Dye decolorisation by laccase entrapped in copper alginate

A novel immobilisation system was developed for dye decolorisation using laccase produced by Ganoderma sp. KU-Alk4. The enzyme showed high efficiency in dye decolorisation when entrapped in Cu–Al and Cu-alginate beads. The former gave the highest activity but the enzyme activity survived longer in the latter. An experimental design of two 3 × 3 Latin Square experiments was applied to evaluate the effects of three different alginate compositions (low, intermediate and high mannuronate), concentration of alginate, (1.5, 3.0 and 4.5% w/v) and concentration of cross-linking agent, CuSO₄ (0.075, 0.15 and 0.225 M) on the decolorisation of indigo carmine dye and residual laccase activity in beads. The most significant factor for residual activity was the concentration of the cross-linking agent (P < 0.05) followed by alginate composition (P < 0.1). Increasing the alginate concentration resulted in only small increase in the dye decolorisation. However, higher laccase activity remained in 3.0% w/v alginate beads. Maximal dye decolorisation was achieved when 3.6% w/v low mannuronate alginate and 0.15 M CuSO₄ was used. Optimal conditions were confirmed in an extended experimental run. Results are presented from 9 successive batch runs over 12 days, reaching 96% removal of the dye (216 mg/l).     

Flexible polymer TiO2 modified film photocatalysts active in the photodegradation of azo-dyes in solution

This review summarizes the recent developments of thin film polymer coated photocatalysis with TiO₂ mediating the discoloration/degradation of the azo-dye Orange II under light irradiation. The stable anchoring of TiO₂ on non-heat resistant but chemically inert flexible polymer films is described. The nature of the polymer films used, the pretreatment of the film for the TiO₂ loading and the testing of the photocatalytic activity are addressed for different inert polymer films not having the conventional functional surface groups to bind TiO₂. The discoloration of Orange II in the presence of LDPE/TiO₂ is completed in about 10 h. This is a significantly longer times than the one observed for the same process when Tedlar/TiO₂ and Parylene/TiO₂ were used in the dye discoloration process. This points out to specific effects particular to each the polymer support used to graft the photoactive TiO₂ particles.     

Benzoxazinone — kanamycin derivative: a new fluorescent probe for flow cytometry analysis of bacteria (Agrobacterium tumefaciens)

Abstract A new polycation fluorescent dye (BVC-kinamycin-conjugate) has been synthesized and used to detect alive bacteria by flow cytometry. This fluorescent chromophore has the noteworthy property of being excited at the same wavelength as fluoresceinylated conjugates (488 nm) and to show a much longer emission wavelength (616 nm) than fluoresceinylated derivates (520 nm); furthermore its fluorescence intensity is not quenched at low pH in contrast with fluorescein. In such conditions, bacteria can easily be detected with cytofluorimeter equipped with a single excitation wavelength beam. The binding of fluoresceinylated lectins and antibodies onto a strain of Agrobacterium tumefaciens has been studied by this method.     

Biochemical and molecular characterization of a novel laccase from selective lignin-degrading white-rot fungus Echinodontium taxodii 2538

A novel laccase was isolated and characterized from a new selective lignin-degrading white-rot fungus Echinodontium taxodii 2538, in which a high yield of laccase was obtained. No laccase isoenzyme was detected in the synthetic liquid media. The purified laccase (designated as EtL2538) had an apparent molecular mass of 56 kDa, pI value of 3.1, and N-terminal amino acid sequence of GIGPVTDLHIVNAAV. EtL2538 showed optimum pH at 3.0 and optimum temperature at 60 °C using ABTS as the substrate. EtL2538 revealed superior thermostability, and retained over 80% of its original activity after incubation for 2 h at 50 °C. The laccase gene, etl2538, was also cloned and sequenced. This gene encoded a mature laccase protein containing 499 amino acids (aa) preceded by a signal peptide of 21 aa, and the deduced protein sequence contained four copper-binding conserved domains of typical laccase protein. EtL2538 was further used in lignin oxidation and dye decolorization. Even without the existence of redox mediators, EtL2538 could cleave the methoxyl groups and β-O-4 ether linkages in lignin from bamboo, and significantly decolorize malachite green and RBBR. These novel properties of EtL2538 may render it as a potential biocatalyst for biotechnological and environmental applications.     

The Use of Fluorescent Target Arrays for Assessment of T Cell Responses In vivo

The ability to monitor T cell responses in vivo is important for the development of our understanding of the immune response and the design of immunotherapies. Here we describe the use of fluorescent target array (FTA) technology, which utilizes vital dyes such as carboxyfluorescein succinimidyl ester (CFSE), violet laser excitable dyes (CellTrace Violet: CTV) and red laser excitable dyes (Cell Proliferation Dye eFluor 670: CPD) to combinatorially label mouse lymphocytes into >250 discernable fluorescent cell clusters. Cell clusters within these FTAs can be pulsed with major histocompatibility (MHC) class-I and MHC class-II binding peptides and thereby act as target cells for CD8⁺ and CD4⁺ T cells, respectively. These FTA cells remain viable and fully functional, and can therefore be administered into mice to allow assessment of CD8⁺ T cell-mediated killing of FTA target cells and CD4⁺ T cell-meditated help of FTA B cell target cells in real time in vivo by flow cytometry. Since >250 target cells can be assessed at once, the technique allows the monitoring of T cell responses against several antigen epitopes at several concentrations and in multiple replicates. As such, the technique can measure T cell responses at both a quantitative (e.g. the cumulative magnitude of the response) and a qualitative (e.g. functional avidity and epitope-cross reactivity of the response) level. Herein, we describe how these FTAs are constructed and give an example of how they can be applied to assess T cell responses induced by a recombinant pox virus vaccine.     

活性X-3B红染料对作物生态毒理指标影响的比较研究

采用作物种子暴露实验 ,研究了小麦、白菜和水稻等 3种不同作物对活性X 3B红染料污染暴露的生态毒理学指标 .结果表明 ,小麦、白菜和水稻对活性X 3B红染料污染暴露的反应各异 ,具体表现为对3种作物的种子发芽抑制半效应浓度分别为 896 5、70 10和 75 14mg·L-1,对根伸长抑制的半效应浓度分别为 6 5 34、5 882和 45 70mg·L-1;这 3种作物在从种子萌发起至第 7天时去残余种子或种皮后的鲜生物量、干生物量均随活性X 3B红暴露浓度的增加而降低 ,但鲜生物量的降低率普遍高于干生物量的降低率 ;当作物种子暴露于高浓度 (4 5 0 0和 5 0 0 0mg·L-1)的活性X 3B红时 ,小麦生物量受该有机染料污染暴露的影响最为敏感 ,水稻其次 ,白菜最为迟钝 ;就小麦、白菜和水稻等 3种作物对活性X 3B红染料污染暴露的抗性而言 ,小麦最强 ,白菜次之 ,水稻最弱 .     

不溶性着色蔗渣木聚糖检测木聚糖酶活性*

蔗渣木聚糖经1.4-丁二醇二环氧甘油醚与染料气巴蓝3GA交联形成不溶性着色固体。这种着色蔗渣木聚糖在pH4-8范围内表现稳定,用于评价木聚糖酶活性及在平板上检测产酶微生物具有简捷而灵敏的效果。     

Influence of NaCl and Na2SO4 on the kinetics and dye decolorization ability of crude laccase from Ganoderma lucidum

In a solid state medium using yellow passion fruit waste as substrate, the basidiomycete Ganoderma lucidum produced a laccase as the main ligninolytic enzyme. This crude enzyme presented Michaelian behavior with both substrates tested, namely 3-ethylbenzthiazoline-6-sulphonic acid (ABTS) and the anthraquinone dye remazol brilliant blue R (RBBR). The K_M’s for these substrates were, respectively, 0.232 × 10⁻³ and 0.602 × 10⁻³ M. The actions of NaCl and Na₂SO₄, two important salts usually found in textile wastewaters, were investigated. The enzyme was inhibited by NaCl, but not by Na₂SO₄. Inhibition by NaCl was of the mixed type with two different inhibition constants. The enzyme was able to completely decolorize RBBR in the presence of 1.0 M Na₂SO₄ and 50% decolorization was found in the presence of 0.1 M NaCl. Such properties certainly make the enzyme a good agent for textile dye effluent treatment considering the fact that wastewaters of this industry usually contain high concentrations of NaCl and Na₂SO₄.     

A single nucleotide polymorphism melt curve assay employing an intercalating dye probe fluorescence resonance energy transfer for forensic analysis

The characterization and use of DNA sequence polymorphisms are an important aspect of forensic analysis. A number of approaches are being explored for single nucleotide polymorphism (SNP) genotyping, but current detection methods are subject to limitations that adversely impact their utility for forensic analysis. We have developed a novel method for genotyping both single and multiple SNPs that uses an intercalating dye and a probe labeled with a single fluorophore to affect a fluorescence energy transfer. Melting curve analysis is then used to distinguish true alleles from mismatched alleles. We term the new method dye probe fluorescence resonance energy transfer (dpFRET). In the current work, development proceeded at first with synthetic DNA template testing to establish proof of concept for the chemistry involved, followed by the design of polymerase chain reaction (PCR)-based genomic DNA assays to demonstrate potential forensic applications. The loci chosen for testing included both nuclear (MHC DRB) and mitochondrial DNA (cytochrome b) genes. A preliminary assessment of the sensitivity limits of the technology indicated that dpFRET was capable of accurately genotyping DNA from one single diploid cell equivalent. This technology could also potentially impact a wide range of nonforensic disciplines to aid in discovery, screening, and association of DNA sequence polymorphisms.