Development of nanofabrication method of supermacromolecules on the yeast cell surface and its application to bioprocesses
酵母细胞表面超高分子纳米加工方法的发展及其在生物过程中的应用
基本信息
- 批准号:16360413
- 负责人:
- 金额:$ 9.41万
- 依托单位:
- 依托单位国家:日本
- 项目类别:Grant-in-Aid for Scientific Research (B)
- 财政年份:2004
- 资助国家:日本
- 起止时间:2004 至 2005
- 项目状态:已结题
- 来源:
- 关键词:
项目摘要
To reduce the cost of immobilized enzymes, heterogonous enzymes displaying recombinant microorganisms have been developed by many researchers. However, in a current cell-surface display system to perform the complicated reaction, it is necessary to control the ration of the proteins displayed on the cell surface. To overcome this problem, we constructed a novel cell-surface display system using two pairs of proteins that assemble each other, 1 pair namely ZZ domain of protein A derived from Staphylococcus aureus and Fc part of human immunoglobulin G. And 2 pair cohesin and dockerin of cellulosome from Clostridium cellulovorans. In this system, scaffolding protein (ZZ-Coh-Coh) is displayed on the cell surface and the target protein fused to the Fc or Dockerin are secreted. Using this system, recombinant Trichoderma reesei endoglucanase II was displayed on the yeast cell surface. Display of the EGII on the cell surface was confirmed by hydrolysis experiment using β-glucan as a substrate and EGII activity was detected in the cell-pellet fraction. Following experiment showed that, EGII and Aspergillus aculeatus β-glucosidase 1 were co-displayed on the cell surface. The resulting yeast cells hydrolyzed β-glucan to glucose. This study demonstrated simultaneous display of two enzymes on the yeast cell surface can be accomplished using this cell surface display system.
为了降低固定化酶的成本,许多研究人员开发了展示重组微生物的异源酶。然而,在当前的细胞表面展示系统中,为了进行复杂的反应,需要控制展示在细胞表面上的蛋白质的比例。为了克服这个问题,我们使用两对相互组装的蛋白质构建了一种新型细胞表面展示系统,其中一对即来自金黄色葡萄球菌的蛋白 A 的 ZZ 结构域和人免疫球蛋白 G 的 Fc 部分。以及两对来自纤维素梭菌的多纤维素酶体的粘连蛋白和坞蛋白。在该系统中,支架蛋白(ZZ-Coh-Coh)展示在细胞表面,并分泌与Fc或Dockerin融合的目标蛋白。使用该系统,重组里氏木霉内切葡聚糖酶 II 被展示在酵母细胞表面。通过使用β-葡聚糖作为底物的水解实验证实了EGII在细胞表面上的展示,并在细胞沉淀部分中检测到了EGII活性。后续实验表明,EGII与棘孢曲霉β-葡萄糖苷酶1共展示在细胞表面。所得酵母细胞将 β-葡聚糖水解为葡萄糖。这项研究证明,使用这种细胞表面展示系统可以在酵母细胞表面同时展示两种酶。
项目成果
期刊论文数量(3)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)
Ethanol fermentation from lignocellulosic hydrolysate by a recom binant xylose-and celloolingosaccharide-assimilationg veast strain.
通过重组木糖和纤维糖同化酵母菌株从木质纤维素水解产物中发酵乙醇。
- DOI:
- 发表时间:
- 期刊:
- 影响因子:0
- 作者:Katahira;S.;Mizuike;A.;Fukuda;H.;Kondo;A.
- 通讯作者:A.
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KONDO Akihiko其他文献
Butyryl-CoA:acetate CoA-transferase gene associated with the genus <i>Roseburia</i> is decreased in the gut microbiota of Japanese patients with ulcerative colitis
日本溃疡性结肠炎患者肠道菌群中与罗斯伯利亚属相关的丁酰辅酶A:乙酸辅酶A转移酶基因减少
- DOI:
10.12938/bmfh.18-029 - 发表时间:
2019 - 期刊:
- 影响因子:0
- 作者:
SHINOHARA Ryohei;SASAKI Kengo;INOUE Jun;HOSHI Namiko;FUKUDA Itsuko;SASAKI Daisuke;KONDO Akihiko;OSAWA Ro - 通讯作者:
OSAWA Ro
KONDO Akihiko的其他文献
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{{ truncateString('KONDO Akihiko', 18)}}的其他基金
A study on instability mechanism of granular materials by visualizing effective stress transmission in micro scale
通过微观尺度有效应力传递的可视化研究颗粒材料的失稳机制
- 批准号:
20K14830 - 财政年份:2020
- 资助金额:
$ 9.41万 - 项目类别:
Grant-in-Aid for Early-Career Scientists
Development of real-time visualization of stress distribution in granular material based on the mechanoluminescence phenomenon
基于力致发光现象的颗粒材料应力分布实时可视化的发展
- 批准号:
17K14725 - 财政年份:2017
- 资助金额:
$ 9.41万 - 项目类别:
Grant-in-Aid for Young Scientists (B)
evelopment of temperature responsible magnetic nanoparticles for targeting cancer cells
开发用于靶向癌细胞的温度响应磁性纳米颗粒
- 批准号:
23656528 - 财政年份:2011
- 资助金额:
$ 9.41万 - 项目类别:
Grant-in-Aid for Challenging Exploratory Research
Whole-cell biocatalyst with non-natural membrane structure
具有非天然膜结构的全细胞生物催化剂
- 批准号:
20360375 - 财政年份:2008
- 资助金额:
$ 9.41万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
Development of high efficiency cell-free protein production and purification system
高效无细胞蛋白生产和纯化系统的开发
- 批准号:
13650830 - 财政年份:2001
- 资助金额:
$ 9.41万 - 项目类别:
Grant-in-Aid for Scientific Research (C)
Development of bioprocess for production of clean biodiesel fuel from waste oil
开发利用废油生产清洁生物柴油燃料的生物工艺
- 批准号:
13555208 - 财政年份:2001
- 资助金额:
$ 9.41万 - 项目类别:
Grant-in-Aid for Scientific Research (B)
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