Improving the catalytic performance of a GH11 xylanase by rational protein engineering
文献类型：期刊论文
| 作者 | Cheng, Ya-Shan1,2; Chen, Chun-Chi3; Huang, Jian-Wen1,2; Ko, Tzu-Ping4; Huang, Zhiyong3; Guo, Rey-Ting3 |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 2015-11-01 |
| 卷号 | 99期号:22页码:9503-9510 |
| 关键词 | Glycoside hydrolase Structure-based design Mutagenesis Glycosylation Thermostability |
| 英文摘要 | XynCDBFV from Neocallimastix patriciarum is among the most effective xylanases and holds great potentials in a wide variety of industrial applications. In the present study, several active site residues were modified referring to the instrumental information of the complex structure of XynCDBFV and xylooligosaccharides. Among the 12 single active site mutants, W125F and F163W show increased activity comparing to the wild-type protein. The double mutant W125F/F163W was then generated which displayed nearly 20 % increase in the enzyme activity. Although W125F/F163W showed 5 A degrees C reduction in the optimal temperature, it still preserves similar thermostability and is more active than the wild-type enzyme at temperatures lower than 60 A degrees C. These properties make the double mutant a suitable candidate for commercial applications that involve lower operating temperatures. Furthermore, we investigated the effect of N-glycosylation on the thermostability of XynCDBFV when expressed in the yeast strain Pichia pastoris for industrial use. Two potential glycosylation sites (Asn-37 and Asn-88) were examined, and their roles in enzyme performance were validated. We found that the N-glycosylations of XynCDBFV are related to both catalytic activity and heat stability, with Asn-37 motif playing a dominant role. Collectively, the enzymatic properties of XynCDBFV were improved by molecular engineering, and the influences of N-glycosylations on the enzyme have been clearly elucidated herein. |
| WOS标题词 | Science & Technology ; Life Sciences & Biomedicine |
| 类目[WOS] | Biotechnology & Applied Microbiology |
| 研究领域[WOS] | Biotechnology & Applied Microbiology |
| 关键词[WOS] | FAMILY 11 XYLANASE ; DIRECTED EVOLUTION ; NEOCALLIMASTIX-PATRICIARUM ; CELLULASE ACTIVITY ; FUNGAL XYLANASE ; GLYCOSYLATION ; THERMOSTABILITY ; IMPROVEMENT ; DESIGN ; MUTAGENESIS |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000363951800016 |
| 源URL | [http://124.16.173.210/handle/834782/1557]  |
| 专题 | 天津工业生物技术研究所_结构生物学与蛋白酶学实验室 郭瑞庭_期刊论文 |
| 作者单位 | 1.Genozyme Biotechnol Inc, Taipei 106, Taiwan 2.AsiaPac Biotechnol Co Ltd, Dongguan 523808, Peoples R China 3.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Ind Enzymes Natl Engn Lab, Tianjin 300308, Peoples R China 4.Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan |
| 推荐引用方式 GB/T 7714 | Cheng, Ya-Shan,Chen, Chun-Chi,Huang, Jian-Wen,et al. Improving the catalytic performance of a GH11 xylanase by rational protein engineering[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2015,99(22):9503-9510. |
| APA | Cheng, Ya-Shan,Chen, Chun-Chi,Huang, Jian-Wen,Ko, Tzu-Ping,Huang, Zhiyong,&Guo, Rey-Ting.(2015).Improving the catalytic performance of a GH11 xylanase by rational protein engineering.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,99(22),9503-9510. |
| MLA | Cheng, Ya-Shan,et al."Improving the catalytic performance of a GH11 xylanase by rational protein engineering".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 99.22(2015):9503-9510. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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