Strategies for the design and operation of enzymatic reactors for the degradation of highly and poorly soluble recalcitrant compounds

The presence of recalcitrant compounds in both wastewaters and soils is an important environmental problem. Oxidative enzymes from white-rot fungi have been successfully utilised for the in vitro degradation of xenobiotics, such as the azo dye Orange II and the polycyclic aromatic hydrocarbon anthracene (compounds with high and low solubilities, respectively). Two different reactor configurations are proposed: (i) an enzymatic membrane reactor for the treatment of soluble compounds, consisting of a continuous stirred tank reactor coupled to an ultrafiltration membrane to facilitate the retention and recycling of enzyme; and (ii) a two-phase enzymatic reactor for the degradation of poorly soluble compounds, consisting of an immiscible solvent, which contains the contaminant at high concentrations, and the aqueous phase containing the enzyme and cofactors involved in the catalytic cycle. In this paper, factors affecting the design and operation of both systems are discussed, and experimental results concerning the efficiency and stability of the processes are presented.     

fliY基因失活减弱沙福芽孢杆菌ST7菌株的锰氧化能力

【背景】沙福芽孢杆菌ST7菌株具有较强的锰氧化能力,但其分子机制不清楚。【目的】着重研究鞭毛马达开关蛋白基因(fliY)对沙福芽孢杆菌锰氧化能力的影响。【方法】根据同源重组原理,以沙福芽孢杆菌ST7菌株为起始菌株,构建fliY基因敲除的突变株ΔfliY,测定菌落迁徙、细菌生物膜和锰氧化率等,研究fliY基因突变后菌株的运动能力、生物膜生成和锰氧化能力是否发生变化。【结果】经克隆测序,证实突变株ΔfliY中fliY基因的后半段被卡纳霉素抗性基因取代,fliY基因失活;与野生型菌株ST7相比,突变株ΔfliY在全营养的LB培养基中生长变化不大,但在含锰的PYCM培养基中,突变株的生长速度减慢、菌落较小、生物膜生成量显著下降,运动性和锰氧化能力分别下降65%和20%。【结论】fliY基因不仅影响菌株的生长和运动,而且参与细菌的趋化和锰氧化等生物学过程。     

Effects of Zn2+ and Cu2+ on growth,lignin degradation and ligninolytic enzymes in Phanerochaete chrysosporium

The influence of Zn²⁺ (6.0 × 10^–3 –18.0 × 10^–3 M) and Cu²⁺ (4 × 10^–4 –1.2 × 10^–4 M) in the basal medium on mycelial growth (dry weight), activities of lignin peroxidase (Lip), manganese peroxidase (Mnp), solubilization, and mineralization (¹⁴CO₂ evolution) of lignin during a period of 3 weeks was studied in Phanerochaete chrysosporium strain MTCC-787. Highest mycelial growth was obtained at 0.6 M Zn²⁺ and 0.4 M Cu²⁺ levels. Enzyme activities were found to increase up to the highest levels of both the trace elements. However, Zn²⁺ had a relatively more stimulatory effect on Lip production and the reverse was true in case of Cu²⁺. [¹⁴C]Lignin solubilization was also promoted by higher levels of both trace elements. Mineralization of [¹⁴C]lignin was optimal at 6.0 M Zn²⁺ and 1.2 M Cu²⁺. The stimulatory effect of Zn²⁺ on Lip production was correlated with higher rates of [¹⁴C]lignin mineralization.     

Effect of Chronic Manganese Treatment on Adenosine Tissue Levels and Adenosine A2a Receptor Binding in Diverse Regions of Mouse Brain

In the present study the effects of chronic manganese (Mn) treatment on adenosine A_2a receptor binding in mouse brain have been assessed. Male albino mice were divided in two groups: In the Mn-treated group, the animals were injected intraperitoneally (i.p.) with MnCl₂ (5 mg/kg/day) five days per week during 9 weeks; in the control group, they were injected likewise with a saline solution. A significant decrease of the Kd without alteration of Bmax in the cerebellum and, an increase of the Kd and Bmax in hippocampus of mice treated with Mn were found. Also, an increase of Kd in frontal cortex was observed. The binding parameters in caudate nucleus, olfactory bulb and hypothalamus were not altered by Mn. A significant decrease in the adenosine concentration in caudate nucleus, olfactory bulb and hypothalamus, without significant changes in hippocampus, frontal cortex and cerebellum was also detected. These findings suggest that chronic administration of Mn could affect adenosine receptor function and turnover, depending on the brain region analyzed.     

Alterations in plasma essential trace elements selenium, manganese, zinc, copper, and iron concentrations and the possible role of these elements on oxidative status in patients with childhood asthma

The aim of the present study is to evaluate the status of plasma essential trace element selenium (Se), manganese (Mn), copper (Cu), zinc (Zn), and iron (Fe) concentrations and the effect of these elements on oxidative status in patients with childhood asthma. Plasma Se, Mn, Cu, and Zn concentrations were determined by atomic absorption spectrophotometry (AAS) and Fe concentrations, malondialdehyde (MDA), and total antioxidant capacity (TAC) were determined by the colorimetric method. The plasma MDA/TAC ratio was calculated as an index of oxidative status. Plasma albumin levels were measured to determine nutritional status. Plasma Fe concentrations, MDA levels and the MDA/TAC ratio were significantly higher (p<0.001, p<0.001, and p<0.01, respectively) and Se and Mn concentrations and TAC were lower (p<0.01, p<0.05, and p<0.01, respectively) in patients when compared to the healthy subjects. Plasma Zn, Cu, and albumin levels were not found to be significantly different in patients and controls (p>0.05). There were positive relationships between plasma MDA and Fe (r=0.545, p<0.001) and TAC and Se (r=0.485, p<0.021), and a negative correlation between TAC and MDA values (r= −0.337, p<0.031) in patients with childhood asthma. However, there was no correlation between these trace elements and albumin content in patient groups. These observations suggest that increased Fe and decreased Se concentrations in patients with childhood asthma may be responsible for the oxidant/antioxidant imbalance.     

Novel Rh-Mn mixed-metal carbonyl cluster containing μ3 and μ-bridging carbyne ligands, [(μ3-CC6H5)(μ-CC6H5)Rh2Mn2Cp2(μ-CO)3(CO)3]: structure, electrochemistry and catalytic properties

The cationic carbyne complex [Cp(CO)₂MnCC₆H₅]BBr₄ (1) reacts with PPN[Rh(CO)₄] (2) to give the title cluster [(μ₃-CC₆H₅)(μ-CC₆H₅) Rh₂Mn₂Cp₂(μ-CO)₃(CO)₃] (3) whose structure has been determined by X-ray diffraction. The electrochemical properties of 3 have been investigated using cyclic voltammetric method. At 60 °C and 2.0 MPa of initial total CO/H₂ (1:1) pressure, the catalytic activity of 3 towards hydroformylation of styrene has also been checked.     

Inorganic salts influence IAA ionization and adventitious rhizogenesis in Pongamia pinnata

The basal cut end of coppice shoot cuttings of Pongamia pinnata was treated for 24 h with 0 (water treated control) or 5.0 mmol/L of KMnO4, KCI, and KH2PO4 or 2.5 mmol/L of K2HPO4 and K2SO4. Inorganic salts of P, S, Cl and Mn significantly influenced IAA ionization and adventitious rhizogenesis. P and S salts had lower IAA ionization potential, but more pronounced effect on adventitious rhizogenesis than Cl and Mn salts. The linear regression analysis also established negative correlations between salt induced IAA ionization with various characteristics of adventitious rhizogenesis such as sprouting (r = -0.83, p < 0.05), rooting (r = -0.82, p < 0.05), root number (r = -0.95, p < 0.01), and root length (r = -0.80, p < 0.1). The implication of IAA ionization in adventitious rhizogenesis has been discussed and the possible role of inorganic salts therein suggested.     

Decolorization of azo dyes using Basidiomycete strain PV 002

Summary Basidiomycete PV 002, a recently isolated white-rot strain from decomposed neem waste displayed high extracellular peroxidase and rapidly decolorized azo dyes. In this study, the optimal culture conditions for efficient production of ligninolytic enzymes were determined with respect to carbon and nitrogen. An additional objective was to determine the efficiency of PV 002 to degrade the azo dyes. White-rot strain PV 002 efficiently decolorized Ranocid Fast Blue (96%) and Acid Black 210 (70%) on day 5 and 9 respectively under static conditions. The degradation of azo dyes under different conditions was strongly correlated with the ligninolytic activity. The optimum growth temperature of strain PV 002 was 26 °C and pH 7.0.     

Manganese Oxidation by Bacterial Isolates from the Indian Ridge System

The abundance and activity of culturable manganese-oxidizing bacteria were assessed from near-bottom water samples of the tectonically active Carlsberg Ridge. Retrievable counts as colony forming units (CFU) on dilute nutrient agar medium (dilNA = 2 gm l⁻¹ nutrient broth+2% agar) and on dilNA supplemented with 1, 2 and 3 mM MnCl₂·4H₂O were in the order of 10⁶ CFU l⁻¹. Retrievability of heterotrophs ranged from non-detectable levels (ND) to 2.82 × 10⁶ CFU l⁻¹. The retrievable counts on Mn amended dilNA ranged from ND to 3.21× 10⁶, 1.47 × 10⁶ and 1.45 × 10⁶ CFU l⁻¹ on 1, 2 and 3 mM, respectively. About 87% of the Mn tolerant isolates (n = 39) showed taxonomic affinities to Pseudomonas I and II sp. Two representative strains CR35 and CR48 (CR–Carlsberg Ridge) isolated on manganese-supplemented media were tested for their ability to tolerate a range of Mn amendments from 1 nM to 100 mM in terms of growth and respiration. CR35 represents 66% of the total CFU (3.04 × 10⁶ CFU l⁻¹), while CR48 represented only 6% of the total CFU (1.05 × 10⁶ CFU l⁻¹). The colonies of these two isolates were dark brown in color suggesting precipitation of Mn as oxide. Tests for the effect on growth and respiration were conducted in media simulating heterotrophic (amended with 0.01% glucose) and lithotrophic (unamended) conditions. Maximum stimulation in growth and respiration of CR35 occurred at 100 μM Mn both in unamended and amended media. At levels of Mn greater than 100 μM the counts decreased steadily. Total respiring cells of CR48 were stimulated to a maximum at 1 μM Mn in unamended medium and 1 nM in amended medium. Total cells counts for the same decreased beyond 100 μM Mn in unamended and 1 nM in amended medium. The isolates were tested for their ability to oxidize Mn ammendments from 1 μM to 10 mM Mn. At the end of a 76-day incubation period, there was evidence of manganese oxide precipitation at high Mn concentrations (≥1 mM) as a dark brown coloration on the sides of culture tubes. Highest Mn oxidation rates were observed at 10 mM Mn(II) concentration with CR35 oxidizing 27 and 25 μM Mn day⁻¹ in unamended and amended condition, respectively. CR48 oxidized Mn at the rate of 26 μM Mn day⁻¹ in unamended medium and 35 μM Mn day⁻¹ in amended medium. Scanning electron microscope (SEM) observations of both isolates revealed free-living cells in clustered matrices ≈2 μm diameter. Energy dispersive spectrum of the cell matrix of CR35 cultured in 1 mM Mn detected 30% Mn, while the cell aggregates of CR48 harbored 7–10% Mn. The relatively high specific activity of these mixotrophic bacteria under relatively oligotrophic conditions suggests that they may be responsible for scavenging dissolved Mn from the Carlsberg Ridge waters and could potentially participate in oxidation.