Oxygen Uptake Experiments Research Articles

Phylogenetic and functional analysis of cyanobacterial Cytochrome c6-like proteins.

All known photosynthetic cyanobacteria carry a cytochrome c 6 protein that acts transferring electrons from cytochrome b 6 f complex to photosystem I, in photosynthesis, or cytochrome c oxidase, in respiration. In most of the cyanobacteria, at least one homologue to cytochrome c 6 is found, the so-called cytochrome c 6B or cytochrome c 6C. However, the function of these cytochrome c 6-like proteins is still unknown. Recently, it has been proposed a common origin of these proteins as well as the reclassification of the cytochrome c 6C group as c 6B, renaming the new joint group as cytochrome c 6BC. Another homologue to cytochrome c 6 has not been classified yet, the formerly called cytochrome c 6-3, which is present in the heterocyst-forming filamentous cyanobacteria Nostoc sp. PCC 7119. In this work, we propose the inclusion of this group as an independent group in the genealogy of cytochrome c 6-like proteins with significant differences from cytochrome c 6 and cytochrome c 6BC, with the proposed name cytochrome c 6D. To support this proposal, new data about phylogeny, genome localisation and functional properties of cytochrome c 6-like proteins is provided. Also, we have analysed the interaction of cytochrome c 6-like proteins with cytochrome f by isothermal titration calorimetry and by molecular docking, concluding that c 6-like proteins could interact with cytochrome b 6 f complex in a similar fashion as cytochrome c 6. Finally, we have analysed the reactivity of cytochrome c 6-like proteins with membranes enriched in terminal oxidases of cyanobacteria by oxygen uptake experiments, concluding that cytochrome c 6D is able to react with the specific copper-oxidase of the heterocysts, the cytochrome c oxidase 2.

Oxidative Dissolution of Low-Grade Ni-Cu Ore and Impact on Flotation of Pentlandite

This paper investigated the effect of mineral surface oxidation on the floatability of Kevitsa low-grade Ni-Cu ore. Physicochemical measurements, ethylene diamine tetra acetic acid (EDTA) extraction, and oxygen uptake experiments were carried out with slurry and recycled process water samples obtained from the Kevitsa Cu-Ni sequential concentrator plant. The pH of recycled process water, copper flotation feed, and nickel flotation feed dropped by 0.7, 0.4, and 0.7 points, respectively, from May to July. The oxygen demand increased from recycled process water to the copper flotation feed, then dropped for the nickel flotation feed. The nickel flotation feed Redox potential (ORP) was lowest for July, while EDTA extractable metals increased from May to July. There was a 20% drop in nickel recoveries from May to July. Based on ORP measurements of the nickel flotation feed, good nickel flotation takes place in a moderately oxidizing (75–170 mV) and alkaline (9.2–9.7 pH) environment. Therefore, the ORP/pH of the nickel flotation feed is important to the nickel flotation. The results showed that at the Kevitsa plant, the grinding process is an electrochemically active environment, which, together with the incoming recycled process water quality, defines the degree of mineral surface oxidation for flotation. The increasing corrosiveness of the recycled process water increased mineral surface oxidation and depressed pentlandite flotation. Laboratory flotation experiments confirmed the observed poor plant flotation response when the corrosiveness of recycled process water increased. Total dissolved solids (TDS) was proven to be a reliable online parameter for the corrosiveness of the recycled process water and was inversely proportional to the pentlandite recovery. The findings of this study may help the plant develop ways to enable a timely response to changes in recycled process water quality to prevent harmful impacts on pentlandite flotation.

Synthesis and long‐term stability of acrylic acid and N, N‐methylene‐bis‐acrylamide radiation grafted polypropylene fibers

AbstractNew carboxyl ion‐exchange fibers prepared by acrylic acid (AA) and N, N‐methylene‐bis‐acrylamide (MBAA) monomer grafting to polypropylene (PP) fibers based on the method of γ‐rays pre‐irradiation in air followed by a Mohr's salt redox initiation of grafting reaction were studied. Grafting degree, homopolymer quantity, monomer conversion and grafting efficiency were determined. FTIR spectroscopy was used to establish the nature of the grafted fibers. The thermal stability of the materials was investigated based on DSC analysis in N2/O2 mixture. The long‐term stability during 6–8 years of storage was examined by the measurement of the tensile strength and the elongation at break of the fibers, and the chemical oxygen demand in water extracts as well as in the oxygen uptake experiments. The influence of some factors on the useful life of the grafted fibers was studied: a MBAA cross‐linking, exposure to day light, the Fe (III) residue presence, a grafting reaction temperature. It was found that small amounts of the adsorbed Fe (III) ions increase the deterioration rate of the grafted fibers, while small amounts of MBAA delay this negative process.

Diffusivity and Solubility of Oxygen in Solvents for Metal/Oxygen Batteries: A Combined Theoretical and Experimental Study

The solubility and the diffusion constant of oxygen in the electrolyte are crucial parameters in the kinetics of metal/oxygen batteries since they govern the discharge current that can be achieved. However, both parameters are often unknown or scatter widely in literature. Evaluating these parameters in typical solvents and electrolytes is therefore essential toward better quantitative understanding and improvement of metal/oxygen batteries. Using oxygen uptake experiments and molecular dynamics simulations, we compare theoretical and experimental approaches to determine oxygen diffusion coefficients in several molecular solvents that are commonly used. We report diffusion coefficients and Henry's law constants of oxygen for a series of glymes with different chain length and perfluorinated solvents, and discuss the benefit of using both experiment and simulation to determine these parameters. In all nine investigated solvents, the difference between simulated and measured diffusion coefficients is small in comparison to the magnitude of the coefficients. The methodology and the reported values for oxygen solubility and diffusivity form a solid starting point for further investigations into more complex electrolyte solutions. Ultimately, they might help to propel research on metal/oxygen batteries even further.

Interaction between Human Serum Albumin and antidiabetic compounds and its influence on the O2(1Δg)-mediated degradation of the protein

The complexity depicted by disease scenarios as diabetes mellitus, constitutes a very interesting field of study when drugs and biologically relevant components may be affected by such environments. In this report, the interaction between the protein Human Serum Albumin (HSA) and two antidiabetics (Andb), Gliclazide (Gli) and Glipizide (Glip) was studied through fluorescence and docking assays, in order to characterize these systems. On the basis that HSA and Andb can be exposed in vivo at high Reactive Oxygen Species (ROS) concentrations in diabetic patients, the degradative process of the protein free and bound to Andb, in presence of the species singlet molecular oxygen (O2(1Δg)), was evaluated. Fluorescence and docking assays indicated that Gli, as well as Glip bind to HSA on two sites, with binding constants values in the order of 104–105M−1. Likewise, docking assays revealed that the location of Gli or Glip on the protein may be the HSA binding sites II and III. Thermodynamic parameters showed that the interaction between HSA and Glip is a favored, enthalpically-controlled process. Oxygen uptake experiments indicated that Glip is less photooxidizable than Gli through a O2(1Δg)-mediated process. Besides, the protein–Andb binding produced a decrease in the overall rate constant for O2(1Δg) quenching as compared to the value for the free protein. This fact could be interpreted in terms of a reduction in the availability of Tyrosine residues in the bonded protein, with a concomitant decrease in the physical quenching deactivation of the oxidative species.

Insights into the Mechanism of Cumene Peroxidation Using Supported Gold and Silver Nanoparticles

Due to the considerable industrial implications, an in-depth study of cumene peroxidation using supported gold and silver nanoparticles was carried out to gain more insight into the mechanism of this reaction. Supported gold nanoparticles were found to efficiently catalyze the decomposition of cumene hydroperoxide with a selectivity of 25% at 80 °C when using gold supported on hydrotalcite (AuNP@HT), and 2-phenyl-2-propanol (i.e., cumyl alcohol) was the main product. Further, silver nanoparticles supported on hydrotalcite (AgNP@HT) converted cumene to cumene hydroperoxide at 80 °C with 80% selectivity. Both benchtop and oxygen-uptake experiments were used to probe the reaction mechanism and suggest that formation of a peroxyl radical-nanoparticle adduct is an important step in the peroxidation pathway and may be directly involved in the formation of the major cumyl alcohol product.

Photostability and photobactericidal properties of porphyrin-layered double hydroxide-polyurethane composite films.

The photostability and photobactericidal properties of PdTPPC (Pd(ii)-5,10,15,20-tetrakis(4-carboxyphenyl)porphyrin)-Zn2Al/PU (polyurethane) composite films have been studied in order to investigate their applicability as new photodynamic surfaces. These films comprise a PdTPPC porphyrin photosensitizer intercalated between the lamella of Zn2Al layered double hydroxide and dispersed (1 wt%) into a polyurethane matrix. The study of the photophysical behaviour shows that the Zn2Al LDH host enhances the chemical stability of the PdTPPC guest by minimizing photobleaching and quenching aggregation effects. The singlet oxygen production under irradiation of PdTPPC-Zn2Al/PU composite films is confirmed by the observation of an O2(1Δg) emission band centered at 1274 nm. Furthermore, the value of the rate constant kq for the PdTPPC phosphorescence quenching by oxygen kq = (8.2 ± 0.3) 10-2 s-1 Pa-1 indicates a slow diffusion of oxygen into and out of the PU polymer. In a second step, accelerating light ageing tests are conducted to determine the effect of singlet oxygen production on the chemical and mechanical stability of the PU matrix. Oxygen uptake experiments coupled with ATR-IR measurements indicate the probable formation of hydroxylated photoproducts but with no detrimental effects on the microstructure and the viscoelastic properties of the PU matrix as evidenced by dynamical mechanical analysis. Finally, in vitro preliminary antimicrobial tests show that PdTPPC-Zn2Al/PU composite films are able to inhibit S. aureus growth with no release of PdTPPC biocide from the PdTPPC-Zn2Al/PU composite film. We also observe a total inhibition of P. aeruginosa growth suggesting an efficacy against biofilm formation.

중합촉매 시스템이 폴리페닐렌에테르의 합성에 미치는 영향

Poly(2,6-dimethyl-1,4-phenylene ether) (PPE) was synthesized using or CuCl catalyst with various amounts of ligand and base in several different solvent systems. CuCl/1-methylimidazole/ammonium hydroxide was found to be an effective catalyst system which showed the highest polymer yield and molecular weight. The effects of catalyst/monomer ratio, different amine ligands, and the content of mono-functional reagent 2,4,6-trimethylphenol (TMP) additive on the polymer yield and molecular weight were investigated. Among the co-solvent systems used in this polymerization, chloroform/methanol 9/1(v/v) gave the highest polymer yield and molecular weight ( 55 K, 92 K, PDI 1.7). The catalytic activity between CuCl and CuI was compared by oxygen-uptake experiments and the formation of sideproduct, 5,5'-tetramethyl-4,4'-diphenoquinone (DPQ), was analyzed by ultraviolet spectroscopy.

Potassium channel‐oxidative phosphorylation relationship in durum wheat mitochondria from control and hyperosmotic‐stressed seedlings

Durum wheat mitochondria (DWM) possess an ATP-inhibited K(+) channel, the plant mitoK(ATP) (PmitoK(ATP) ), which is activated under environmental stress to control mitochondrial ROS production. To do this, PmitoK(ATP) collapses membrane potential (ΔΨ), thus suggesting mitochondrial uncoupling. We tested this point by studying oxidative phosphorylation (OXPHOS) in DWM purified from control seedlings and from seedlings subjected both to severe mannitol and NaCl stress. In severely-stressed DWM, the ATP synthesis via OXPHOS, continuously monitored by a spectrophotometric assay, was about 90% inhibited when the PmitoK(ATP) was activated by KCl. Contrarily, in control DWM, although PmitoK(ATP) collapsed ΔΨ, ATP synthesis, as well as coupling [respiratory control (RC) ratio and ratio between phosphorylated ADP and reduced oxygen (ADP/O)] checked by oxygen uptake experiments, were unaffected. We suggest that PmitoK(ATP) may play an important defensive role at the onset of the environmental/oxidative stress by preserving energy in a crucial moment for cell and mitochondrial bioenergetics. Consistently, under moderate mannitol stress, miming an early stress condition, the channel may efficiently control reactive oxygen species (ROS) generation (about 35-fold from fully open to closed state) without impairing ATP synthesis. Anyway, if the stress significantly proceeds, the PmitoK(ATP) becomes fully activated by decrease of ATP concentration (25-40%) and increase of activators [free fatty acids (FFAs) and superoxide anion], thus impairing ATP synthesis.

Two-Phase Partitioning Bioreactors Operating with Polymers Applied to the Removal of Substituted Phenols

Significant improvement in biodegradation performance has been demonstrated arising from the reduction of cytotoxicity provided by the sequestering of 4-nitrophenol (4NP) within Hytrel polymer beads added to a two-phase partitioning bioreactor (TPPB) operating in sequencing batch reactor (SBR) mode. This reduced toxicity is particularly apparent as the feed substrate concentration is increased; in fact it was shown that at a feed of 1000 mg/L 4NP, the inhibitory effect of the substrate completely prevents degradation from occurring in a single-phase system, whereas at only a 5% polymer loading, rapid and compete biodegradation is achieved. Different polymer/aqueous phase ratios were used to detoxify varying feed concentrations, and degradation rates were enhanced through the use of increased polymer loadings. As demonstrated in oxygen uptake experiments, the addition of polymers also reduces the maximum demand for oxygen, relative to single-phase operation, and smoothes the demand for oxygen throughout the degradation process. Polymer regeneration has also been further characterized by quantifying the number of methanol washes required to achieve satisfactory 4NP residuals, and the addition of a small amount of cosolvent has been shown to dramatically increase the rate of bioregeneration to produce beads ready for reuse.

A Comparative Kinetic and Mechanistic Study Between Tetrahydrozoline and Naphazoline Toward Photogenerated Reactive Oxygen Species

Kinetic and mechanistic aspects of the vitamin B2 (riboflavin [Rf])-sensitized photo-oxidation of the imidazoline derivates (IDs) naphazoline (NPZ) and tetrahydrozoline (THZ) were investigated in aqueous solution. The process appears as important on biomedical grounds, considering that the vitamin is endogenously present in humans, and IDs are active components of ocular medicaments of topical application. Under aerobic visible light irradiation, a complex picture of competitive interactions between sensitizer, substrates and dissolved oxygen takes place: the singlet and triplet ((3)Rf*) excited states of Rf are quenched by the IDs: with IDs concentrations ca. 5.0 mM and 0.02 mM Rf, (3)Rf* is quenched by IDs, in a competitive fashion with dissolved ground state oxygen. Additionally, the reactive oxygen species: O(2)((1)Delta(g)), O(2)(*-), HO(*) and H(2)O(2), generated from (3)Rf* and Rf(*-), were detected with the employment of time-resolved methods or specific scavengers. Oxygen uptake experiments indicate that, for NPZ, only H(2)O(2) was involved in the photo-oxidation. In the case of THZ, O(2)(*-), HO(*) and H(2)O(2) were detected, whereas only HO(*) was unambiguously identified as THZ oxidative agents. Upon direct UV light irradiation NPZ and THZ generate O(2)((1)Delta(g)), with quantum yields of 0.2 (literature value, employed as a reference) and 0.08, respectively, in acetonitrile.

Oxidative dissolution of bornite by Acidithiobacillus ferrooxidans

The oxidation of finely ground (−200 μm) bornite (Cu 5FeS 4) by Acidithiobacillus ferrooxidans was evaluated in oxygen uptake and shake flasks experiments. The oxidation was a net acid-consuming reaction. Residual bornite was not detected by X-fray diffraction in solids after 2 days of contact in acid leach solution, indicating that the chemical and biological oxidation of bornite was relatively fast. Virtually 100% of copper solubilization was achieved in A. ferrooxidans cultures with or without ferrous iron, while in abiotic controls the copper extraction was around 30%. Bornite was not oxidized by Acidithiobacillus thiooxidans in respirometric or shake flasks experiments. Covellite (CuS) was detected as a secondary phase under all experimental conditions. Sulfur and jarosite were formed only in the presence of A. ferrooxidans.

Elucidation of the ipso-substitution mechanism for side-chain cleavage of alpha-quaternary 4-nonylphenols and 4-t-butoxyphenol in Sphingobium xenophagum Bayram.

Recently we showed that degradation of several nonylphenol isomers with alpha-quaternary carbon atoms is initiated by ipso-hydroxylation in Sphingobium xenophagum Bayram (F. L. P. Gabriel, A. Heidlberger, D. Rentsch, W. Giger, K. Guenther, and H.-P. E. Kohler, J. Biol. Chem. 280:15526-15533, 2005). Here, we demonstrate with 18O-labeling experiments that the ipso-hydroxy group was derived from molecular oxygen and that, in the major pathway for cleavage of the alkyl moiety, the resulting nonanol metabolite contained an oxygen atom originating from water and not from the ipso-hydroxy group, as was previously assumed. Our results clearly show that the alkyl cation derived from the alpha-quaternary nonylphenol 4-(1-ethyl-1,4-dimethyl-pentyl)-phenol through ipso-hydroxylation and subsequent dissociation of the 4-alkyl-4-hydroxy-cyclohexadienone intermediate preferentially combines with a molecule of water to yield the corresponding alcohol and hydroquinone. However, the metabolism of certain alpha,alpha-dimethyl-substituted nonylphenols appears to also involve a reaction of the cation with the ipso-hydroxy group to form the corresponding 4-alkoxyphenols. Growth, oxygen uptake, and 18O-labeling experiments clearly indicate that strain Bayram metabolized 4-t-butoxyphenol by ipso-hydroxylation to a hemiketal followed by spontaneous dissociation to the corresponding alcohol and p-quinone. Hydroquinone effected high oxygen uptake in assays with induced resting cells as well as in assays with cell extracts. This further corroborates the role of hydroquinone as the ring cleavage intermediate during degradation of 4-nonylphenols and 4-alkoxyphenols.

Aerobic conversion of dimethyl sulfide and hydrogen sulfide by Methylophaga sulfidovorans: implications for modeling DMS conversion in a microbial mat

Methylophaga sulfidovorans is an obligately methylotrophic, DMS-oxidizing organism, isolated from microbial mat sediment. DMS and H2S, both present in marine microbial mats, can be used as energy sources by this organism. In batch cultures of M. sulfidovorans, sequential H2S and DMS utilization occurred. In energy-limited continuous cultures, with DMS, methanol and H2S as substrates, mixotrophic growth of M. sulfidovorans was observed, showing that at low concentrations these substrates can be used simultaneously. Oxygen and H2S uptake experiments showed that the critical concentration at which sulfide inhibition of DMS oxidation occurred was between 15 and 40 μmol l−1. Also in crude enrichments of DMS oxidizers a decrease of 50% in DMS-oxidizing capacity for about 200 μmol l−1 H2S was observed. The new physiological data obtained with the pure cultures of M. sulfidovorans were incorporated in a compartment model of a microbial mat and gave improved predictions of DMS profiles and DMS emissions from the mat, both when phototrophic activity is present (day) and when it is absent (night).

Simultaneous oxygen uptake and imaging chemiluminescence measurements

There are several studies that make a comparison between chemiluminescence and oxygen uptake. Although, there are several reasons to perform this comparison on the same sample at the same conditions, these studies were done by comparing results from separate oxygen uptake and chemiluminescence experiments. It is also postulated that the shape of the chemiluminescence and oxygen uptake curves is not related to the kinetics of the degradation chemistry but to the oxidizing area. To be able to measure the luminescence and the oxygen uptake at the same sample under exactly the same conditions an apparatus is developed in which both measurements can be done simultaneously. To be able to get information on the oxidizing area too, this apparatus is based on imaging chemiluminescence. The induction times determined with chemiluminescence and oxygen uptake are about comparable, although at lower temperatures the induction time according to oxygen uptake is shorter, which might be due to a decrease of the sensitivity of chemiluminescence with temperature. For stabilized and unstabilized polypropylene it was shown that the luminescence curve is related to the amount of material degrading and that the shape of the oxygen uptake and luminescence curves is not related to the kinetics of the degradation chemistry.

Improved method for isolation of coupled mitochondria of Araucaria angustifolia (Bert.) O. Kuntze

A method for the isolation of coupled mitochondria from the callus of Araucaria angustifolia is described for the first time. Mitochondria were isolated from embryogenic callus of A. angustifolia. They were metabolically active, able to sustain oxidative phosphorylation as shown by respiratory control ratio values, which were about 2.4 when respiring on succinate as substrate. Oxygen uptake experiments, using freeze-thawed disrupted mitochondria, showed the presence of alternative rotenone-insensitive NAD(P)H dehydrogenases, which were stimulated by Ca2+. The procedure now described for the isolation of A. angustifolia mitochondria is an important new tool, allowing the investigation of mitochondrial bioenergetics and metabolism and physiology of plants.

Efeito da concentração inicial de oxigênio no coeficiente de aeração para experimentos de DBO de longo prazo

Long-term incubations of biochemical oxygen demand (BOD) were performed to determine the aeration coefficient derived from oxygen uptake experiments. The BOD bottles were prepared with distilled water and the incubation occurred in the dark, under controlled temperature (19.4ºC). The bottles were submitted to two different treatments based on the initial dissolved oxygen (DO) concentrations (Treatment 1: 1.43mg L-1 and Treatment 2: 7.67mg L-1). The DO concentrations were measured with an OD meter during 52 days. The results were fitted to a first-order kinetics model. The BOD bottles showed an increase in DO concentration for both treatments and the mean aeration coefficient (ka) was 0.065 a day-1. The fittings pointed to some dispersion in the ka values, and the initial concentration of DO in the BOD bottles does not interfere in the aeration process, being a random process

Two mechanisms for oxidation of cytosolic NADPH by Kluyveromyces lactis mitochondria.

Null mutations in the structural gene encoding phosphoglucose isomerase completely abolish activity of this glycolytic enzyme in Kluyveromyces lactis and Saccharomyces cerevisiae. In S. cerevisiae, the pgi1 null mutation abolishes growth on glucose, whereas K.lactis rag2 null mutants still grow on glucose. It has been proposed that, in the latter case, growth on glucose is made possible by an ability of K. lactis mitochondria to oxidize cytosolic NADPH. This would allow for a re-routing of glucose dissimilation via the pentose-phosphate pathway. Consistent with this hypothesis, mitochondria of S. cerevisiae cannot oxidize NADPH. In the present study, the ability of K. lactis mitochondria to oxidize cytosolic NADPH was experimentally investigated. Respiration-competent mitochondria were isolated from aerobic, glucose-limited chemostat cultures of the wild-type K. lactis strain CBS 2359 and from an isogenic rag2Delta strain. Oxygen-uptake experiments confirmed the presence of a mitochondrial NADPH dehydrogenase in K.lactis. This activity was ca. 2.5-fold higher in the rag2Delta mutant than in the wild-type strain. In contrast to mitochondria from wild-type K. lactis, mitochondria from the rag2Delta mutant exhibited high rates of ethanol-dependent oxygen uptake. Subcellular fractionation studies demonstrated that, in the rag2Delta mutant, a mitochondrial alcohol dehydrogenase was present and that activity of a cytosolic NADPH-dependent 'acetaldehyde reductase' was also increased. These observations indicate that two mechanisms may participate in mitochondrial oxidation of cytosolic NADPH by K. lactis mitochondria: (a) direct oxidation of cytosolic NADPH by a mitochondrial NADPH dehydrogenase; and (b) a two-compartment transhydrogenase cycle involving NADP(+)- and NAD(+)-dependent alcohol dehydrogenases.

Importance of weathering factors other than UV radiation and temperature in outdoor exposure

Results of outdoor exposure experiments on HALS-stabilized low-density polyethylene blown films at well-known weathering sites in southern France and Miami, Florida could not be explained on the basis of meteorological data on factors generally held responsible for polymer degradation in outdoor applications. Unstabilized and non-HALS-stabilized LDPE films on the contrary showed the behaviour expected under the experimental conditions. Oxygen uptake experiments with concomitant analysis of degradation products and monitoring of mechanical behaviour during natural ageing in closed borosilicate glass vessels did not reveal significantly different degradation chemistries in the two climates. Comparison of weathering behind quartz glass and direct weathering shed light on the probable reason for the unexpected lifetime ranking of HALS-stabilized LDPE films in both exposure regions. Additive formulations show specific responses to different climates. This has implications for durability assessment and stabilization strategies.

Bioconversion of Terephthalate by Resting Cells of Alkaliphilic Dietzia sp. Strain GS-1.

Disodium terephthalate (DT) was degraded by the resting cells of alkaliphilic Dietzia sp. strain GS-1. The degradation occurred at pH 7.0-10.0. The optimal pH for the degradation was around 8.0. Oxygen uptake experiments with the resting cells showed that oxygen was utilized for degrading DT by strain GS-1. Thin layer chromatography (TLC) analysis revealed three degradation products in the resting cells reaction at pH 10. The products absorbed light at around 265 nm and were likely aromatic compound having hydroxyl and/or carboxyl group. The resting cells reaction was strongly inhibited by a,a'-dipyridyl, suggesting DT-degrading enzyme is likely metalloenzyme. The cell-free extracts degraded DT at the velocity of 0.885 μM min - 1 mg-protein - 1 .