The characterisation of immobilised lignin peroxidase by flow injection analysis |
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| Institution: | 1. Division of Pediatric Surgery, Ann & Robert H. Lurie Children’s Hospital, Northwestern University Feinberg School of Medicine;2. Surgical Outcomes and Quality Improvement Center, Department of Surgery, Northwestern University Feinberg School of Medicine;1. Department of Chemical and Biomolecular Engineering, University of Maryland, College Park, MD 20742, USA;2. Biophysics Program, University of Maryland, College Park, MD 20742, USA;1. Department of Pediatric Surgery, Beatrix Children''s Hospital, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands;2. Department of Pediatric Surgery, Pediatric Surgical Center Amsterdam, Amsterdam, the Netherlands;3. Department of Neonatology, Academic Medical Center, Amsterdam, the Netherlands;4. Department of Pediatric Radiology, Academic Medical Center, Amsterdam, the Netherlands;5. Department of Pediatric Radiology, Beatrix Children''s Hospital, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands;6. Department of Neonatology, VU University Medical Center, Amsterdam, the Netherlands;7. Department of Neonatology, Beatrix Children''s Hospital, University of Groningen, University Medical Center Groningen, Groningen, the Netherlands |
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| Abstract: | Immobilised lignin peroxidase has been investigated using a flow system in the steady state and by flow injection analysis (FIA). In the steady state, the extreme sensitivity of the enzyme towards inactivation by H2O2 resulted in a stable response only in the presence of saturating levels of organic substrate and at very low (10 μM) peroxide concentrations. By contrast, the low contact time during FIA led to a stable response to injections of 100 μM H2O2. At higher peroxide concentrations a reproducible inactivation was observed, allowing a study of factors affecting both activity and stability. Lignin peroxidase substrates that undergo at least semi-reversible oxidation/reduction, including high-molecular-weight lignin fractions, could be detected by electrochemical reduction of the oxidation products. With this detection system it was possible to demonstrate the role of veratryl alcohol as mediator. This mediated oxidation of lignin functioned only when all components were present simultaneously, and was not observed when lignin was separated from the site of veratryl alcohol oxidation. |
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