Purification, Biochemical Characterization and Applications of Pleurotus ostreatus ARC280 Laccase
Abdelmageed M. Othman, Ali M. Elshafei, Mohamed M. Hassan, Bakry M. Haroun, Maysa A. Elsayed, Ayman A. Farrag
Microbiology Research Journal International · pp. 1418–1439 · Published 3 Aug 2014
10.9734/BMRJ/2014/11218Abstract
Aims: To purify, characterize, and apply the laccase produced by submerged fermentation using an edible mushroom Pleurotus ostreatus ARC280. Study Design: Laccase purification and characterization were designed using the most recent approaches and statistical studies of triplicate results values. Place and Duration of Study: Department of Microbial Chemistry, Genetic Engineering and Biotechnology Division, National Research Centre, Dokki, Cairo, Egypt, between May 2011 and January 2013. Methodology: P. ostreatus ARC280 laccase was purified using ammonium sulfate precipitation (40-80%), followed by gel filtration using Sephadex G100 column chromatography. The resulted pure laccase was analyzed on SDS-PAGE (12%). Laccase activity parameters such as temperature, pH, stability, metal ions and kinetic constants were studied. Laccase was applied to reduce four tumor cell lines growth and as antibacterial and antifungal agent. Results: P. ostreatus ARC280 laccase was purified using ammonium sulphate followed by Sephadex G-100 chromatographic column by about 148 purification fold with Mr of 85kDa. Optimum P. ostreatus ARC280 purified laccase activity was recorded at 50ºC and at pH 6.0, 3.0, 4.5 for Syringaldazine (SGZ), 2,2'-azinobis-(3-ethylbenzthiazoline-6-sulfonic) acid (ABTS) and 2, 6-dimethoxyphenol (DMP) as substrates, respectively. The purified enzyme was more stable in alkaline pH range and retained about 37.42, 73.51, 85.65, 87.7, 88.49, 93.65, 92.86 and 100.0 % of the initial activity after 5hrs of incubation at pH 3.0, 4.0, 5.0, 6.0, 7.0, 8.0, 9.0 and 10.0, respectively. Hg2+ caused complete inhibition at all tested concentrations; however Mn2+ (2.5x10-3M) caused laccase activation by about 190 and 330% after 1 and 24 hrs, respectively. Km and Vmax were calculated and found to be 0.074, 2.857 and 0.476 µM and 1.563, 2.500 and 2.632 µmol min-1 for SGZ, DMP and ABTS, respectively. The purified enzyme has the ability to reduce four tested cell lines growth in vitro with percentage reduction of 16.8, 23.4, 15.2 and 23.4% for HePG2, HCT116, A549 and MCF7, respectively. On the other hand, the enzyme was found to have antibacterial and antifungal activities against Escherichia coli and Candida albicans respectively. Conclusion: This enzyme seems to be a prospective enzyme for further biotechnological exploitation such as anticancer and antimicrobial activity applications.
References (52)
- 1 PROTEIN MEASUREMENT WITH THE FOLIN PHENOL REAGENT [DOI]
- 2 Cleavage of Structural Proteins during the Assembly of the Head of Bacteriophage T4 [DOI]
- 3 Multicopper Oxidases and Oxygenases [DOI]
- 4 [73] Spectrophotometric and turbidimetric methods for measuring proteins [DOI]
- 5 Fungal laccases – occurrence and properties [DOI]
- 6 The structure and function of fungal laccases [DOI]
- 7 Oxidation of non‐phenolic substrates [DOI]
- 8 Biodegradation of lignocellulosics: microbial, chemical, and enzymatic aspects of the fungal attack of lignin.
- 9 Laccases: A Useful Group of Oxidoreductive Enzymes [DOI]
- 10 Laccase: Microbial Sources, Production, Purification, and Potential Biotechnological Applications [DOI]
- 11 Screening of Hepatoprotective Plant Components using a HepG2 Cell Cytotoxicity Assay [DOI]
- 12 A Novel White Laccase from Pleurotus ostreatus [DOI]
- 13 Laccases and Their Applications: A Patent Review [DOI]
- 14 Laccase isoenzymes of Pleurotus eryngii: characterization, catalytic properties, and participation in activation of molecular oxygen and Mn2+ oxidation [DOI]
- 15 Medicinal mushrooms in supportive cancer therapies: an approach to anti-cancer effects and putative mechanisms of action [DOI]
- 16 Production, purification and characterization of mid-redox potential laccase from a newly isolated Trichoderma harzianum WL1 [DOI]
- 17 Purification and biochemical characterization of extracellular laccase from the ascomycete Paraconiothyrium variabile [DOI]
- 18 Purification and characterization of laccase from the white-rot fungus Daedalea quercina and decolorization of synthetic dyes by the enzyme [DOI]
- 19 Industrial and biotechnological applications of ligninolytic enzymes of the basidiomycota: a review [DOI]
- 20 Immobilization of laccase onto spacer-arm attached non-porous poly(GMA/EGDMA) beads: Application for textile dye degradation [DOI]
- 21 The White-Rot Fungus Pleurotus ostreatus Secretes Laccase Isozymes with Different Substrate Specificities [DOI]
- 22 Properties of crude laccase from Trametes versicolor produced by solid-substrate fermentation [DOI]
- 23 Fermentation optimization and characterization of the laccase from Pleurotus ostreatus strain 10969 [DOI]
- 24 Purification and characterization of laccase produced by a white rot fungus Pleurotus sajor-caju under submerged culture condition and its potential in decolorization of azo dyes [DOI]
- 25 Purification and characterization of a thermostable laccase from the ascomycetes Cladosporium cladosporioides and its applications [DOI]
- 26 Purification and characterization of an extracellular laccase from solid-state culture of Pleurotus ostreatus HP-1 [DOI]
- 27 Kinetic differences of purified laccases from six Pleurotus ostreatus strains [DOI]
- 28 Characterization of a Novel Laccase Produced by the Wood-Rotting Fungus Phellinus ribis [DOI]
- 29 Purification of a laccase from fruiting bodies of the mushroom Pleurotus eryngii [DOI]
- 30 Purification, characterization and application of laccase from Trametes versicolor for colour and phenolic removal of olive mill wastewater in the presence of 1- hydroxybenzotriazole [DOI]
- 31 The effects of a serine protease, Alcalase®, on the adhesives of barnacle cyprids (Balanus amphitrite) [DOI]
- 32 Kinetic characterization of purified laccase produced from Trametes versicolor IBL-04 in solid state bio-processing of corncobs [DOI]
- 33 Kinetics and Energetics of Azo Dye Decolorization by Pycnoporus sanguineus [DOI]
- 34 A laccase with antiproliferative activity against tumor cells from an edible mushroom, white common Agrocybe cylindracea [DOI]
- 35 Laccases of Pleurotus ostreatus observed at different phases of its growth in submerged fermentation: production of a novel laccase isoform [DOI]
- 36 Purification and characterization of a novel trimeric and thermotolerant laccase produced from the ascomycete Scytalidium thermophilum strain [DOI]
- 37 Antimicrobial activity of cotton fabrics containing immobilized enzymes [DOI]
- 38 Yellow laccase from the fungus Pleurotus ostreatus D1: Purification and characterization [DOI]
- 39 Optimization of Cultural and Nutritional Parameters for the Production of Laccase by Pleurotus ostreatus ARC280 [DOI]
- 40 One-copper laccase-related enzyme from Marasmius sp.: Purification, characterization and bleaching of textile dyes [DOI]
- 41 Purification and characterization of a novel laccase from the edible mushroom Hericium coralloides [DOI]
- 42 Laccase-induced C–N coupling of substituted p-hydroquinones with p-aminobenzoic acid in comparison with known chemical routes [DOI]
- 43 Purification and characterization of a laccase from the basidiomycete Funalia trogii (Berk.) isolated in Tanzania [DOI]
- 44 A Laccase with HIV-1 Reverse Transcriptase Inhibitory Activity from the Broth of Mycelial Culture of the MushroomLentinus tigrinus [DOI]
- 45 Purification and characterization of laccase from Trametes hirsuta Bm-2 and its contribution to dye and effluent decolorization [DOI]
- 46 Extracellular Laccase from Pleurotus ostreatus Strain EM-1: Thermal Stability and Response to Metal Ions [DOI]
- 47 ANALYSIS OF FUNGAL CULTURES ISOLATED FROM ANAMALAI HILLS FOR LACCASEENZYME PRODUCTION EFFECT ON DYE DECOLORIZATION, ANTIMICROBIALACTIVITY
- 48 Transformation of 7-(4-Hydroxyphenylacetamido)cephalosporanic Acid into a New Cephalosporin Antibiotic, 7 -[1-Oxaspiro(2.5)octa-6-oxo-4,7-diene-2-carboxamido]cephalosporanic Acid, by Laccase [DOI]
- 49 Purification and characterization of three laccase isozymes from the white rot fungus Trametes sp. HS-03 [DOI]
- 50 Study and development of a 'smart' wound dressing technology which can detect and inhibit/kill the colonisation of pathogenic bacteria
- 51 Microbial Laccases and their Applications: A Review [DOI]
- 52 Purification of a New Isoform of Laccase from a Marasmius quercophilus Strain Isolated from a Cork Oak Litter (Quercus suber L.) [DOI]
Cited by 50
Ali M. Elshafei, Abdelmageed M. Othman, Mohamed M. Hassan, Bakry M. Haroun, Maysa A. Elsayed, Ayman A. Farrag · Biotechnology Journal International · 2015
A. Elshafei, M. Elsayed, Gamil Ibrahim · Scientific Reports · 2025
Rajpal Choudhary, Aman Verma, Abhishek Sharma · Journal of Analytical and Applied Pyrolysis · 2025
Malak Alshammary, E. Kotb, I. Ababutain · Biology · 2025
Eko Fuji Ariyanto, Anastasya Kania Farahana, Gabriella Sachiko Jannesha Sudirman · Journal of Experimental Pharmacology · 2025
Kexing Yan, Lifeng Guo, Benfeng Zhang · Journal of Fungi · 2023
Ahmed A. Hamed, A. Abd-elaziz, Manal M E Ghanem · Folia Microbiologica (Prague) · 2023
K. Mohan, Durairaj Karthick Rajan, T. Muralisankar · Reviews in Aquaculture · 2022
H. Ahmad · Cellulose · 2021
Abdelmageed M. Othman, U. Wollenberger · International Journal of Biological Macromolecules · 2020
Showing 26 of 50 known citations — external sources report more than can currently be individually listed.
Related research
- Identification and Characterization of Antibiotic Resistant Bacteria Isolates from In-Patients at the Specialist Hospital, Sokoto, Nigeria — shares topic coverage
- Synthesis, Characterization and Biological Evaluation of Thiazolidinedione Substituted Derivatives — shares topic coverage
- Biochemical Characteristics of Immobilized Chitinase from Alternaria infectoria — shares topic coverage
- Silver Nanoparticles: Biosynthesis, Characterization and Application on Cotton Fabrics — shares topic coverage
- Characterization of Detergent-compatible Alkaline Protease from Bacillus agaradhaerens MTCC 9416 — shares topic coverage
Article metrics
Real usage data collected on this platform.
151
Page views
0
PDF downloads
47
Outbound clicks
50
Citations
Views over time
Views by country
Approximate, from request IP at view time — not citizenship or institution. Countries with fewer than 5 views are grouped as "Other".
Traffic sources
Referring site, by host.
Views and downloads exclude known bots/crawlers. Citations combines this platform's own DOI-resolved index with each external source's own reported total — see Cited by above for individually listed citing works. Last refreshed 0 seconds ago.