Studies on the Pycnoporus sanguineus CCT-4518 laccase purified by hydrophobic interaction chromatography

• Published in: Applied microbiology and biotechnology Vol. 75; no. 2; pp. 311 - 318
• Main Authors: Garcia, Telma Alves, Santiago, Mariângela Fontes, Ulhoa, Cirano José
• Format: Journal Article
• Language: English
• Published: Berlin Berlin/Heidelberg : Springer-Verlag 01.05.2007; Springer; Springer Nature B.V
• Subjects: Biological and medical sciences; Biotechnology; Catalytic oxidation; Characterization; Chromatography; Chromatography - methods; Culture Media; Enzyme Stability; Enzymes; Fundamental and applied biological sciences. Psychology; Hot Temperature; Hydrogen-Ion Concentration; Hydrophobic and Hydrophilic Interactions; Hydrophobic interaction; Kinetics; Laccase - chemistry; Laccase - isolation & purification; Laccase - metabolism; Life sciences; Molecular weight; Polyporaceae - enzymology; Polyporaceae - growth & development; Proteins; purification; Pycnoporus sanguineus; Sodium; Studies; Substrate Specificity; Temperature; Thermostable laccase; Hydrophobic interaction; Oxidoreductases; Enzyme; Hydrophobic chromatography; Laccase
• ISSN: 0175-7598; 1432-0614
• DOI: 10.1007/s00253-006-0817-4

A laccase from Pycnoporus sanguineus was purified by two steps using phenyl-Sepharose columm. A typical procedure provided 54.1-fold purification, with a yield of 8.37%, using syringaldazine as substrate. The molecular weight of the purified laccase was 69 and 68 kDa as estimated by 12% (w/v) SDS-PAGE gel and by gel filtration, respectively. The K m values for the substrates ABTS, syringaldazine, and guaiacol were 58, 8.3, and 370 μM, respectively. The enzyme's pH optimum for syringaldazine was 4.2 and optimal activity was 50°C. The enzyme showed to be thermostable because when kept at 50°C for 24 and 48 h it retained 93 and 76% activity. This laccase was inhibited by l-cysteine, β-mercaptoethanol, NaN₃, NaF, and HgCl₂.