Structural and functional analysis of insoluble-polysaccharide-hydrolyzing-enzyme by chimeric enzymes construction.
通过嵌合酶构建对不溶性多糖水解酶进行结构和功能分析。
基本信息
- 批准号:07660093
- 负责人:
- 金额:$ 1.47万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (C)
- 财政年份:1995
- 资助国家:日本
- 起止时间:1995 至 1996
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
To understand the structure and function relationsip of the insoluble polysaccharide-hydrolyzing-enzymes, such as cellulase and xylanase, we planned the research that construct chimeric protein (enzyme) between cellulase of Aspergillus aculeatus and xylanase of Bacillus pumilus that based on both stereostructure of the enzymes. To produce the chimeric enzyme proteins, the genes of the both enzymes were used. The chemeric genes which named XCX and CXC,were constructed by using restriction site and/or PCR method. The constructed genes were identified by the nucleotide sequencing. Both genes were expressed in Escherichia coli. The chimeric proteins that expressed in E.coli were purified to homogeneity by conventional chromatographic tequnique. Both purified chimeric proteins were reacted to antiserum against cellulase and xylanase from original strains. The activity of the chimeric proteins were analyzed to sensitive substrates, which were 4-methylumbeiferyl-beta-D-cellobiose, cellotriose, and xylose, respectively. The activities of the chimeric proteins were less than ditectable level.
为了了解纤维素酶和木聚糖酶等难溶多糖水解酶的结构和功能关系,我们计划在这两种酶的立体结构的基础上构建针状曲霉的纤维素酶和短小芽孢杆菌的木聚糖酶之间的嵌合蛋白(酶)。为了产生嵌合酶蛋白,使用了这两种酶的基因。利用限制性内切酶切点和/或聚合酶链式反应的方法构建了XCX和CXC两个化学基因。通过核苷酸测序对构建的基因进行鉴定。这两个基因都在大肠杆菌中表达。用常规层析技术纯化了在大肠杆菌中表达的嵌合蛋白。两种纯化的嵌合蛋白均与原菌株的纤维素酶和木聚糖酶抗血清反应。在敏感底物4-亚甲基-β-D-纤维二糖、纤维三糖和木糖上分析嵌合蛋白的活性。嵌合蛋白的活性均低于可鉴定水平。
项目成果
期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
数据更新时间:{{ journalArticles.updateTime }}
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
数据更新时间:{{ journalArticles.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ monograph.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ sciAawards.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ conferencePapers.updateTime }}
{{ item.title }}
- 作者:
{{ item.author }}
数据更新时间:{{ patent.updateTime }}
OOI Toshihiko其他文献
OOI Toshihiko的其他文献
{{
item.title }}
{{ item.translation_title }}
- DOI:
{{ item.doi }} - 发表时间:
{{ item.publish_year }} - 期刊:
- 影响因子:{{ item.factor }}
- 作者:
{{ item.authors }} - 通讯作者:
{{ item.author }}
{{ truncateString('OOI Toshihiko', 18)}}的其他基金
Development of microbial factory for bioplastic production adapting inedible biomass hydrolysates
开发采用非食用生物质水解产物生产生物塑料的微生物工厂
- 批准号:
23580452 - 财政年份:2011
- 资助金额:
$ 1.47万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
相似国自然基金
植物根际促生菌Bacillus pumilus WP8在“土壤-植物”系统中穿梭定殖机制研究
- 批准号:41501256
- 批准年份:2015
- 资助金额:20.0 万元
- 项目类别:青年科学基金项目
相似海外基金
Anti-MRSA activity of a Bacillus pumilus natural isolate
短小芽孢杆菌天然分离物的抗 MRSA 活性
- 批准号:
7571733 - 财政年份:2009
- 资助金额:
$ 1.47万 - 项目类别:
Anti-MRSA activity of a Bacillus pumilus natural isolate
短小芽孢杆菌天然分离物的抗 MRSA 活性
- 批准号:
7849907 - 财政年份:2009
- 资助金额:
$ 1.47万 - 项目类别:
Comparison of primary, secondary and tertiary structure of xylanase of Bacillus pumilus and cellulase of Aspergillus acleatus.
短小芽孢杆菌木聚糖酶和曲霉纤维素酶一级、二级和三级结构的比较。
- 批准号:
03453129 - 财政年份:1991
- 资助金额:
$ 1.47万 - 项目类别:
Grant-in-Aid for General Scientific Research (B)
Structural analysis of Bacillus pumilus xylanase
短小芽孢杆菌木聚糖酶的结构分析
- 批准号:
62560105 - 财政年份:1987
- 资助金额:
$ 1.47万 - 项目类别:
Grant-in-Aid for General Scientific Research (C)