荧光假单胞菌染料脱色过氧化物酶DyP1B：表达、重组和与聚合木质素底物的反应。

4区生物学 Q3 Biochemistry, Genetics and Molecular Biology

Methods in enzymology Pub Date : 2025-01-01 Epub Date: 2025-02-09 DOI:10.1016/bs.mie.2025.01.049

Rahman Rahmanpour, Timothy D H Bugg

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引用次数: 0

摘要

已经鉴定出几种细菌染料脱色过氧化物酶，它们具有氧化木质素模型化合物和聚合物木质素的活性。本文介绍了从荧光假单胞菌Pf-5中提取木质素氧化过氧化物酶DyP1B的生化方法。本文介绍了：(1)酶纯化和血红素重组的方法；(2)稳态动力学酶分析；(3)预稳态动力学分析；(4)酶对高分子木质素底物的抗性测试。

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Dye-decolorising peroxidase DyP1B from Pseudomonas fluorescens: Expression, reconstitution and reaction with polymeric lignin substrates.

Several bacterial dye-decolorising peroxidases have been identified, that have activity for oxidation of lignin model compounds and polymeric lignin. This article describes biochemical methods for lignin-oxidising peroxidase DyP1B from Pseudomonas fluorescens Pf-5. The article presents methods for: (1) enzyme purification and heme reconstitution; (2) steady-state kinetic enzyme assays; (3) pre-steady state kinetic analysis; (4) testing of the enzyme against polymeric lignin substrates.

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来源期刊

Methods in enzymology 生物-生化研究方法

CiteScore

2.90

自引率

0.00%

发文量

308

审稿时长

3-6 weeks

期刊介绍： The critically acclaimed laboratory standard for almost 50 years, Methods in Enzymology is one of the most highly respected publications in the field of biochemistry. Each volume is eagerly awaited, frequently consulted, and praised by researchers and reviewers alike. Now with over 500 volumes the series contains much material still relevant today and is truly an essential publication for researchers in all fields of life sciences, including microbiology, biochemistry, cancer research and genetics-just to name a few. Five of the 2013 Nobel Laureates have edited or contributed to volumes of MIE.