Determination of phenolic acids using Trametes versicolor laccase.

Ege University, Faculty of Science, Biochemistry Department, 35100 Bornova-Izmir, Turkey.
Talanta (Impact Factor: 3.5). 03/2007; 71(1):312-7. DOI: 10.1016/j.talanta.2006.04.032
Source: PubMed

ABSTRACT Two biosensors based on Trametes versicolor laccase (TvL) were developed for the determination of phenolic compounds. Commercial oxygen electrode and ferrocene-modified screen-printed graphite electrodes were used for preparation of laccase biosensors. The systems were calibrated for three phenolic acids. Linearity was obtained in the concentration range 0.1-1.0muM caffeic acid, 0.05-0.2muM ferulic acid, 2.0-14.0muM syringic acid for laccase immobilised on a commercial oxygen electrode and 2.0-30.0muM caffeic acid, 2.0-10.0muM ferulic acid, 4.0-30.0muM syringic acid for laccase immobilised on ferrocene-modified screen-printed electrodes. Furthermore, optimal pH, temperature and thermal stability studies were performed with the commercial oxygen electrode. Both electrodes were used for determination of a class of phenolic acids, achieving a cheap and fast tool and an easy to be used procedure for screening real samples of human plasma.

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    ABSTRACT: In this work, the potential of several aqueous two-phase systems (ATPS) for laccase recovery was evaluated. For this purpose, different types of ATPS were prepared and the partition coefficient of pure commercial laccase was determined in each one, at 23 °C. A total of 21 ATPS were investigated: 12 composed by a polymer and a salt and 9 composed by two different polymers. For polymer–salt ATPS, different compositions of the same biphasic system were also studied. Laccase partition coefficients (K) obtained were used to conclude about ATPS effectiveness for this enzyme recovery. According to the results, it was found that UCON-phosphate salts (K ≤ 0.604), PEG-Li2SO4 (K = 2.081) and PES-dextran (K = 1.911) ATPS can be interesting options for laccase extraction, with laccase partitioning toward opposite directions. However, the most effective ATPS for laccase extraction was UCON-K2HPO4, presenting K values from 0.272 up to 0.306. UCON is a thermo-separating polymer which facilitates its recovery and reutilization. Additionally, for the first time, laccase stability in different ATPS was investigated by incubating the enzyme in each equilibrium phase during a week, at room temperature. The results obtained showed that UCON-sulfate salts, UCON-NaH2PO4 and UCON-KH2PO4 ATPS are not recommended for laccase recovery since a high loss of activity was observed: approximately 88% for sulphates and 80% for both dihydrogen phosphates. The best stability results were obtained with PEG-sulfate salts. For these ATPS, laccase stability remained similar or improved over time.
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    ABSTRACT: Typically, the use of heterogeneous enzyme catalysis is aimed at sustainability, reusability, or enhanced functionality of the biocatalyst and is achieved by immobilizing enzymes onto a support matrix or at a defined interface. Controlled enzyme immobilization is particularly important in bioelectrocatalysis because the catalyst must be effectively connected to a transducer to exploit its activity. This Review discusses what must be addressed for coupling biocatalysts to an electrode and the toolbox of methods that are available for achieving this outcome. As an illustration, we focus on the immobilization and stabilization of laccases at electronic interfaces. Historically, laccases have been used for the decolorization of dyes and for the synthesis of bio‐organic compounds; however, more recently, they have been applied to the fields of sensing and energy harvesting.1–3 There is an ever‐increasing focus on the development of new energy technologies, in which laccases find application (e.g., as cathodic catalysts in enzymatic fuel cells). Herein, we discuss the heterogeneous laccase biocatalysts that have been reported over the past 10–15 years and discuss why laccases continue to be biotechnologically relevant enzymes. Various methods for the immobilization of laccases are described, including the use of nanoscale supports and a range of encapsulation and cross‐linking chemistries. We consider the application of immobilized laccases to the food industry, in the synthesis of pharmaceuticals, and in environmental applications, specifically in cases in which stabilization through heterogenization of the enzyme is critical to the application. We also include a consideration of electrochemical biosensors and the specific incorporation of laccases on the surfaces of transducers.
    ChemCatChem 06/2013; 5(1). · 5.04 Impact Factor
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    ABSTRACT: Studies were carried on the decolorization of the textile dye reactive blue 19 (RB 19) by a novel isolate of Coprinus plicatilis (C. plicatilis) fungi. We describe an in vitro optimization process for decolorization and its behavior under different conditions of carbon and nitrogen sources, pH, temperature and substrate concentration. The optimal conditions for decolorization were obtained in media containing intermediate concentrations of ammonium oxalate and glucose (10 g/L) as nitrogen and carbon sources, respectively, at 26[degree sign]C and pH = 5.5. Maximum decolorization efficiency against RB 19 achieved in this study was around 99%. Ultra-violet and visible (UV-vis) spectrophotometric analyses, before and after decolorization, suggest that decolorization was due to biodegradation. This effect was associated with laccase enzyme displaying good tolerance to a wide range of pH values, salt concentrations and temperatures, suggesting a potential role for this organism in the remediation of real dye containing effluents. In conclusion, laccase activity in C. plicatilis was firstly described in this study.
    Journal of Environmental Health Science and Engineering 02/2014; 12(1):49. · 1.01 Impact Factor


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