Purification,kinetics and spectral characterisation of a new versatile peroxidase from a Bjerkandera sp. isolate |
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| Affiliation: | 1. Departamento de Biotecnologia, Instituto National de Engenharia, Tecnologia e Inovação (INETI), Est. Paço do Lumiar 22, P-1649-038 Lisboa, Portugal;2. Escola Superior de Biotecnologia, Universidade Católica Portuguesa, R. Dr. António Bernardino de Almeida, P-4200-072 Porto, Portugal;3. Centre d’lngénierie des Protéines, Université de Liège, Institut de Chimie, B6 Sart Tilman, B-4000 Liège, Belgique;1. Department of Chemical Engineering, Applied Biocatalysis Unit Associated to IQAC (UAB-CSIC), School of Engineering, Universitat Autònoma de Barcelona, 08193 Bellaterra (Cerdanyola del Vallès), Catalunya, Spain;2. Centre of Chemistry, Institute of Chemistry, Technology and Metallurgy, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia;3. Department of Biochemistry, Faculty of Chemistry, University of Belgrade, Studentski trg 12-16, 11000 Belgrade, Serbia;1. Department of Chemical Sciences, University of Naples ‘Federico II’, via Cintia, 80126 Naples, Italy;2. Institute of Biostructures and Bioimaging, CNR, Via Mezzocannone 16, 80134 Naples, Italy;3. National Institute Biostructures and Biosystems, Inter-University Consortium, Viale Medaglie d’Oro 305, I-00136 Rome, Italy;1. The State Key Joint Laboratory of Environmental Simulation and Pollution Control, Department of Environment Science and Engineering, Tsinghua University, Beijing 100084, China;2. Department of Environment Science and Engineering, Kunming University of Science and Technology, Yunnan 650093, China;1. Departamento de Ingeniería Celular y Biocatálisis, Instituto de Biotecnología UNAM, Av. Universidad 2001 Chamilpa, 62210 Cuernavaca, Morelos, Mexico;2. Centro de Investigación en Biotecnología UAEM, Av. Universidad 2001 Chamilpa, 62210 Cuernavaca, Morelos, Mexico;1. Biofuels Institute, School of the Environment and Safety Engineering, Jiangsu University, Zhenjiang 212013, China;2. Botany Department, Faculty of Science, Tanta University, Tanta 31527, Egypt |
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| Abstract: | From the extracellular fluid of a novel strain of Bjerkandera sp., it was isolated, purified and identified the main enzyme responsible for Remazol Brilliant Blue R dye decolourisation. Such an enzyme is able to oxidise manganese, as well as veratryl alcohol and 2,6-dimethoxyphenol in manganese-independent reactions; hence, it can be included in the new group of versatile peroxidases. The molecular mass of said enzyme is ca. 45 kDa, and the N-terminal amino acid sequence obtained by Edman degradation is VAXPDGVNTA. The enzyme substrate range for oxidation of several phenolic and non-phenolic aromatic compounds was determined and the corresponding Michaelis–Menten kinetic constants calculated. Furthermore, spectrophotometric assays showing the Soret band and allowing observation of band shifts of the enzyme led to the conclusion that Bjerkandera strains may also synthesise at least two different versatile peroxidases, as happens with Pleurotus eryngii. |
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