Biochemical characterization of an extremely stable pH-versatile laccase from Sporothrix carnis CPF-05.
| Autor: | Olajuyigbe FM; Enzyme Biotechnology and Environmental Health Unit, Department of Biochemistry, Federal University of Technology, Akure, Ondo State, Nigeria. Electronic address: folajuyi@futa.edu.ng., Fatokun CO; Enzyme Biotechnology and Environmental Health Unit, Department of Biochemistry, Federal University of Technology, Akure, Ondo State, Nigeria. |
|---|---|
| Jazyk: | angličtina |
| Zdroj: | International journal of biological macromolecules [Int J Biol Macromol] 2017 Jan; Vol. 94 (Pt A), pp. 535-543. Date of Electronic Publication: 2016 Oct 17. |
| DOI: | 10.1016/j.ijbiomac.2016.10.037 |
| Abstrakt: | Functionality of enzymes within narrow pH range and temperature is a major challenge which limits their industrial applications, hence, there is need to search for thermostable pH-versatile enzymes. Here, a novel thermostable pH-versatile laccase from Sporothrix carnis CPF-05 was purified by ion-exchange and gel filtration chromatography. Single protein band on sodium dodecyl sulfate polyacrylamide gel electrophoresis (SDS-PAGE) confirmed homogeneity of the enzyme with molecular weight of 56kDa. Enzyme yield was 3.9% and purification fold was 2.84. Purified laccase exhibited optimum activity at 50°C and retained 56% of its initial activity at 80°C after 180min of incubation with 2,2' azino-di-(3-ethylbenzothiazoline-6-sulfonic acid) (ABTS) as substrate. The enzyme had optimum pH of 7.0 and was stable over pH range of 3.0 to 11.0. Laccase activity was enhanced by Cu 2+ and Mn 2+ ions but inhibited by Ca 2+ , Mg 2+ , Ba 2+ and Hg 2+ ions. Purified laccase was mildly inhibited by urea, sodium azide and surfactants while exhibiting tolerance to organic solvents. The enzyme demonstrated broad substrate specificity. Kinetic parameters, K (Copyright © 2016 Elsevier B.V. All rights reserved.) |
| Databáze: | MEDLINE |
| Externí odkaz: |
|
Full Text from ScienceDirect