁日趋严重。【目的】碱性磷酸酶(alkaline phosphatase,ALP)对有机磷农药具有积极的降解作用,因此,本文对鼠李糖乳杆菌(Lactobacillus rhamnosus) Z23(LGG Z23)所产碱性磷酸酶的提取条件进行优化,并研究其对有机磷农药的降解作用。【方法】使用单因素试验和正交试验优化ALP的提取条件;使用对硝基苯酚法测定酶活力;使用分级沉淀和层析法提纯ALP;使用乙酰胆碱酯酶抑制法测定ALP对有机磷农药的降解率。【结果】LGG Z23所产ALP的最优提取条件为：细胞破碎时间15 min,破碎功率450 W,料液比(质量体积比)1:6,提取液pH 10.0,此条件下酶活力为(4.95±0.26) U/mL,比优化前提高2.11倍;对6种有机磷农药的降解率效果为敌敌畏(95.79%±0.01%)>甲基对硫磷(90.69%±0.03%)>毒死蜱(88.90%±0.02%)>敌百虫(86.07%±0.03%)>马拉硫磷(85.31%±0.02%)>乐果(83.18%±0.03%),其中对敌敌畏和甲基对硫磷的降解效果最好,可达90%以上,并且降解作用差异显著(P<0.05)。【结论】本研究为LGG Z23所产ALP的应用研究提供了理论依据和实验数据。     

Mesophilic and thermophilic BTEX substrate interactions for a toluene-acclimatized biofilter

Benzene, toluene, ethylbenzene and xylene (BTEX) substrate interactions for a mesophilic (25°C) and thermophilic (50°C) toluene-acclimatized composted pine bark biofilter were investigated. Toluene, benzene, ethylbenzene, o-xylene, m-xylene and p-xylene removal efficiencies, both individually and in paired mixtures with toluene (1:1 ratio), were determined at a total loading rate of 18.1 g m^–3 h^–1 and retention time ranges of 0.5–3.0 min and 0.6–3.8 min for mesophilic and thermophilic biofilters, respectively. Overall, toluene degradation rates under mesophilic conditions were superior to degradation rates of individual BEX compounds. With the exception of p-xylene, higher removal efficiencies were achieved for individual BEX compounds compared to toluene under thermophilic conditions. Overall BEX compound degradation under mesophilic conditions was ranked as ethylbenzene >benzene >o-xylene >m-xylene >p-xylene. Under thermophilic conditions overall BEX compound degradation was ranked as benzene >o-xylene >ethylbenzene >m-xylene >p-xylene. With the exception of o-xylene, the presence of toluene in paired mixtures with BEX compounds resulted in enhanced removal efficiencies of BEX compounds, under both mesophilic and thermophilic conditions. A substrate interaction index was calculated to compare removal efficiencies at a retention time of 0.8 min (50 s). A reduction in toluene removal efficiencies (negative interaction) in the presence of individual BEX compounds was observed under mesophilic conditions, while enhanced toluene removal efficiency was achieved in the presence of other BEX compounds, with the exception of p-xylene under thermophilic conditions.     

Biotransformation of naphthalene and diaryl ethers by green microalgae

The role of green microalgae in thebiotransformation of naphthalene (a polycyclicaromatic hydrocarbon) and diaryl ethers wasinvestigated using axenic cultures ofChlorella vulgaris and two environmentalisolates, Scenedesmus SI1 andAnkistrodesmus SI2.Biotransformation experiments with dense cell culturesshowed that these three greenalgae transformed toxic xenobiotics to more polar metabolites.Chlorella vulgarismetabolized naphthalene to 1-naphthol (0.36–0.65%). Ankistrodesmus SI2 biotransformed dibenzofuran tosix metabolites (total over7%), three of which (possibly four) were identified as monohydroxylated dibenzofurans, the remaining two may be dihydroxylated derivatives. Scenedesmus SI1 biotransformeddibenzo-p-dioxin to three metabolites, one ofwhich was tentatively identified as2-hydroxydibenzo-p-dioxin (approximately 3.8%),the remainder may be dihydroxylated derivatives.This is the first time that the biotransformation of diaryl ethers by green microalgae has been investigated.     

Characterization of an acid-labile, thermostable β-glycosidase from Thermoplasma acidophilum

A recombinant putative β-galactosidase from Thermoplasma acidophilum was purified as a single 57 kDa band of 82 U mg⁻¹. The molecular mass of the native enzyme was 114 kDa as a dimer. Maximum activity was observed at pH 6.0 and 90°C. The enzyme was unstable below pH 6.0: at pH 6 its half-life at 75°C was 28 days but at pH 4.5 was only 13 h. Catalytic efficiencies decreased as p-nitrophenyl(pNP)-β-d-fucopyranoside (1067) > pNP-β-d-glucopyranoside (381) > pNP-β-d-galactopyranoside (18) > pNP-β-d-mannopyranoside (11 s⁻¹ mM⁻¹), indicating that the enzyme was a β-glycosidase.     

Biochemical and Clinical Improvement of Cytotoxic State by Amantadine Sulphate

  1. The main idea of the open clinical trial was to compare the income and outcome clinical picture and the evolution of the biochemical markers in the defined intervals in closed head injury group patients.2. In the group of 32 patients, mean age 40.78±15.36 years suffering from closed traumatic brain injury the following markers were measured: glycaemia, malondialdehyde (MDA) as marker of lipid peroxidation, beta-caroten, total SH groups as marker of protein oxidation in the following intervals: between the 1st and the 3rd, between the 3rd and the 7th, between the 1st and the 7th day respectively.3. Glycaemia significantly decreased since the 1st day till the 3rd day (p < 0.05) and since the 1st day till the 7th day (p < 0.05) but it was not significantly changed since the 3rd day till the 7th day (p > 0.05).4. MDA 1st × MDA 3rd p > 0.05 insignificant change, MDA 1st × MDA 7th p < 0.001—high significant decrease, MDA 3rd × MDA 7th—p < 0.0001—very high significant decrease.5. Beta-caroten the 1st × beta-caroten the 3rd day was insignificantly changed—p > 0.05, the 3rd × the 7th day beta-caroten increased significantly—p < 0.0002, the 1st day × 7th day beta-caroten significantly increased—p < 0.0001.6. We examined the SH groups only in nine patients, due to technical problems and SH groups decrease on the 3rd day (p < 0.005).7. In 18 amantadine sulphate subgroups (randomly selected), there was 5.5% lethality and mean outcome GCS (outGCS) 9.83±3.8, while lethality of the control subgroup (n=14) was 42.9%, mean outGCS 6.28±3.5.     

Biodegradation of dioxins by recombinant Escherichia coli expressing rat CYP1A1 or its mutant

Among polychlorinated dibenzo-p-dioxins (PCDDs), 2,3,7,8-tetrachlorodibenzo-p-dioxin (2,3,7,8-TetraCDD) is the most toxic one. Recently, we reported that rat CYP1A1 mutant, F240A, expressed in yeast showed metabolic activity toward 2,3,7,8-TetraCDD. In this study, we successfully expressed N-terminal truncated P450s (Δ1A1 and ΔF240A) in Escherichia coli cells. Kinetic analysis using membrane fractions prepared from the recombinant E. coli cells revealed that ΔF240A has enzymatic properties similar to F240A expressed in yeast. The metabolism of PCDDs by recombinant E. coli cells expressing both ΔF240A and human NADPH-P450 reductase was also examined. When 2,3,7-TriCDD was added to the E. coli cell culture at a final concentration of 10 μM, approximately 90% of the 2,3,7-TriCDD was converted into multiple metabolites within 8 h. These results indicate the possible application of prokaryotic cells expressing ΔF240A to the bioremediation of PCDD-contaminated soil.     

The effects of selenium deficiency on differentiation, degradation, dand cell lysis of L8 rat skeletal muscle cells

We investigated the effects of selenium (Se) deficiency on differentiation, protein degradation, and cell lysis in cultured skeletal muscle cells, using L8 rat skeletal muscle cells cultured in serum-free (SF) medium to induce differentiation and to maintain myotubes. Creatine kinase activity was reduced (p < 0.05) by approximately 15% without Se supplementation for 96 h. Confluent myoblasts were treated with SF media with four different levels of vitamin E (0,10, 35, and 100 μM) in the absence and presence of Se (0 and 0.25 μM, respectively). After 96 h, vitamin E at a high dose (100 μM) was effective in the prevention of the decrease of differentiation caused by Se deficiency (p < 0.05). Following differentiation, the effects of three Se concentrations (0, 0.25, and 2.5 μM) on degradation of proteins as assessed by release of³H-labeled free amino acids secreted into the media were studied. Selenium supplementation did not affect (p > 0.05) total protein degradation. However, Se deficiency increased (p < 0.05) lactate dehydrogenase released from lyzed dead cells. The results indicate that Se is required to maintain an optimal rate of muscle cell differentiation and health of myotube cultures.     

Characterization of a strain of <Emphasis Type="Italic">Pseudomonas putida</Emphasis> isolated from agricultural soil that degrades cadusafos (an organophosphorus pesticide)

Bacteria capable of degrading the pesticide, cadusafos, were isolated from agricultural soil using an enrichment method. In this way, five distinct cadusafos-degrading strains of Pseudomonas putidia were isolated, and were characterized using morphological and biochemical analysis, as well as 16S rRNA sequencing. Strain PC1 exhibited the greatest cadusafos degradation rate and was consequently selected for further investigation. Degradation of cadusafos by strain PC1 was rapid at 20 and 37°C, but was greatly reduced (~1.5-fold) by the presence of carbon sources. Strain PC1 was able to effectively degrade cadusafos in sterilized soil using low inoculum levels. The maximum degradation rate of cadusafos (V _max ) was calculated as 1.1 mg l⁻¹ day⁻¹, and its saturation constant (K _s ) was determined as 2.5 mg l⁻¹. Bacteria such as strain PC1, that use cadusafos as a carbon source, could be employed for the bioremediation of sites contaminated with pesticides.     

Isolation of a Xanthobacter sp. degrading dichloromethane and characterization of the gene involved in the degradation

A bacterial strain able to degrade dichloromethane (DCM) as the sole carbon source was isolated from a wastewater treatment plant receiving domestic and pharmaceutical effluent. 16S rDNA studies revealed the strain to be a Xanthobacter sp. (strain TM1). The new isolated strain when grown aerobically on DCM showed Luong type growth kinetics, with μ_max of 0.094 h⁻¹ and S _m of 1,435 mg l⁻¹. Strain TM1 was able to degrade other aromatic and aliphatic halogenated compounds, such as halobenzoates, 2-chloroethanol and dichloroethane. The gene for DCM dehalogenase, which is the key enzyme in DCM degradation, was amplified through PCR reactions. Strain TM1 contains type A DCM dehalogenase (dcmAa), while no product could be obtained for type B dehalogense (dcmAb). The sequence was compared against 12 dcmAa from other DCM degrading strains and 98% or 99% similarity was observed with all other previously isolated DCM dehalogenase genes. This is the first time a Xanthobacter sp. is reported to degrade DCM.     

Isolation of a buprofezin co-metabolizing strain of <Emphasis Type="Italic">Pseudomonas</Emphasis> sp. DFS35-4 and identification of the buprofezin transformation pathway

Buprofezin is a widely used insecticide that has caused environmental pollution in many areas. However, biodegradation of buprofezin by pure cultures has not been extensively studied, and the transformation pathway of buprofezin remains unclear. In this paper, a buprofezin co-metabolizing strain of DFS35-4 was isolated from a buprofezin-polluted soil in China. Strain DFS35-4 was preliminarily identified as Pseudomonas sp. based on its morphological, physiological, and biochemical properties, as well as 16S rRNA gene analysis. In the presence of 2.0 g l⁻¹ sodium citrate, strain DFS35-4 degraded over 70% of 50 mg l⁻¹ buprofezin in 3 days. Strain DFS35-4 efficiently degraded buprofezin in the pH range of 5.0–10.0 and at temperatures between 20 and 30°C. Three metabolites, 2-imino-5-phenyl-3-(propan-2-yl)-1,3,5-thiadiazinan-4-one, 2-imino-5-phenyl-1,3,5-thiadiazinan-4-one, and methyl(phenyl) carbamic acid, were identified during the degradation of buprofezin using gas chromatography–mass spectrometry (GC–MS) and tandem mass spectrometry (MS/MS). A partial transformation pathway of buprofezin in Pseudomonas sp. DFS35-4 was proposed based on these metabolites.     

Metabolism of polychlorinated dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxins by cytochrome P450 BM-3 and its mutant

The metabolism of polychlorinated dibenzo-p-dioxins by cytochrome P450 BM-3 from Bacillus megaterium and a mutant enzyme of it (AL₄V; Ala74Gly, Phe87Val, Leu188Gln triple mutant) was examined. Both purified enzymes metabolized 1-monochloro-, 2,3-dichloro-, and 2,3,7-trichloro-dibenzo-p-dioxin, but not 2,3,7,8-tetrachloro-dibenzo-p-dioxin. The mutant AL ₄V had 2–12 times higher activity than the wild-type P450 BM-3 towards polychlorinated dibenzo-p-dioxins. The products were hydroxylated at an unsubstituted position and/or showing migration of the chloride and were less toxic derivatives with lower than 10% toxicity of the original compounds.Revisions requested 26 August 2004; Revisions received 15 October 2004     

Benzene/toluene/p-xylene degradation. Part II. Effect of substrate interactions and feeding strategies in toluene/benzene and toluene/p-xylene fermentations in a partitioning bioreactor

A two-phase aqueous/organic partitioning bioreactor scheme was used to degrade mixtures of toluene and benzene, and toluene and p-xylene, using simultaneous and sequential feeding strategies. The aqueous phase of the partitioning bioreactor contained Pseudomonas sp. ATCC 55595, an organism able to degrade benzene, toluene and p-xylene simultaneously. An industrial grade of oleyl alcohol served as the organic phase. In each experiment, the organic phase of the bioreactor was loaded with 10.15 g toluene, and either 2.0 g benzene or 2.1 g p-xylene. The resulting aqueous phase concentrations were 50 mg/l, 25 mg/l and 8 mg/l toluene, benzene and p-xylene respectively. The simultaneous fermentation of benzene and toluene consumed these compounds at volumetric rates of 0.024 g l⁻¹ h⁻¹ and 0.067 g l⁻¹ h⁻¹, respectively. The simultaneous fermentation of toluene and p-xylene consumed these xenobiotics at volumetric rates of 0.066 g l⁻¹ h⁻¹ and 0.018 g l⁻¹ h⁻¹, respectively. A sequential feeding strategy was employed in which toluene was added initially, but the benzene or p-xylene aliquot was added only after the cells had consumed half of the initial toluene concentration. This strategy was shown to improve overall degradation rates, and to reduce the stress on the microorganisms. In the sequential fermentation of benzene and toluene, the volumetric degradation rates were 0.056 g l⁻¹ h⁻¹ and 0.079 g l⁻¹ h⁻¹, respectively. In the toluene/p-xylene sequential fermentation, the initial toluene load was consumed before the p-xylene aliquot was consumed. After 12 h in which no p-xylene degradation was observed, a 4.0-g toluene aliquot was added, and p-xylene degradation resumed. Excluding that 12-h period, the microbes consumed toluene and p-xylene at volumetric rates of 0.074 g l⁻¹ h⁻¹ and 0.025 g l⁻¹ h⁻¹, respectively. Oxygen limitation occurred in all fermentations during the rapid growth phase. Received: 16 November 1998 / Received revision: 29 March 1999 / Accepted: 9 April 1999     

Biotransformation of polychlorinated dibenzo-<Emphasis Type="Italic">p</Emphasis>-dioxin by <Emphasis Type="Italic">Coprinellus</Emphasis> species

A degradation experiment on dibenzo-p-dioxin (DD) and 2,7-dichlorodibenzo-p-dioxin (2,7-DCDD) was carried out using basidiomycetous fungi belonging to the genera Coprinus, Coprinellus, and Coprinopsis. Some species showed a high rate of decrease in DD for the 2-week test period. Among them, Coprinellus disseminatus showed the highest ability to decrease the DD level, close to 100% by the end of 2 weeks. Further examination showed that Coprinellus disseminatus and Coprinellus micaceus, belonging to the genus Coprinellus, were able to metabolize 2,7-DCDD to a monohydroxylated compound, probably mediated by the P450 system. The metabolism of chlorinated DD by fungi capable of living in soil conditions is reported here for the first time.