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Purification and characterization of a laccase from the edible wild mushroom Tricholoma mongolicum
- Source :
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY. 20:1069-1076
- Publication Year :
- 2010
- Publisher :
- Journal of Microbiology and Biotechnology, 2010.
-
Abstract
- A novel laccase from Tricholoma mongolicum was purified by using a procedure which entailed ion exchange chromatography on DEAE-cellulose, CM-cellulose, Q-Sepharose, and FPLC-gel filtration on Superdex 75. The purified enzyme was obtained with a specific activity of 1480 U/mg-protein and a final yield of 15%. It was found to be a monomeric protein with a molecular mass of 66 kDa as estimated by sodium dodecyl sulphate-polyacrylamide gel electrophoresis. Its N-terminal amino acid sequence was GIGPVADLYVGNRIL, similar to some but different other mushroom laccase. The optimum pH and temperature for the purified enzyme were pH 2 to pH 3 and 30 degrees C, respectively. It displayed a low K(m) toward 2,7-azinobis (3-ethylbenzothiazolone-6-sulfonic acid) diammonium salt (ABTS) and high K(cat)/K(m) values. The purified laccase oxidized a wide range of lignin-related phenols, but exerted maximal activity on ABTS. It was significantly inhibited by Hg(2+) ions, and remarkably stimulated by Cu(2+) ions. It inhibited HIV-1 reverse transcriptase and proliferation of hepatoma HepG2 cells and breast cancer MCF7 cells with an IC50 of 0.65 microM, 1.4 microM, and 4.2 microM, respectively, indicating that it is also an antipathogenic protein.
- Subjects :
- Sodium
Molecular Sequence Data
Ion chromatography
chemistry.chemical_element
Applied Microbiology and Biotechnology
chemistry.chemical_compound
Sequence Analysis, Protein
Humans
Amino Acid Sequence
Benzothiazoles
Polyacrylamide gel electrophoresis
Cell Proliferation
Laccase
Gel electrophoresis
ABTS
Chromatography
Molecular mass
biology
Tricholoma
Hep G2 Cells
General Medicine
biology.organism_classification
HIV Reverse Transcriptase
Molecular Weight
Kinetics
chemistry
Biochemistry
Electrophoresis, Polyacrylamide Gel
Sulfonic Acids
Biotechnology
Subjects
Details
- ISSN :
- 17388872 and 10177825
- Volume :
- 20
- Database :
- OpenAIRE
- Journal :
- JOURNAL OF MICROBIOLOGY AND BIOTECHNOLOGY
- Accession number :
- edsair.doi.dedup.....ed2723087f00d50f533c2065686057a7