Rational design to improve thermostability and specific activity of the truncated Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase
文献类型：期刊论文
| 作者 | Huang, Jian-Wen1,2; Cheng, Ya-Shan3; Ko, Tzu-Ping5; Lin, Cheng-Yen1,2; Lai, Hui-Lin1,2; Chen, Chun-Chi6; Ma, Yanhe8; Zheng, Yingying8; Huang, Chun-Hsiang8; Zou, Peijian8 |
| 刊名 | APPLIED MICROBIOLOGY AND BIOTECHNOLOGY
 |
| 出版日期 | 2012-04-01 |
| 卷号 | 94期号:1页码:111-121 |
| 关键词 | Cellobiose Cellotetraose beta-Glucanase Crystal structure Synchrotron radiation |
| 英文摘要 | 1,3-1,4-beta-d-Glucanase has been widely used as a feed additive to help non-ruminant animals digest plant fibers, with potential in increasing nutrition turnover rate and reducing sanitary problems. Engineering of enzymes for better thermostability is of great importance because it not only can broaden their industrial applications, but also facilitate exploring the mechanism of enzyme stability from structural point of view. To obtain enzyme with higher thermostability and specific activity, structure-based rational design was carried out in this study. Eleven mutants of Fibrobacter succinogenes 1,3-1,4-beta-d-glucanase were constructed in attempt to improve the enzyme properties. In particular, the crude proteins expressed in Pichia pastoris were examined firstly to ensure that the protein productions meet the need for industrial fermentation. The crude protein of V18Y mutant showed a 2 A degrees C increment of Tm and W203Y showed similar to 30% increment of the specific activity. To further investigate the structure-function relationship, some mutants were expressed and purified from P. pastoris and Escherichia coli. Notably, the specific activity of purified W203Y which was expressed in E. coli was 63% higher than the wild-type protein. The double mutant V18Y/W203Y showed the same increments of Tm and specific activity as the single mutants did. When expressed and purified from E. coli, V18Y/W203Y showed similar pattern of thermostability increment and 75% higher specific activity. Furthermore, the apo-form and substrate complex structures of V18Y/W203Y were solved by X-ray crystallography. Analyzing protein structure of V18Y/W203Y helps elucidate how the mutations could enhance the protein stability and enzyme activity. |
| 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] | CRYSTAL-STRUCTURE ; PICHIA-PASTORIS ; CATALYTIC ROLES ; SITE RESIDUES ; ENZYMES ; GLYCOSYLATION ; PURIFICATION ; RECOGNITION ; MUTAGENESIS ; EXPRESSION |
| 收录类别 | SCI |
| 语种 | 英语 |
| WOS记录号 | WOS:000301747500011 |
| 源URL | [http://124.16.173.210/handle/834782/1305]  |
| 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.Natl Taiwan Univ, Inst Biotechnol, Taipei 106, Taiwan 4.Natl Taiwan Univ, Dept Anim Sci & Technol, Taipei 106, Taiwan 5.Acad Sinica, Inst Biol Chem, Taipei 115, Taiwan 6.Acad Sinica, Inst Biomed Sci, Taipei 115, Taiwan 7.Acad Sinica, Agr Biotechnol Res Ctr, Taipei 115, Taiwan 8.Chinese Acad Sci, Tianjin Inst Ind Biotechnol, Ind Enzymes Natl Engn Lab, Tianjin 300308, Peoples R China |
| 推荐引用方式 GB/T 7714 | Huang, Jian-Wen,Cheng, Ya-Shan,Ko, Tzu-Ping,et al. Rational design to improve thermostability and specific activity of the truncated Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase[J]. APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,2012,94(1):111-121. |
| APA | Huang, Jian-Wen.,Cheng, Ya-Shan.,Ko, Tzu-Ping.,Lin, Cheng-Yen.,Lai, Hui-Lin.,...&Guo, Rey-Ting.(2012).Rational design to improve thermostability and specific activity of the truncated Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase.APPLIED MICROBIOLOGY AND BIOTECHNOLOGY,94(1),111-121. |
| MLA | Huang, Jian-Wen,et al."Rational design to improve thermostability and specific activity of the truncated Fibrobacter succinogenes 1,3-1,4-beta-D-glucanase".APPLIED MICROBIOLOGY AND BIOTECHNOLOGY 94.1(2012):111-121. |
入库方式： OAI收割
来源：天津工业生物技术研究所
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