Study on the structure-function correlation of CO_2-fixation enzymes toward the industrial application

CO_2固定酶的结构-功能相关性研究及工业应用

• 批准号: 05453122
• 负责人: KAI Yasushi
• 金额: $ 3.2万
• 依托单位: Osaka University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for General Scientific Research (B)
• 财政年份: 1993
• 资助国家: 日本
• 起止时间: 1993 至 1994
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-05453122/
• 关键词: X-ray structure analysis; CO_2-fixation enzymes; spinach; 2.0 A resolution; RuBisCo; phosphoenolpyrubicacidcarboxylase (PEPC); reflection-intensity-measurement; 二酸化炭素固定; リブロースビスリン酸カルボキシラーゼ; 結晶構造解析

The spinach RuBisCO (ribulose-1,5-bisphosphate carboxylase oxygenase) were purified and crystallized by the use of polyethylene glycol as a salt for salting out during the purification process and a precipitant for crystallization. The X-ray diffraction data were measured by the use of synchrotron radiation in Tsukuba KEK.The refinement of the crystal structure was proceeded up to the current R value of 19.0% for 2.0 ? 81 resolution data. The comparison of the molecular structure obtained by the present work with the spinach RuBisCO obtained by use of ammonium sulfate as a salt and precipitant are under investigation. The difference in the enzymatic function between these RuBisCO is expected to understand from the structure-function correlation based on the high resolution molecular structures.

菠菜RuBisCO（核酮糖-1，5-二磷酸羧化酶加氧酶）通过使用聚乙二醇作为在纯化过程中用于盐析的盐和用于结晶的沉淀剂来纯化和结晶。X射线衍射数据的测量使用同步辐射在筑波KEK的。晶体结构的细化进行到目前的R值为19.0%，为2.0？81分辨率数据。通过本工作获得的分子结构与菠菜RuBisCO通过使用硫酸铵作为盐和沉淀剂获得的比较正在调查中。这些RuBisCO之间的酶功能的差异有望从基于高分辨率分子结构的结构-功能相关性来理解。

项目成果

井上 豪: "Refined Crystal Structure of Pseudoazurin from Methylobacterium extorauens AML at 1.5A Resolution" Acta Crystallography. D50(3). 317-328 (1994)

Go Inoue：“1.5A 分辨率下来自甲基杆菌 AML 的假天青蛋白的精制晶体结构”《晶体学报》D50(3) (1994)。

原田 繁春: "Crystallization and Preliminary X-ray Studies of a protease from Pseudomonas aeruginosa" Journal of Molecular Biology. 230. 1315-1316 (1993)

Shigeharu Harada：“铜绿假单胞菌蛋白酶的结晶和初步 X 射线研究”《分子生物学杂志》230。1315-1316 (1993)

T.Inoue, N.Shibata, H.Nakanishi, S.Koyama, H.Ishii, Y.Kai, S.Harada, N.Kasai, Y.Ohshiro, S.Suzuki, T.Kohzuma, K.Yamaguchi, S.Shidara and H.Iwasaki: "Structure of Azurin from Achromobacter xylosoxidans NCIB11015 at 2.5? 81 Resolution" J.Biochem.116. 1193-1

T.Inoue、N.Shibata、H.Nakanishi、S.Koyama、H.Ishii、Y.Kai、S.Harada、N.Kasai、Y.Ohshiro、S.Suzuki、T.Kohzuma、K.Yamaguchi、S.

井上 豪: "Refined Crystal Structure of Pseudoazurin from Methylobacterium extorquens AMI at 1.5A Resolution" Acta Crystallography. D50(3). 317-328 (1994)

Go Inoue：“1.5A 分辨率下来自扭动甲基杆菌 AMI 的假天青蛋白的精制晶体结构”《晶体学报》D50(3) (1994)。

原田 繁春: "Crystallization and Preliminary X-ray Diffraction Studies of Curculinn ANew Type of Sweet Protein Having Taste-modifyeng Action" Journal of Molecular Bio. 238. 286-287 (1994)

Shigeharu Harada：“仙茅素的结晶和初步 X 射线衍射研究，一种具有味道修饰作用的新型甜味蛋白”，《分子生物杂志》238. 286-287 (1994)。