RESUMO
Heterogeneous composite wastes from landfills were evaluated as precursors for the generation of activated carbon (AC). A single-step chemical activation process was applied involving irradiation with microwave energy and impregnation with KOH. The average percentage yield of AC from active landfill precursor was higher than that from closed landfill for all depths sampled. Increase in impregnation ratio and irradiation power decreased the average percentage yield for both landfill precursors (active: 38.1 to 33.1%; closed: 42.1 to 33.3%). The optimum pH range for adsorption of methylene blue was pH 6-7, while adsorption increased with increase in temperature over the range 30 to 50°C. Carbonyl and hydroxyl groups were the major functional groups on the surface of AC. The properties of the AC are potentially suitable for the removal of cationic dyes and pollutants. AC generated from the landfill composite was comparable to that from other biomass being managed through AC generation. This is the first report to demonstrate the possible reuse of landfill composite as AC. The reuse option of landfill composite could provide a means of sustainable management of landfilled municipal waste.
Assuntos
Resíduos Sólidos , Instalações de Eliminação de Resíduos , Adsorção , Carbono , Carvão Vegetal , TemperaturaRESUMO
AIM: To evaluate possible source of nutrients for bacterial growth within polyurethane (PU) foam of used cot mattresses as determinants of bacterial population density. METHODS AND RESULTS: Used infant mattresses (n = 30) were analysed for bacteria capable of degrading colloidal PU and for aqueous soluble chemical components (aromatic amines, ammonium ions, phosphates and protein). Mattress type (waterproof cover vs exposed PU foam at the infant-head region), mattress age and previous use by another child were evaluated as factors that could influence the measured parameters. The levels of protein extracted from PU foam were (i) significantly (P = 0.0019) higher for mattresses lacking a waterproof cover at the infant-head region and (ii) positively correlated with both culturable bacterial population densities of the PU foams and extent of growth of Staphylococcus aureus on aqueous leachates. No statistically significant (P > 0.05) associations between other measured parameters and mattress type/use factors were identified. CONCLUSIONS: Infant use of cot mattresses with exposed PU foam leads to accumulation of proteins within the PU, which can promote bacterial growth. SIGNIFICANCE AND IMPACT OF THE STUDY: The study provides a mechanistic explanation for increased levels of bacteria associated with exposed PU of cot mattresses. In the context of the common bacterial toxins hypothesis for the sudden infant death syndrome (SIDS), this could explain the lowered risk of SIDS associated with use of a waterproof cover above the mattress.
Assuntos
Infecções Bacterianas/transmissão , Leitos/microbiologia , Equipamentos para Lactente/microbiologia , Poliuretanos , Proteínas/metabolismo , Staphylococcus aureus/metabolismo , Bactérias/isolamento & purificação , Bactérias/metabolismo , Pré-Escolar , Contagem de Colônia Microbiana , Reservatórios de Doenças , Contaminação de Equipamentos , Fungos/fisiologia , Humanos , Umidade , Lactente , Recém-Nascido , Viabilidade Microbiana , Staphylococcus aureus/isolamento & purificação , Morte Súbita do Lactente/etiologiaRESUMO
Multivariate analysis of a heavy metal pollution survey of closed and active landfill precursors was carried out in order to compare environmental risk levels in relation to age, particle size and depth of the precursors. Landfill precursors (77) were collected and analyzed for 15 USEPA toxic heavy metals using ICP-MS. Heavy metals concentrations in closed landfill precursors were significantly higher than those in the active landfill for 11 of 15 heavy metals investigated (closed landfill order: Feâ¯>â¯Alâ¯>â¯Mnâ¯>â¯Cuâ¯>â¯Pbâ¯>â¯Baâ¯>â¯Coâ¯>â¯Crâ¯>â¯Niâ¯>â¯Cdâ¯>â¯Asâ¯>â¯Seâ¯>â¯Ti). Cluster analysis and correlation studies indicated the distribution of the metals was more influenced by landfill precursor size than by depth of the sample. Principal component analysis (PCA) showed that 10 of 15 of heavy metals of both landfill precursors were from similar anthropogenic sources. Heavy metals pollution indices (Igeoâ¯>â¯5, EFâ¯>â¯40 and CFâ¯>â¯7) of both active and closed landfill precursors exceeded limits in the order of Znâ¯>â¯Cdâ¯>â¯Pbâ¯>â¯Cuâ¯>â¯Ag, indicating a major potential health risk influenced by age and particle size of precursor. Zn, Cd, Cu and Pb of both landfill precursors exceeded the USEPA set standard for assessment of human health risk for each of the metals (1â¯×â¯10-4 to 1â¯×â¯10-3). This study highlights the need for the integration of a clean-up process for precursors from both types of landfill to reduce possible environmental pollution during a reuse process.
RESUMO
Mitochondrial aconitase has been shown to be inactivated by a spectrum of substances or critical states. Fluoroacetate (FA) is the most known toxic agent inhibiting aconitase. The biochemistry of toxic action of FA is rather well understood, though no effective therapy has been proposed for the past six decades. In order to reveal novel approaches for possible antidotes to be developed, experiments were performed with rat liver mitochondria, Ehrlich ascite tumor cells and cardiomyocytes, exposed to FA or fluorocitrate in vitro. The effect of FA developed at much higher concentrations in comparison with fluorocitrate and was dependent upon respiratory substrates in experiments with mitochondria: with pyruvate, FA induced a slow oxidation and/or leak of pyridine nucleotides and inhibition of respiration. Oxidation of pyridine nucleotides was prevented by incubation of mitochondria with cyclosporin A. Studies of the pyridine nucleotides level and calcium response generated in Ehrlich ascite tumor cells under activation with ATP also revealed a loss of pyridine nucleotides from mitochondria resulting in a shift in the balance of mitochondrial and cytosolic NAD(P)H under exposure to FA. An increase of cytosolic [Ca2+] was observed in the cell lines exposed to FA and is explained by activation of plasma membrane calcium channels; this mechanism, could have an impact on amplitude and rate of Ca2+ waves in cardiomyocytes. Highlighting the reciprocal relationship between intracellular pyridine nucleotides and calcium balance, we discuss metabolic pathway modulation in the context of probable development of an effective therapy for FA poisoning and other inhibitors of aconitase.
Assuntos
Aconitato Hidratase/antagonistas & inibidores , Aconitato Hidratase/efeitos dos fármacos , Fluoracetatos/farmacologia , Mitocôndrias Hepáticas/enzimologia , Animais , Cálcio/metabolismo , Carcinoma de Ehrlich/metabolismo , Potenciais da Membrana/efeitos dos fármacos , Redes e Vias Metabólicas/efeitos dos fármacos , Mitocôndrias Hepáticas/efeitos dos fármacos , Mitocôndrias Hepáticas/metabolismo , Miócitos Cardíacos/efeitos dos fármacos , Miócitos Cardíacos/metabolismo , NADP/metabolismo , Oxirredução/efeitos dos fármacos , Ratos , Ratos WistarRESUMO
A microbial basis for bioreductive generation of phosphine is proposed, which could account at least in part for the presence of this toxic gas in natural anaerobic environments and in sewage and landfill gases. Phosphine generation under anaerobic growth conditions was dependent upon both the culture inoculum source (animal faeces) and enrichment culture conditions. Phosphine was detected in headspace gases from mixed cultures under conditions promoting fermentative growth of mixed acid and butyric acid bacteria, either in the presence or absence of methane generation. Monoseptic cultures of certain mixed acid fermentors (Escherichia coli, Salmonella gallinarum, and Salmonella arizonae) and solvent fermentors (Clostridium sporogenes, Clostridium acetobutyricum and Clostridium cochliarium) also generated phosphine. Such fermentative bacteria participate in the multi-stage process of methanogenesis in nature. Generation of phosphine by these bacteria, rather than by methanoarchaea themselves, could explain the apparent correlation between methanogenesis and the formation of phosphine in nature.
Assuntos
Bactérias Anaeróbias/metabolismo , Fosfinas/metabolismo , Esgotos , Poluentes Ambientais/análise , Fermentação , Oxirredução , Microbiologia do SoloRESUMO
Trimethylantimony was detected by gas chromatography-mass spectrometry (GC-MS) in the headspace of a soil enrichment culture designed to promote growth of clostridia. Clostridial isolates from the soil enrichment culture were shown to biomethylate inorganic antimony in monseptic culture, using hydride generation-gas chromatographyatomic absorption spectrometry (HG-GC-AAS). GC-MS profiles of headspace gases from soil enrichment cultures shown to generate trimethylantimony, were used to select characterised Clostridium spp for assessment of antimony biomethylation capability. Involatile methylantimony species (up to 21 microg Sb dm(-3)) were detected by HG-GC-AAS in the medium of monoseptic cultures of C. acetobutylicum, C. butyricum and C. cochlearium. The relative quantities of involatile mono-, di- and trimethylantimony species produced over the course of a 28-day cultivation period is consistent with trimethylantimony oxide being a final product of antimony biomethylation by these bacteria, with mono- and dimethylantimony species appearing transiently in the cultures as intermediates of an antimony biomethylation pathway. Clostridia may be the principal agents of antimony biomethylation in methanogenic environments and could give rise to methylated forms of antimony in both the aqueous and gaseous phases.
Assuntos
Clostridium/fisiologia , Compostos Organometálicos/química , Poluentes do Solo/análise , Disponibilidade Biológica , Cromatografia Gasosa-Espectrometria de Massas , Metilação , Microbiologia do SoloRESUMO
1. The aerobic filamentous fungus S. brevicaulis IMI 17297 methylated antimony from Sb2O3 substrate, with the formation of gaseous trimethylantimony (TMA). No evidence was found for the generation of other gaseous antimony compounds by this organism. 2. Biovolatilization of inorganic antimony was greatest during cultivation of the fungus on solid media at 25 degrees C, and occurred more readily from antimony (III) substrates than from antimony (V) substrates. 3. Under simulated cot environment conditions (CO2 enriched atmosphere, 33 degrees C) the fungus exhibited an altered morphology and a reduced capability to volatilize inorganic antimony from the pure compound. 4. No evidence of antimony biovolatilization from cot mattress PVC was found, unless antimony was released from PVC by heat treatment (at 80 or 100 degrees C). 5. These data suggest that normal cot environment conditions are non-optimal for volatilization of antimony by S. brevicaulis, and that Sb2O3 in cot mattress PVC is not bioavailable. 6. Cot mattress isolates of S. brevicaulis also volatilized antimony (not encapsulated by PVC), whereas those of other filamentous fungi (Penicillium spp., Aspergillus niger, Aspergillus fumigatus, Alternaria sp.) and of bacteria (Bacillus spp.) did not. 7. The oxidation products of TMA may be the true determinants of toxicity for biogenic antimony gases produced in an aerobic environment.
Assuntos
Antimônio/metabolismo , Equipamentos para Lactente/microbiologia , Fungos Mitospóricos/metabolismo , Compostos Organometálicos , Morte Súbita do Lactente/etiologia , Antimônio/efeitos adversos , Antimônio/análise , Antimônio/química , Leitos , Biodegradação Ambiental , Meios de Cultura , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Recém-Nascido , Metilação , Cloreto de Polivinila/química , VolatilizaçãoRESUMO
1. Polyvinyl chloride (PVC) cot mattress covers from SIDS cases were investigated as potential sources of soluble (potentially ingestable) antimony in the cot environment. 2. Body fluids (urine, saliva) and proprietary domestic detergents/sterilizing fluids markedly enhanced leaching of antimony from PVC. Release of antimony was also enhanced at both low and high pH and by elevated temperature. The extent of antimony leaching did not correlate well with PVC content of this element. 3. These data do not support the assumption that postmortem analysis of antimony content proves exposure to gaseous antimony trihydride from mattress PVC. 4. Ingestion of antimony released from PVC could account for the high variability associated with reported detectable levels of antimony in liver from both SIDS and other infants. It could also explain suspected additional postnatal exposure to this element, which gives rise to elevated levels of Sb in the hair of some healthy infants.
Assuntos
Antimônio/análise , Equipamentos para Lactente , Cloreto de Polivinila/análise , Morte Súbita do Lactente/etiologia , Antimônio/efeitos adversos , Cabelo/química , Humanos , Concentração de Íons de Hidrogênio , Recém-Nascido , Fígado/química , Fatores de Risco , Saliva/químicaRESUMO
Recent reports of biovolatilisation of phosphorus and antimony by anaerobic bacteria and of leaching of phosphorus and antimony fire-retardant additives from PVC cot mattress covers, indicate that the polyurethane inner-foam of cot mattresses could be a site for generation of toxic gases of group 15 elements. A toxic gas hypothesis for sudden infant death syndrome (SIDS) involving polyurethane foam of cot mattresses was proposed and tested experimentally. Levels of antimony, phosphorus, arsenic and bismuth were determined at four sites for 44 SIDS and 50 control (no death) cot mattress foams. There was no evidence to suggest that the levels of these elements in cot mattress foam have a causal relation to SIDS. Leaching of antimony trioxide from PVC mattress covers could account for detectable levels of this element in 52% of the cot mattress samples analysed. Volatile forms of antimony, phosphorus, arsenic and bismuth was not detected in the headspace of mixed or monoseptic cultures of anaerobic bacteria containing polyurethane foam. Past microbial activity had given rise to involatile methylated species of antimony in some of the cot mattress foams tested (61%, n = 24). Abiotic oxidation of biogenic trimethylantimony together with physical adsorption of methylantimony forms to the polyurethane foam matrix could account for the apparent absence of "escaped" volatile antimony species in culture headspaces of incubation vial. There was no evidence to suggest that levels of trimethylantimony or total methylantimony forms in cot mattress foams have a causal relation to SIDS.
Assuntos
Bactérias Anaeróbias/metabolismo , Leitos/microbiologia , Gases/análise , Gases/toxicidade , Equipamentos para Lactente , Poliuretanos/química , Morte Súbita do Lactente/etiologia , Antimônio/análise , Arsênio/análise , Bismuto/análise , Cromatografia Gasosa-Espectrometria de Massas , Humanos , Lactente , Recém-Nascido , Compostos Organometálicos/análise , Fósforo/análise , Espectrofotometria Atômica , VolatilizaçãoRESUMO
UNLABELLED: Human exposure to cadmium (Cd) is associated with various diseases and high levels of Cd have been detected in Bangladeshi population warranting further research to identify the source of this exposure. In this study, Cd levels in 327 and 94 samples of Bangladeshi food and non-food samples, respectively, were determined using inductively coupled plasma mass spectrometry. This is the largest number of Bangladeshi food and nonfood samples investigated for their Cd content. High Cd levels were detected in leafy vegetables (mean 31 [SD 29]µg/kg). Of these vegetables, lal shak (Amaranthus tricolor) contained the highest Cd level (303 µg/kg [wet weight]; mean 100.5 [SD 95]µg/kg). Bangladeshi rice also showed significant concentration of Cd (mean 37.2 [SD 30]µg/kg). Of particular concern is the very high level of Cd detected in some puffed rice, which we attribute to the illegal practice of using urea for whitening the puffed rice. Tobacco leaves, which are commonly consumed during betel quid chewing by Bangladeshis, contain significant levels of Cd (mean 95 [SD 87]µg/kg). The total daily intake (TDI) of Cd from foods for Bangladeshis was estimated to be 34.55 µg/d. This is rather high when compared to the TDI of Cd for other populations. Our analysis reveals that this is mainly due to the very high intake of rice and vegetables, and lower consumption of animal products (which are low in Cd), by the Bangladeshis. We also determined the provisional maximum tolerable daily intake and target hazard quotients values for Cd. Clearly a more balanced diet is necessary to reduce the Cd intake in the Bangladeshi population, especially by reducing the very high intake of rice and certain leafy vegetables. Food manufacturing and agricultural practices needs to be altered to reduce the entry of Cd into the food chain. PRACTICAL APPLICATION: Exposure to high levels of Cd can be harmful to human health and this study provides a comprehensive analysis of Cd levels in a variety of food items from Bangladesh. The findings are of particular importance to consumers of Bangladeshi foods in both Bangladesh and in other countries. Data obtained will be valuable resources for food safety and regulatory bodies as our study suggests entry of Cd in foods through use of illegal chemicals in food manufacturing processes.
Assuntos
Cádmio/administração & dosagem , Cádmio/análise , Dieta/efeitos adversos , Contaminação de Alimentos , Adulto , Amaranthus/efeitos adversos , Amaranthus/química , Areca/efeitos adversos , Areca/química , Bangladesh , Intoxicação por Cádmio/prevenção & controle , Dieta/etnologia , Manipulação de Alimentos , Humanos , Dose Máxima Tolerável , Oryza/efeitos adversos , Oryza/química , Folhas de Planta/efeitos adversos , Folhas de Planta/química , Medição de Risco , Sementes/efeitos adversos , Sementes/química , Espectrofotometria Atômica , Nicotiana/efeitos adversos , Nicotiana/química , Tabaco sem Fumaça/efeitos adversos , Tabaco sem Fumaça/química , Reino Unido , Verduras/efeitos adversos , Verduras/químicaRESUMO
AIM: To investigate aerial release of bacteria from used cot mattresses and to assess factors that may influence this process. METHODS AND RESULTS: Movement on used mattresses, simulating that of an infant's head, significantly enhanced aerial release of naturally acquired bacteria from the polyurethane foams (total count data, P = 0.008; Staphylococcus aureus, P = 0.004) or from polyvinyl chloride covers (total count data, P = 0.001). Aerial release of naturally acquired bacteria from used cot mattresses showed high variability and was poorly correlated (R2 < or = 0.294) with bacterial cell density within the materials. In experiments involving inoculation of S. aureus and Escherichia coli onto the polyurethane of unused cot mattresses, aerial release of the species correlated well (R2 > or = 0.950) with inoculation density when simulated infant head movement was applied. Aerial release of these bacterial species from the material decreased with increase in width or aqueous content of the material, and was lower from polyurethane foam of a used cot mattress. CONCLUSIONS: Simulated infant movement and mattress related factors influence aerial release of bacteria from cot mattress materials. With simulated infant movement on cot mattress polyurethane foam, levels of airborne bacteria above the material are proportional to bacterial population levels inoculated onto the material. SIGNIFICANCE AND IMPACT OF THE STUDY: Cot mattresses harbouring relatively high levels of naturally acquired toxigenic bacteria, such as S. aureus, could pose a relatively high risk of infection to the infant's respiratory tract through increased aerial contamination. This has impact in the context of recent findings on cot mattress related risk factors for sudden infant death syndrome.
Assuntos
Poluição do Ar em Ambientes Fechados , Bactérias , Roupas de Cama, Mesa e Banho , Equipamentos para Lactente , Morte Súbita do Lactente/etiologia , Infecções Bacterianas/microbiologia , Infecções Bacterianas/transmissão , Contagem de Colônia Microbiana , Reservatórios de Doenças , Escherichia coli , Humanos , Lactente , Recém-Nascido , Movimento , Razão de Chances , Poliuretanos , Cloreto de Polivinila , Risco , Staphylococcus aureusRESUMO
AIMS: To compare growth and survival of selected bacteria implicated in sudden infant death syndrome (SIDS) on cot mattress polyurethane (PU) inner-foams and on different types of cot mattress cover materials. METHODS AND RESULTS: Escherichia coli, Staphylococcus aureus or Streptococcus pyogenes were inoculated onto swatches of new-unused cot mattress PU inner-foam and onto three types of cot mattress covers (polyvinyl chloride, cotton and polyester). The influence of inoculation cell density, relative humidity (RH) and temperature of incubation on survival was assessed by recovery of cells in 0.85% NaCl, with viable cell enumeration by plate counting on selective and differential media. Utilization of carbon and nitrogen sources within cot mattress PU was assessed by following growth on aqueous leachate from PU, and by colorimetric determination of aromatic amines. Good survival capability (>206 d) was shown by all three test species on PU inner-foam and on polyester mattress cover at high RH (75%), but only by Staph. aureus on PU at low RH (25%). Aqueous soluble material from PU foam supports bacterial growth; removal of aromatic amines from aqueous leachate from PU accompanies growth of Staph. aureus. CONCLUSIONS: Staphylococcus aureus has good survival capability on cot mattress PU foam, even at low RH. Soluble material within PU can serve as carbon and nitrogen sources for bacterial growth. SIGNIFICANCE AND IMPACT OF THE STUDY: Prolonged survival of Staph. aureus on PU at low RH could explain, in the context of the common bacterial toxins hypothesis, an increased risk of SIDS associated with used infant mattresses.
Assuntos
Roupas de Cama, Mesa e Banho/microbiologia , Viabilidade Microbiana , Morte Súbita do Lactente/etiologia , Aminoácidos Aromáticos/análise , Contagem de Colônia Microbiana , Colorimetria/métodos , Meios de Cultura , Escherichia coli/crescimento & desenvolvimento , Escherichia coli/isolamento & purificação , Humanos , Umidade , Lactente , Poliuretanos , Staphylococcus aureus/crescimento & desenvolvimento , Staphylococcus aureus/isolamento & purificação , Streptococcus pyogenes/crescimento & desenvolvimento , Streptococcus pyogenes/isolamento & purificação , TemperaturaRESUMO
Whole cells of Pseudomonas putida containing toluene dioxygenase were able to remove all detectable trichloroethylene (TCE) from assay mixtures. The capacity of cells to remove TCE was 77 microM/mg of protein with an initial rate of removal of 5.2 nmol/min/ng of protein. TCE oxidation resulted in a decrease in the growth rate of cultures and caused rapid cell death. Addition of dithiothreitol to assay mixtures increased the TCE removal capacity of cells by up to 67% but did not prevent TCE-mediated cell death. TCE induced toluene degradation by whole cells to a rate approximately 40% of that induced by toluene itself.
Assuntos
Oxigenases/metabolismo , Pseudomonas putida/enzimologia , Tricloroetileno/metabolismo , Biotransformação , Ditiotreitol , Oxirredução , Estresse Oxidativo , Pseudomonas putida/metabolismo , Tolueno/metabolismoRESUMO
Pseudomonas putida NCIMB 11767 oxidized phenol, monochlorophenols, several dichlorophenols and a range of alkylbenzenes (C1-C6) via an inducible toluene dioxygenase enzyme system. Biphenyl and naphthalene were also oxidized by this enzyme. Growth on toluene and phenol induced the meta-ring-fission enzyme, catechol 2,3-oxygenase, whereas growth on benzoate, which did not require expression of toluene dioxygenase, induced the ortho-ring-cleavage enzyme, catechol 1,2-oxygenase. Monochlorobenzoate isomers and 2,3,5-trichlorophenol were gratuitous inducers of toluene dioxygenase, whereas 3,4-dichlorophenol was a fortuitous oxidation substrate of the enzyme. The organism also grew on 2,4- and 2,5-dichloro isomers of both phenol and benzoate, on 2,3,4-trichlorophenol and on 1-phenylheptane. During growth on toluene in nitrogen-limited chemostat culture, expression of both toluene dioxygenase and catechol 2,3-oxygenase was positively correlated with increase in specific growth rate (0.11-0.74 h-1), whereas the biomass yield coefficient decreased. At optimal dilution rates, the predicted performance of a 1-m3 bioreactor supplied with 1 g nitrogen l-1 for removal of toluene was 57 g day-1 and for removal of trichloroethylene was 3.4 g day-1. The work highlights the oxidative versatility of this bacterium with respect to substituted hydrocarbons and shows how growth rate influences the production of competent cells for potential use as bioremediation catalysts.
Assuntos
Oxigenases/metabolismo , Pseudomonas putida/enzimologia , Biodegradação Ambiental , Poluentes Ambientais/metabolismo , Hidrocarbonetos/metabolismo , Pseudomonas putida/crescimento & desenvolvimento , Especificidade por Substrato , Tricloroetileno/metabolismoRESUMO
Toluene was oxidized by a mutant strain of Pseudomonas putida (strain NG1) to toluene Cis-Glycol (TCG). Product was accumulated in fed-batch cultures to concentrations (18-24 g/L) higher than hitherto achieved. In vitro activities of toluene dioxygenase from P. Putida NG1 were fivefold lower than that from the toluene-grown wild-type organism, whereas comparable activities of both catechol 2,3- and catechol 1,2-oxygenase were obtained; irreversible inhibition of toluene dioxygenase activity by TCG was shown in vitro. Ammonia deprivation during the production phase limited the growth of revertant organisms but had little effect on either the duration (25h) of the process or the final concentration of TCG achieved. The rate of glucose utilization decreased throughout the biotransformation and cell death accompanied the cessation of TCG accumulation in cultures. These changes were a consequence of TCG formation and a cooperative toxic effect was demonstrated for toluene and TCG. Adenylate energy charge values decreased from ca. 0.8 to 0.2 over the course of the biotransformation but were maintained above 0.5 in the absence of TCG. Similarly, cellular AMP levels increased dramatically during biotransformation, presumably as a consequence of RNA degradation, but were maintained at low levels in the absence of TCG. The results suggest that TCG is the mediate of a gradual deterioration in the state of the culture which leads to a loss of both in vivo and in vitro toluence dioxygenase activity and a marked decrease in culture viability.
RESUMO
Adaptation of methanol-grown Candida boidinii to ethanol utilization was accompanied by an increase in proteolytic activities, which behaved like known vacuolar enzymes. Degradation of alcohol oxidase protein was partially prevented by the serine proteinase inhibitor phenylmethanesulphonyl fluoride, but not by the carboxyl proteinase inhibitor pepstatin. Fractionation of cell-free extracts, by high-speed zonal centrifugation, of methanol-grown C. boidinii showed non-sedimentable and sedimentable proteolytic activities. Naturally occurring inhibitors of vacuolar proteinases were non-sedimentable. Fractionation of extracts prepared from methanol-grown cells which had been adapted to ethanol utilization for 5 h revealed significant changes in the sedimentability and distribution of proteolytic and acid phosphatase activities. These results suggest the possible involvement of a vacuolar process during alcohol oxidase degradation.