Screening of thermophilic aldolases and application for the synthesis of sugar related materials

嗜热醛缩酶的筛选及其在糖相关材料合成中的应用

• 批准号: 20560730
• 负责人: SAKURABA Haruhiko
• 金额: $ 2.91万
• 依托单位: Kagawa University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (C)
• 财政年份: 2008
• 资助国家: 日本
• 起止时间: 2008 至 2010
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-20560730/
• 关键词: 結晶構造解析; 超好熱菌; アーキア; 好熱菌; アルドラーゼ; DERA; キメラ酵素; PCRスワッピング; eDNA; KDGA; Sulfolobus

The hyperthermophilic 2-deoxy-D-ribose-5-phosphate aldolase (DERA) gene was amplified from environmental DNA by PCR. These fragments were swapped with corresponding sequence of the known hyperthermophilic DERA gene and the expression vector for the chimera enzyme was constructed. The specific activity of the recombinant chimera DERA produced in Escherichia coli was two times higher than that of the wild type enzyme. In addition, we succeeded in determination of the crystal structure of hyperthermophilic 2-keto-3-deoxygluconate aldolases.

利用PCR技术从环境DNA中扩增了极端嗜热2-脱氧-D-核糖-5-磷酸醛缩酶（DERA）基因。将这些片段与已知的极端嗜热DERA基因的相应序列交换，并构建嵌合酶的表达载体。在大肠杆菌中产生的重组嵌合体DERA的比活性是野生型酶的2倍。此外，我们成功地确定了超嗜热2-酮-3-脱氧葡萄糖酸醛缩酶的晶体结构。

项目成果

Crystallization and preliminary X-ray analysis of a novel dye-linked L-proline dehydrogenase from the aerobic hyperthermophilic archaeon Aeropyrum pernix

来自需氧超嗜热古菌 Aeropyrum pernix 的新型染料连接 L-脯氨酸脱氢酶的结晶和初步 X 射线分析

• 发表时间: 2010
• 期刊: Acta.Cryst.F Vol.66
• 作者: T.Satomura;H.Sakuraba;Y.Hara;T.Ohshima
• 通讯作者: T.Ohshima

Structure of L-aspartate oxidase from the hyperthermophilic archaeon Sulfolobus tokodaii

超嗜热古菌 Sulfolobus tokodaii 的 L-天冬氨酸氧化酶的结构

• 发表时间: 2008
• 期刊: Biochim Biophys Acta 1784
• 作者: Sakuraba;H.;Yoneda;K.;Asai;I.;Tsuge;H.;Katunuma;N.;Ohshima;T
• 通讯作者: T

Crystal structure of UDP‐galactose 4‐epimerase‐like l‐threonine dehydrogenase belonging to the intermediate short‐chain dehydrogenase‐reductase superfamily

• DOI: 10.1111/j.1742-4658.2010.07916.x
• 发表时间: 2010-12
• 期刊: The FEBS Journal
• 作者: K. Yoneda;H. Sakuraba;Ikuo Muraoka;T. Oikawa;T. Ohshima

Characterization of a novel dye-linked l-proline dehydrogenase from an aerobic hyperthermophilic archaeon, Pyrobaculum calidifontis

• DOI: 10.1007/s00253-010-2914-7
• 发表时间: 2011-02
• 期刊: Applied Microbiology and Biotechnology
• 影响因子: 5
• 作者: T. Satomura;Xiaodong Zhang;Y. Hara;Katsumi Doi;H. Sakuraba;T. Ohshima

The first crystal structure of L-lysine 6-dehydrogenase as an NAD-dependent amine dehydrogenase

L-赖氨酸 6-脱氢酶作为 NAD 依赖性胺脱氢酶的第一个晶体结构

• 发表时间: 2010
• 期刊: J.Biol.Chem. Vol.285
• 作者: K.Yoneda;J.Fukuda;H.Sakuraba;T.Ohshima
• 通讯作者: T.Ohshima