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Cell surface engineering to endow cells with the ability of bioremediation and its application

赋予细胞生物修复能力的细胞表面工程及其应用

基本信息

批准号：
10450309
负责人：
TANAKA Atsuo
金额：
$ 9.09万
依托单位：
KYOTO UNIVERSITY
依托单位国家：
日本
项目类别：
Grant-in-Aid for Scientific Research (B)
财政年份：
1998
资助国家：
日本
起止时间：
1998 至 1999
项目状态：
已结题

来源：
https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-10450309/
关键词：
Cell surface engineering Molecular targeting Arming yeast Bioremediation Lipase Cellulase Cellulose Waste oil アンカリング GPI-アンカー酵母分子育種 α-アグルチニン

项目摘要

Novel yeast cells armed with biocatalysts - glucoamylase, α-amylase, CM-cellulase, β-glucosidase, and lipase - were constructed by a cell surface engineering system of yeast Saccharomyces cerevisiae. These surface-engineered yeast cells were termed "Arming yeasts". The gene encoding Rhizopus oryzae glucamylase with its secretion signal peptide was fused with the gene encoding the C-terminal half of yeast α-agglutinin. Glucoamylase was shown to be displayed on the cell surface of S. cerevisiae in its active form, anchored covalently to the cell wall. S. cerevisiae is unable to utilize starch, while the arming cells could grow on starch as the sole carbon source. For enhancement of the ability to directly ferment starchy materials by the arming yeast, a surface-engineered yeast cell displaying two amylolytic enzymes was constructed. The gene encoding R. oryzae glucoamylase with its own secretion signal peptide and a truncated fragment of the a-amylase gene from Bacillus stearothermophilu … More s with the prepro secretion signal sequence of the yeast α-factor, respectively, were fused with the gene encoding the C-terminal half of the yeast α-agglutinin. The arming cell co-displaying glucoamylase and α-amylase could grow faster on starch as the sole carbon source than the cell displaying only glucoamylase. Furthermore, a novel cellulose-utilizing yeast cell displaying cellulolytic enzymes in their active forms on the cell surface of S. cerevisiae was constructed by the cell surface engineering. An arming yeast codisplaying FI-carboxymethylcellulase(CM-cellulase), one of the endo-type cellulase, and β-glucosidase from Aspergillus aculeatus was endowed with the ability of celloligosaccharide assimilation, suggesting the possibility that the assimilation of cellulosic materials may be carried out by S. cerevisiae expressing heterologous cellulase genes on the cell surface. Furthermore, a yeast cell armed with R. oryzae lipase was also constructed. The technique will be applied to endow cells with the ability of bioremediation. Less

利用酿酒酵母细胞表面工程技术，构建了携带葡萄糖淀粉酶、α-淀粉酶、羧甲基纤维素酶、β-葡萄糖苷酶和脂肪酶等生物催化剂的新型酵母细胞。这些表面工程化的酵母细胞被称为"武装酵母"。将编码根霉葡糖淀粉酶及其分泌信号肽的基因与编码酵母α-凝集素C-末端的基因融合。葡萄糖淀粉酶显示在S.酿酒酵母以其活性形式，共价锚定到细胞壁。S.酿酒酵母不能利用淀粉，而武装细胞可以生长在淀粉作为唯一的碳源。为了提高武装酵母直接发酵淀粉类物质的能力，构建了具有两种淀粉分解酶的表面工程酵母细胞。编码R.嗜热脂肪芽孢杆菌α-淀粉酶基因截短片段及自身分泌信号肽的α-葡萄糖淀粉酶 ...更多信息分别与酵母α-因子前分泌信号肽序列和酵母α-凝集素C-末端基因进行融合。同时展示糖化酶和α-淀粉酶的武装细胞在以淀粉为唯一碳源时比仅展示糖化酶的武装细胞生长更快。此外，一种新的利用纤维素的酵母细胞展示纤维素分解酶在其活性形式的细胞表面的S。通过细胞表面工程技术构建了酿酒酵母的真核表达载体。将一株同时表达纤维素酶和β-葡萄糖苷酶的武装酵母菌（Aspergillusaculeatus）转化为纤维素酶，研究了纤维素酶和纤维素酶对武装酵母菌纤维寡糖的同化作用。酿酒酵母在细胞表面表达异源纤维素酶基因。此外，用R.还构建了脂肪酶。该技术将被应用于赋予细胞生物修复能力。少