Improvement of hydrogen peroxide stability of Pleurotus eryngii versatile ligninolytic peroxidase by rational protein engineering
文献类型：期刊论文
| 作者 | Bao, Xue1,2; Huang, Xuenian1 ; Lu, Xuefeng1; Li, Jian-Jun1
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| 刊名 | ENZYME AND MICROBIAL TECHNOLOGY
 |
| 出版日期 | 2014-01-10 |
| 卷号 | 54期号:2014页码:51-58 |
| 关键词 | Versatile peroxidase H2O2 stability Rational protein engineering |
| 英文摘要 | Peroxide tolerant versatile peroxidases are required for industrial applications. In this study, rational protein engineering was performed to improve the oxidative stability of Pleurotus eryngii versatile ligninolytic peroxidase. Residues which are easily oxidized such as methionine, and close to H2O2-binding pocket and heme were identified for site-directed mutagenesis. Enzyme activity and steady-state kinetics were affected to different extent by different mutations. They were investigated for H2O2 stability, among which mutants A79L, P141A, M247L, M265L, M247L/M265L, A77E/181L, A77E/A79S/I81L, A77S/A79L/I81L, A77E/A79S/I81L/M265L, A77E/A79S/I81L/M247L/M265L, and A77E/A79S/I81L/S168A showed significantly increased oxidative tolerance, proving that oxidizable residues such as Met247 and Met265, residues close to heme like Pro141 and H2O2-binding pocket such as Ala77, Ala79, and Ile81 exerted important impact on H2O2 stability. Double and triple mutants demonstrated some additive or synergistic effects, which were only inactivated by higher concentration H2O2, whereas multiple mutants A77E/A79S/I81L/M265L, A77E/A79S/I81L/M247L/M265L, and A77E/A79S/I81L/S168A did not. Importantly, mutants I81L, S168A, Met265, M247L/M265L, A77E/A79S/I81L, and A77E/A79S/I81L/M247L/M265L exhibited both improved catalytic efficiencies and H2O2 resistance. The enhanced oxidative stability could result from delayed or suppressed compound III formation and/or heme bleaching caused by replacement of some residues. The identified mutants with higher oxidative tolerance and catalytic efficiencies would be helpful for further improving the oxidative stability of versatile peroxidase. (C) 2013 Elsevier Inc. All rights reserved. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 学科主题 | 生物代谢工程 |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | SITE-DIRECTED MUTAGENESIS ; MANGANESE PEROXIDASE ; PHANEROCHAETE-CHRYSOSPORIUM ; HORSERADISH-PEROXIDASE ; ASCORBATE PEROXIDASE ; OXIDATION SITES ; H2O2 STABILITY ; EXPRESSION ; INACTIVATION ; EVOLUTION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000329531000009 |
| 公开日期 | 2014-03-21 |
| 源URL | [http://ir.qibebt.ac.cn:8080/handle/337004/1653]  |
| 专题 | 青岛生物能源与过程研究所_微生物代谢工程团队 |
| 作者单位 | 1.Chinese Acad Sci, Qingdao Inst Bioenergy & Bioproc Technol, Key Lab Biofuels, Qingdao 266101, Peoples R China 2.Univ Chinese Acad Sci, Beijing 100049, Peoples R China |
| 推荐引用方式 GB/T 7714 | Bao, Xue,Huang, Xuenian,Lu, Xuefeng,et al. Improvement of hydrogen peroxide stability of Pleurotus eryngii versatile ligninolytic peroxidase by rational protein engineering[J]. ENZYME AND MICROBIAL TECHNOLOGY,2014,54(2014):51-58. |
| APA | Bao, Xue,Huang, Xuenian,Lu, Xuefeng,&Li, Jian-Jun.(2014).Improvement of hydrogen peroxide stability of Pleurotus eryngii versatile ligninolytic peroxidase by rational protein engineering.ENZYME AND MICROBIAL TECHNOLOGY,54(2014),51-58. |
| MLA | Bao, Xue,et al."Improvement of hydrogen peroxide stability of Pleurotus eryngii versatile ligninolytic peroxidase by rational protein engineering".ENZYME AND MICROBIAL TECHNOLOGY 54.2014(2014):51-58. |
入库方式： OAI收割
来源：青岛生物能源与过程研究所
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