Expression of CotA laccase in Pichia pastoris and its electrocatalytic sensing application for hydrogen peroxide
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| 008 | 210128e20151101xx s 000 0 eng | ||
| 024 | 7 | 0 | |a 10.1007/s00253-015-6720-0 |2 doi |
| 035 | |a (NATIONALLICENCE)springer-10.1007/s00253-015-6720-0 | ||
| 245 | 0 | 0 | |a Expression of CotA laccase in Pichia pastoris and its electrocatalytic sensing application for hydrogen peroxide |h [Elektronische Daten] |c [Lili Fan, Min Zhao, Yan Wang] |
| 520 | 3 | |a The CotA laccase from Bacillus subtilis WD23 was successfully overexpressed in Pichia pastoris, and the production level reached 891.2U/L. The recombinant CotA laccase was purified to homogeneity. The optimal enzymatic activity was found at pH 4.6, 6.6, and 6.8 for 2, 2′-azino-bis (3-ethylbenzothiazoline-6-sulfonate) (ABTS), 4-hydroxy-3, 5-dimethoxybenzaldehyde azine (SGZ), and 2, 6-dimethoxyphenol (2, 6-DMP) oxidation, respectively. The maximal enzyme activity was observed at 80°C with SGZ as a substrate. The kinetic constant K m values for ABTS, SGZ, and 2, 6-DMP were 162±20, 24±2, and 166±18μM, respectively, with corresponding k cat values of 15±1.0, 7.6±1.5, and 0.87±0.1s−1. Remarkably, the laccase activity increased to 561.9% of its initial activity at pH 9.0 after 7days of incubation and the half-life of laccase inactivation was approximately 3h at 80°C, which indicated that the recombinant CotA was a highly thermo-alkali-stable laccase. Bioelectrocatalytic reduction of H2O2 by the CotA laccase was detected when the recombinant CotA was adsorbed on pyrogenation graphite electrodes. Based on the bioelectrocatalytic reduction, a mediator-free amperometric biosensor for hydrogen peroxide was designed. The linear range of the H2O2 biosensor was from 0.05 to 4.75mM, with a detection limit of 3.1μM. The amperometric biosensor for H2O2 by CotA-modified electrode is a novel application for CotA laccase. | |
| 540 | |a Springer-Verlag Berlin Heidelberg, 2015 | ||
| 690 | 7 | |a CotA |2 nationallicence | |
| 690 | 7 | |a Laccase |2 nationallicence | |
| 690 | 7 | |a Secretory expression |2 nationallicence | |
| 690 | 7 | |a Bioelectrocatalytic |2 nationallicence | |
| 690 | 7 | |a H2O2 biosensor |2 nationallicence | |
| 700 | 1 | |a Fan |D Lili |u Academy of Fundamental and Interdisciplinary Sciences, Harbin Institute of Technology, No. 2, Yikuang Street, 150001, Harbin, China |4 aut | |
| 700 | 1 | |a Zhao |D Min |u College of Life Science, Northeast Forestry University, 150040, Harbin, China |4 aut | |
| 700 | 1 | |a Wang |D Yan |u Academy of Fundamental and Interdisciplinary Sciences, Harbin Institute of Technology, No. 2, Yikuang Street, 150001, Harbin, China |4 aut | |
| 773 | 0 | |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/22(2015-11-01), 9483-9493 |x 0175-7598 |q 99:22<9483 |1 2015 |2 99 |o 253 | |
| 856 | 4 | 0 | |u https://doi.org/10.1007/s00253-015-6720-0 |q text/html |z Onlinezugriff via DOI |
| 898 | |a BK010053 |b XK010053 |c XK010000 | ||
| 900 | 7 | |a Metadata rights reserved |b Springer special CC-BY-NC licence |2 nationallicence | |
| 908 | |D 1 |a research-article |2 jats | ||
| 949 | |B NATIONALLICENCE |F NATIONALLICENCE |b NL-springer | ||
| 950 | |B NATIONALLICENCE |P 856 |E 40 |u https://doi.org/10.1007/s00253-015-6720-0 |q text/html |z Onlinezugriff via DOI | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Fan |D Lili |u Academy of Fundamental and Interdisciplinary Sciences, Harbin Institute of Technology, No. 2, Yikuang Street, 150001, Harbin, China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Zhao |D Min |u College of Life Science, Northeast Forestry University, 150040, Harbin, China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 700 |E 1- |a Wang |D Yan |u Academy of Fundamental and Interdisciplinary Sciences, Harbin Institute of Technology, No. 2, Yikuang Street, 150001, Harbin, China |4 aut | ||
| 950 | |B NATIONALLICENCE |P 773 |E 0- |t Applied Microbiology and Biotechnology |d Springer Berlin Heidelberg |g 99/22(2015-11-01), 9483-9493 |x 0175-7598 |q 99:22<9483 |1 2015 |2 99 |o 253 | ||