Cloning of DNA from a Rhodococcus strain conferring the ability to decolorize sulfonated azo dyes

Azo dyes are recalcitrant pollutants. Two sulfonated azo dyes, Orange II and Amido black, are effectively decolorized by certain nocardioform strains of the genus Rhodococcus. A mutant of one of these strains was isolated which had lost azo-dye decolorizing ability and the strain was used to clone DNA conferring this ability, by screening a BclI library constructed from DNA of a decolorizing strain. The relevant genetic information was located on a 6.3-kb fragment of DNA.     

Kinetic and thermodynamic characterization of the functional properties of a hybrid versatile peroxidase using isothermal titration calorimetry: Insight into manganese peroxidase activation and lignin peroxidase inhibition

Isothermal titration calorimetry (ITC) was developed for measuring lignin peroxidase (LiP) and manganese peroxidase (MnP) activities of versatile peroxidase (VP) from Bjerkandera adusta. Developing an ITC approach provided an alternative to colorimetric methods that enabled reaction kinetics to be accurately determined. Although VP from Bjerkandera adjusta is a hybrid enzyme, specific conditions of [Mn+2] and pH were defined that limited activity to either LiP or MnP activities, or enabled both to be active simultaneously. MnP activity was found to be more efficient than LiP activity, with activity increasing with increasing concentrations of Mn+2. These properties of MnP were explained by a second metal binding site involved in homotropic substrate (Mn+2) activation. The activation of MnP was also accompanied by a decrease in both activation energy and substrate (Mn) affinity, reflecting a flexible enzyme structure. In contrast to MnP activity, LiP activity was inhibited by high dye (substrate) concentrations arising from uncompetitive substrate inhibition caused by substrate binding to a site distinct from the catalytic site. Our study provides a new level of understanding about the mechanism of substrate regulation of catalysis in VP from B. adjusta, providing insight into a class of enzyme, hybrid class II peroxidases, for which little experimental data is available.     

藻对偶氮染料降解作用的研究

本实验系统地研究了藻对偶氮染料的降解作用。实验结果表明,藻对多种偶氮染料具有一定的降解作用,其降解程度与染料的结构和藻的种类等因素有关。通过降解机理的研究表明,,藻能够利用偶氮还原酶,作用于偶氮染料的偶氮双键,使其断裂,形成芳香胺类中间产物。藻还可以进一步利用芳香胺类物质。本研究结果启示,应重新评价藻在氧化塘降解有机物质过程中所起的作用,即在氧化塘中,藻不仅起供氧作用,还能够直接参与有机物质的降解。     

The effect of Azo-dyes on glioblastoma cells in vitro

Despite the multidisciplinary standard treatment of glioblastoma (GB) consisting of maximal surgical resection, followed by radiotherapy (RT) plus concomitant chemotherapy with temozolomide (TMZ), the majority of patients experience tumor progression and almost universal mortality. In recent years, efforts have been made to create new agents for GB treatment, of which azo-dyes proved to be potential candidates, showing antiproliferative effects by inducing apoptosis and by inhibiting different signaling pathways. In this study we evaluated the antiproliferative the effect of six azo-dyes and TMZ on a low passage human GB cell line using MTT assay. We found that all compounds proved antiproliferative properties on GB cells. At equimolar concentrations azo-dyes induced more cytotoxic effect than TMZ. We found that Methyl Orange required the lowest IC₅₀ for 3 days of treatment (26.4684 μM), whilst for 7 days of treatment, two azo dyes proved to have the highest potency: Methyl Orange IC₅₀ = 13.8808 μM and Sudan I IC₅₀ = 12.4829 μM. The highest IC₅₀ was determined for TMZ under both experimental situations. Conclusions: Our research represents a novelty, by offering unique valuable data regarding the azo-dye cyototoxic effects in high grade brain tumors. This study may focus the attention on azo-dye agents that may represent an insufficient exploited source of agents for cancer treatment.