Site-Directed Mutagenesis of Multicopper Oxidase from Hyperthermophilic Archaea for High-Voltage Biofuel Cells

被引：5

作者：

Takamura, Eiichiro ^{[1
]}

Taki, Shunsuke ^{[1
]}

Sakamoto, Hiroaki ^{[1
]}

Satomura, Takenori ^{[2
]}

Sakuraba, Haruhiko ^{[3
]}

Ohshima, Toshihisa ^{[4
]}

Suye, Shin-ichiro ^{[1
,2
]}

机构：

[1] Univ Fukui, Grad Sch Engn, Dept Frontier Fiber Technol & Sci, 3-9-1 Bunkyo, Fukui, Fukui, Japan

[2] Univ Fukui, Grad Sch Engn, Dept Appl Chem & Biotechnol, 3-9-1 Bunkyo, Fukui, Fukui, Japan

[3] Kagawa Univ, Fac Agr, Dept Appl Biol Sci, 2393 Ikenobe, Miki, Kagawa, Japan

[4] Osaka Inst Technol, Fac Engn, Dept Biomed Engn, 5-16-1 Asahi Ku, Osaka, Japan

来源：

APPLIED BIOCHEMISTRY AND BIOTECHNOLOGY | 2021年 / 193卷 / 02期

基金：

日本学术振兴会;

关键词：

Site-directed mutagenesis; Multicopper oxidase; Redox potential; Biocathode; Biofuel cell; PYROBACULUM-AEROPHILUM; LACCASE; CUEO; CONSTRUCTION; ELECTRODES; MUTATIONS; SURFACE; DESIGN;

D O I：

10.1007/s12010-020-03440-5

中图分类号：

Q5 [生物化学]; Q7 [分子生物学];

学科分类号：

071010 ; 081704 ;

摘要：

Enzymes from hyperthermophilic archaea are potential candidates for industrial use because of their superior pH, thermal, and long-term stability, and are expected to improve the long-term stability of biofuel cells (BFCs). However, the reported multicopper oxidase (MCO) from hyperthermophilic archaea has lower redox potential than MCOs from other organisms, which leads to a decrease in the cell voltage of BFCs. In this study, we attempted to positively shift the redox potential of the MCO from hyperthermophilic archaeonPyrobaculum aerophilum(McoP). Mutations (M470L and M470F) were introduced into the axial ligand of the T1 copper atom of McoP, and the enzymatic chemistry and redox potentials were compared with that of the parent (M470). The redox potentials of M470L and M470F shifted positively by about 0.07 V compared with that of M470. In addition, the catalytic activity of the mutants towards 2,2 '-azino-bis(3-ethylbenzothiazoline-6-sulphonic acid) (ABTS) increased 1.2-1.3-fold. The thermal stability of the mutants and the electrocatalytic performance for O(2)reduction of M470F was slightly reduced compared with that of M470. This research provides useful enzymes for application as biocathode catalysts for high-voltage BFCs.

引用

页码：492 / 501

页数：10

共 23 条

[1] Construction of a biointerface on a carbon nanotube surface for efficient electron transfer [J].