The effect of diphenylhydantoin upon degradation of sulphated macromolecules in cat palatal mucosa in vitro

The effect of diphenylhydantioin (DPH) upon the degradation of in vivo [35S]-sulphate-labelled proteoglycans was studied in cat palatal mucosa during organ culture. 8-week-old cats were injected intraperitoneally with [35S]-sulphate and 24 hours later the palatal mucosa was taken to organ culture. The release of radioactivity into the culture medium was taken as a measure of degradation of sulphated macromolecules, presumably proteoglycans, and the release of hydroxyproline as an indicator for collagen degradation. A parallel decrease in the release of radioactivity and in the release of hydroxyproline was observed when the culture was done in the presence of DPH (20 mg/l). Chromatography of the culture medium upon Sephadex G-25 revealed that the reduced release of radioactivity was due to a reduction of macromolecular degradation products leaving the amounts of free sulphate in the medium unchanged. The results were interpreted using a two compartment theory for proteoglycan degradation, extracellular breakdown of the protein core resulting in the production of macromolecular degradation products and intracellular lysosomal degradation resulting in free sulphate as the identifiable product. The results indicate that DPH inhibited the extracellular enzymatic degradation of proteoglycans without influencing their intracellular degradation.     

Biological degradation of 2,4-dichlorophenoxyacetic acid: chloride mass balance in stirred tank reactors.

A mass balance was developed for the degradation of 2,4-dichlorophenoxyacetic acid by a mixed culture. Batch culture experiments showed the degradation to be an acid-producing step. Inorganic chloride concentration consistently correlated with the expected value and with base consumption to maintain a constant pH.     

Influence of growth medium on cometabolic degradation of polycyclic aromatic hydrocarbons by <Emphasis Type="Italic">Sphingomonas</Emphasis> sp. strain PheB4

The influence of growth medium on cometabolic degradation of polycyclic aromatic hydrocarbons (PAHs) was investigated when Sphingomonas sp. strain PheB4 isolated from surface mangrove sediments was grown in either phenanthrene-containing mineral salts medium (PMSM) or nutrient broth (NB). The NB-grown culture exhibited a more rapid cometabolic degradation of single and mixed non-growth substrate PAHs compared to the PMSM-grown culture. The concentrations of PAH metabolites were also lower in NB-grown culture than in PMSM-grown culture, suggesting that NB-grown culture removed metabolites at a faster rate, particularly, for metabolites produced from cometabolic degradation of a binary mixture of PAHs. Cometabolic pathways of single PAH (anthracene, fluorene, or fluoranthene) in NB-grown culture showed similarity to that in PMSM-grown culture. However, cometabolic pathways of mixed PAHs were more diverse in NB-grown culture than that in PMSM-grown culture. These results indicated that nutrient rich medium was effective in enhancing cometabolic degradation of mixed PAHs concomitant with a rapid removal of metabolites, which could be useful for the bioremediation of mixed PAHs contaminated sites using Sphingomonas sp. strain PheB4.     

The role of benzoate in anaerobic degradation of terephthalate

The effects of acetate, benzoate, and periods without substrate on the anaerobic degradation of terephthalate (1, 4-benzene-dicarboxylate) by a syntrophic methanogenic culture were studied. The culture had been enriched on terephthalate and was capable of benzoate degradation without a lag phase. When incubated with a mixture of benzoate and terephthalate, subsequent degradation with preference for benzoate was observed. Both benzoate and acetate inhibited the anaerobic degradation of terephthalate. The observed inhibition is partially irreversible, resulting in a decrease (or even a complete loss) of the terephthalate-degrading activity after complete degradation of benzoate or acetate. Irreversible inhibition was characteristic for terephthalate degradation only because the inhibition of benzoate degradation by acetate could well be described by reversible noncompetitive product inhibition. Terephthalate degradation was furthermore irreversibly inhibited by periods without substrate of only a few hours. The inhibition of terephthalate degradation due to periods without substrate could be overcome through incubation of the culture with a mixture of benzoate and terephthalate. In this case no influence of a period without substrate was observed. Based on these observations it is postulated that decarboxylation of terephthalate, resulting in the formation of benzoate, is strictly dependent on the concomitant fermentation of benzoate. In the presence of higher concentrations of benzoate, however, benzoate is the favored substrate over terephthalate, and the culture loses its ability to degrade terephthalate. In order to overcome the inhibition of terephthalate degradation by benzoate and acetate, a two-stage reactor system is suggested for the treatment of wastewater generated during terephthalic acid production.     

Aerobic biodegradation of dinitrotoluenes in batch systems by pure and mixed cultures

Biodegradation characteristics of 2,4- and 2,6-dinitrotoluenes (DNTs) individually by pure strains and defined mixed cultures obtained from a mixed culture isolated from a slate packed bed bioreactor is described. Batch degradation experiments were carried out with free cells in submerged cultivations. The degradation rate and efficiency of five best individual bacterial strains, bacterial consortia comprising three and five of these strains, and the complete mixed culture were evaluated and compared. All the strains showed ability to degrade both the DNTs. All but one strain degraded both DNTs at the same rate. The degradation rate as well as the degradation efficiency by the mixed cultures was higher than that by the individual strains. The complete mixed culture showed 15-20x higher degradation rate than the individual bacterial strains.     

Effect of bioaugmentation and supplementary carbon sources on degradation of polycyclic aromatic hydrocarbons by a soil-derived culture

The degradation of polycyclic aromatic hydrocarbons (PAHs) by an undefined culture obtained from a PAH-polluted soil and the same culture bioaugmented with three PAH-degrading strains was studied in carbon-limited chemostat cultures. The PAHs were degraded efficiently by the soil culture and bioaugmentation did not significantly improve the PAH degrading performance. The presence of PAHs did, however, influence the bacterial composition of the bioaugmented and non-bioaugmented soil cultures, resulting in the increase in cell concentration of sphingomonad strains. the initial enhancement of the degradation of the PAHs by biostimulation gradually disappeared and only the presence of salicylate in the additional carbon sources had a lasting slightly stimulating effect on the degradation of phenanthrene. The results suggest that bioaugmentation and biostimulation have limited potential to enhance PAH bioremediation by culture already proficient in the degradation of such contaminants.     

Investigation of the biodegradation of pentachlorophenol by the predominant bacterial strains in a mixed culture

A pentachlorophenol (PCP) degrading mixed culture contained three predominant strains identified as Flavobacterium gleum, Agrobacterium radiobacter and Pseudomonas sp. The relative abilities of the three strains to degrade PCP were tested individually and in combination. Rates of PCP degradation by individual isolates were lower than that observed for the three isolates combined. Of the individual strains, Flavobacterium gleum manifested highest PCP degradation ability. A biodegradation medium inoculated with a combination of the three isolates exhibited PCP degradation patterns similar to the original mixed culture. Varying low amounts of tetrachlorophenol were found in degradation medium inoculated with individual isolates, but this intermediate was absent from media inoculated with the mixed culture.     

Perturbation of syntrophic isobutyrate and butyrate degradation with formate and hydrogen

The effect of formate and hydrogen on isomerization and syntrophic degradation of butyrate and isobutyrate was investigated using a defined methanogenic culture, consisting of syntrophic isobutyrate-butyrate degrader strain IB, Methanobacterium formicicum strain T1N, and Methanosarcina mazeii strain T18. Formate and hydrogen were used to perturb syntrophic butyrate and isobutyrate degradation by the culture. The reversible isomerization between isobutyrate and butyrate was inhibited by the addition of either formate or hydrogen, indicating that the isomerization was coupled with syntrophic butyrate degradation for the culture studied. Energetic analysis indicates that the direction of isomerization between isobutyrate and butyrate is controlled by the ratio between the two acids, and the most thermodynamically favorable condition for the degradation of butyrate or isobutyrate in conjunction with the isomerization is at almost equal concentrations of isobutyrate and butyrate. The degradation of isobutyrate and butyrate was completely inhibited in the presence of a high hydrogen partial pressure (>2000 Pa) or a measurable level of formate (10 muM or higher). Significant formate (more than 1 mM) was detected during the perturbation with hydrogen (17 to 40 kPa). Resumption of butyrate and isobutyrate degradation was related to the removal of formate. Energetic analysis supported that formate was another electron carrier, besides hydrogen, during syntrophic isobutyrate-butyrate degradation by this culture. (c) 1996 John Wiley & Sons, Inc.     

The pattern of collagen degradation in cultured tadpole tissues

A characteristic pattern of selective degradation of isotopically labeled collagen in tadpole tail fin in culture was observed by measuring the amount and radioactivity of degraded collagen fragments released into the culture medium as a function of time of incubation. The changes in specific activity and total amount of hydroxyproline released with time indicated early degradation apparent at 3 hr of incubation of a small fraction of newly synthesized heavily labeled collagen followed by breakdown of the bulk of old lightly labeled fibrils. Collagenase activity rose in the culture medium with the release of collagen breakdown products and continued long afterward. Serum in the medium significantly reduced the release of collagen degradation products to the medium and greatly lowered their specific activity. Possible mechanisms of selective collagen degradation are discussed.     

Enhanced anaerobic degradation of benzene by enrichment of mixed microbial culture and optimization of the culture medium

A heterogeneous mixed culture, originally collected from two different sources, namely cow-drug and sludge from the mineral medium containing 1% glucose and then adapted on benzene as the carbon and energy source. Under anaerobic conditions benzene was degraded via benzoic acid as a major intermediate in the benzene degradation pathway. The degradation rate of benzene was improved stepwise by the number of enrichments and optimization of the culture medium. The effects of microaerobic conditions and/or physicochemical treatment with H₂O₂ prior to anaerobic degradation were studied with respect to variations in benzene degradation rate, growth of biomass and gas produced is less than the theoretical value expected and the percentage of methane in the product gas was very small (3%–3.5%). The reason for this is not well understood but it is presumed that the major group of benzene-degrading bacteria present in the culture medium are sulphate reducers and the mixed consortium is unable to degrade certain complex aromatic intermediates in the benzene degradation pathway under the experimental conditions. For an actual explanation of the situation arising in this study, further investigations must be carrie out. However, the mixed culture is capable of oxidizing benzene more rapidly to intermediate compounds and also partly into gas under the culture conditions, compared to the published data for the anaerobic degradation of benzene.     

Dibutyl phthalate biodegradation by the white rot fungus, Polyporus brumalis

In this study, white rot fungus, Polyporus brumalis, was applied to degrade dibutyl phthalate (DBP), a major environmental pollutant. The degradation potential and resulting products were evaluated with HPLC and GC/MS. As DBP concentration increased to 250, 750, and 1,250 microM, the mycelial growth of P. brumalis was inhibited. However, growth was still observed in the 1,250 microM concentration. DBP was nearly eliminated from culture medium of P. brumalis within 12 days, with 50% of DBP adsorbed by the mycelium. Diethyl phthalate (DEP) and monobutyl phthalate (MBP) were detected as intermediate degradation products of DBP. In culture medium, the concentration of DEP was higher than that of MBP during the incubation period. After 12-15 days, the concentrations of both decreased rapidly in the culture medium. The primary final degradation product of DBP in culture medium was phthalic acid anhydride, as well as trace amounts of aromatic compounds, such as alpha-hydroxyphenylacetic acid, benzyl alcohol, and O-hydroxyphenylacetic acid. According to these results, the degradation of DBP in culture medium by the white rot fungus, P. brumalis, may be completed through two pathways-transesterification and de-esterification-which successively combine into an intracellular degradation pathway.     

Anaerobic degradation of 2-methylnaphthalene by a sulfate-reducing enrichment culture

Anaerobic degradation of 2-methylnaphthalene was investigated with a sulfate-reducing enrichment culture. Metabolite analyses revealed two groups of degradation products. The first group comprised two succinic acid adducts which were identified as naphthyl-2-methyl-succinic acid and naphthyl-2-methylene-succinic acid by comparison with chemically synthesized reference compounds. Naphthyl-2-methyl-succinic acid accumulated to 0.5 microM in culture supernatants. Production of naphthyl-2-methyl-succinic acid was analyzed in enzyme assays with dense cell suspensions. The conversion of 2-methylnaphthalene to naphthyl-2-methyl-succinic acid was detected at a specific activity of 0.020 +/- 0.003 nmol min(-1) mg of protein(-1) only in the presence of cells and fumarate. We conclude that under anaerobic conditions 2-methylnaphthalene is activated by fumarate addition to the methyl group, as is the case in anaerobic toluene degradation. The second group of metabolites comprised 2-naphthoic acid and reduced 2-naphthoic acid derivatives, including 5,6,7,8-tetrahydro-2-naphthoic acid, octahydro-2-naphthoic acid, and decahydro-2-naphthoic acid. These compounds were also identified in an earlier study as products of anaerobic naphthalene degradation with the same enrichment culture. A pathway for anaerobic degradation of 2-methylnaphthalene analogous to that for anaerobic toluene degradation is proposed.     

一株苯系物降解真菌筛选及降解特性

采用逐量分批驯化的方法以污水处理厂污泥作为菌源,苯、甲苯、二甲苯为唯一碳源,驯化、分离、筛选能够有效降解苯系物的真菌,命名为B1。采用单因素以及正交实验方法并对真菌降解环境影响因素及降解效率进行了测定和研究。结果表明:真菌B1对苯系物降解的最佳条件为C:N=5:1,pH5,温度30℃,菌种接种量为5.5ml(50ml培养基)。采用GC对初始液相浓度0~90mg/L范围内的苯系物降解效果进行测定,未发现苯系物对真菌降解活性产生抑制作用。真菌对苯系物的降解效率为:甲苯>苯>二甲苯,最高降解效率分别达到87.39%,85.21%,81.47%。混合物降解效果略高于单一底物的降解效果。     

The anaerobic biodegradation of diethanolamine by a nitrate reducing bacterium

The ability of bacterial cultures to degrade diethanolamine under anoxic conditions with nitrate as an electron acceptor was investigated. A mixed culture capable of anaerobic degradation of diethanolamine was obtained from river sediments by enrichment culture. From this a single bacterial strain was isolated which could use diethanolamine, monoethanolamine, triethanolamine and N-methyl diethanolamine as its sole carbon and energy sources either aerobically or anaerobically. Growth on diethanolamine was faster in the absence of oxygen. The accumulation of possible metabolites in the culture medium was determined as was the ability to grow on certain putative intermediates in the degradation of diethanolamine. A possible pathway for the degradation of ethanolamines by this organism is suggested.     

The anaerobic biodegradation of diethanolamine by a nitrate reducing bacterium

The ability of bacterial cultures to degrade diethanolamine under anoxic conditions with nitrate as an electron acceptor was investigated. A mixed culture capable of anaerobic degradation of diethanolamine was obtained from river sediments by enrichment culture. From this a single bacterial strain was isolated which could use diethanolamine, monoethanolamine, triethanolamine and N-methyl diethanolamine as its sole carbon and energy sources either aerobically or anaerobically. Growth on diethanolamine was faster in the absence of oxygen. The accumulation of possible metabolites in the culture medium was determined as was the ability to grow on certain putative intermediates in the degradation of diethanolamine. A possible pathway for the degradation of ethanolamines by this organism is suggested.     

Microbial metabolism of 2-chlorophenol, phenol and rho-cresol by Rhodococcus erythropolis M1 in co-culture with Pseudomonas fluorescens P1

Chlorophenolic waste most often contains phenol and rho-cresol along with chlorophenols. A Rhodococcus erythropolis strain M1 was isolated with the ability to degrade 2-chlorophenol, phenol and p-cresol (100 mgl(-1), each) in 18, 24 and 20 h, respectively, with negligible lag. However, Rhodococcus sp. characterized by low growth rate, pose a threat to be outgrown by bacteria occurring in natural habitats. In the present study, interaction of R. erythropolis M1 with another isolated bacteria generally encountered in activated sludge for water treatment like Pseudomonas fluorescens P1 was studied. 2-chlorophenol, phenol and p-cresol were selected as the substrates for the study. Viable cell counts showed competitive interaction between the species on 2-chlorophenol and phenol. Specific growth rate of pure culture of R. erythropolis M1 was higher than P. fluorescens P1 on 2-chlorophenol. However, in mixed culture, P. fluorescens P1 showed higher growth rate. Degradation of phenol showed higher growth rate of R. erythropolis M1 both in pure and in mixed culture form. Degradation of p-cresol had shown similar counts for both populations indicating neutral type of interaction. This observation was substantiated by detecting the growth rate, where both cultures had similar growth rate in pure and in the mixed culture form. Rate of 2-chlorophenol degradation was higher when R. erythropolis M1 was used as the pure culture as compared to the degradation rates observed with the P. fluorescens P1 or with the mixed culture. However, in case of phenol and p-cresol, degradation by the mixed culture had resulted in higher degradation rates as compared to the degradation of the substrates by both the axenic cultures.     

Growth kinetics of an indigenous mixed microbial consortium during phenol degradation in a batch reactor

Biodegradation of phenol by a mixed microbial culture, isolated from a sewage treatment plant, was investigated in batch shake flasks. A minimum concentration of 100 and a maximum of 800 mg 1(-1) of phenol in the media were adapted in the degradation study. The phenol degradation rate varied largely and was less than 10 mg l(-1)h(-1) at both extremes of the initial concentrations in the media. The degradation rate was maximum 15.7 mg l(-1)h(-1) at 400 mg l(-1) phenol. The culture followed substrate inhibition kinetics and the specific growth rate were fitted to Haldane and Han-Levenspiel models. Between the two models the Han-Levenspiel was found to be a better fit with a root mean square error of 0.0211. The biokinetics constants estimated using these models showed good potential of the mixed microbial culture in phenol degradation.     

Effect of pyocyanin on a crude-oil-degrading microbial community

Pseudomonas aeruginosa is an n-alkane degrader that is frequently isolated from petroleum-contaminated sites and produces factors that enhance its competitiveness and survival in many environments. In this study, one such factor, pyocyanin, has been detected in an oil-degrading culture containing P. aeruginosa and is a redox-active compound capable of inhibiting microbial growth. To examine the effects of pyocyanin further, an oil-degrading culture was grown with and without 9.5 microM pyocyanin and microbial community structure and oil degradation were monitored for 50 days. Denaturing gradient gel electrophoresis (DGGE) analysis of cultures revealed a decrease in the microbial community diversity in the pyocyanin-amended cultures compared to that of the unamended cultures. Two members of the microbial community in pure culture exhibited intermediate and high sensitivities to pyocyanin corresponding to intermediate and low levels of activity for the antioxidant enzymes catalase and superoxide dismutase, respectively. Another member of the community that remained constant in the DGGE gels over the 50-day culture incubation period exhibited no sensitivity to pyocyanin, corresponding to a high level of catalase and superoxide dismutase when examined in pure culture. Pyocyanin also affected the overall degradation of the crude oil. At 50 days, the culture without pyocyanin had decreased polycyclic aromatic hydrocarbons compared to the pyocyanin-amended culture, with a specific reduction in the degradation of dibenzothiophenes, naphthalenes, and C(29) and C(30) hopanes. This study demonstrated that pyocyanin influenced the diversity of the microbial community and suggests the importance of understanding how interspecies interactions influence the degradation capability of a microbial community.     

Continuous anaerobic phenol degradation using an adapted mixed culture

A mixed culture derived from cow dung and sewage sludge and adapted to phenol was used for anaerobic phenol degradation. The phenol degradation rate depended on the period of adaptation of the mixed culture to phenol. In the continuous process, a higher degradation rate (2500 mg.1^-1 d^-1) and better reactor stability was achieved with a granular activated-carbon-packed bed reactor than with a stirred tank reactor.The authors are with the Department of Biochemical Engineering & Biotechnology, Indian Institute of Technology, Delhi Hauz Khas, New Delhi, India.     

Anaerobic Degradation of 2-Methylnaphthalene by a Sulfate-Reducing Enrichment Culture

Anaerobic degradation of 2-methylnaphthalene was investigated with a sulfate-reducing enrichment culture. Metabolite analyses revealed two groups of degradation products. The first group comprised two succinic acid adducts which were identified as naphthyl-2-methyl-succinic acid and naphthyl-2-methylene-succinic acid by comparison with chemically synthesized reference compounds. Naphthyl-2-methyl-succinic acid accumulated to 0.5 μM in culture supernatants. Production of naphthyl-2-methyl-succinic acid was analyzed in enzyme assays with dense cell suspensions. The conversion of 2-methylnaphthalene to naphthyl-2-methyl-succinic acid was detected at a specific activity of 0.020 ± 0.003 nmol min⁻¹ mg of protein⁻¹ only in the presence of cells and fumarate. We conclude that under anaerobic conditions 2-methylnaphthalene is activated by fumarate addition to the methyl group, as is the case in anaerobic toluene degradation. The second group of metabolites comprised 2-naphthoic acid and reduced 2-naphthoic acid derivatives, including 5,6,7,8-tetrahydro-2-naphthoic acid, octahydro-2-naphthoic acid, and decahydro-2-naphthoic acid. These compounds were also identified in an earlier study as products of anaerobic naphthalene degradation with the same enrichment culture. A pathway for anaerobic degradation of 2-methylnaphthalene analogous to that for anaerobic toluene degradation is proposed.