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对葡萄糖浓度作出响应的细胞表面工程酵母的基因控制自聚集

Genetically controlled self-aggregation of cell-surface-engineered yeast responding to glucose concentration.

作者信息

Zou W, Ueda M, Tanaka A

机构信息

Laboratory of Applied Biological Chemistry, Department of Synthetic Chemistry and Biological Chemistry, Graduate School of Engineering, Kyoto University, Yoshida, Sakyo-ku, Kyoto 606-8501, Japan.

出版信息

Appl Environ Microbiol. 2001 May;67(5):2083-7. doi: 10.1128/AEM.67.5.2083-2087.2001.

DOI:10.1128/AEM.67.5.2083-2087.2001
PMID:11319085
原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC92840/
Abstract

We constructed an arming (cell-surface-engineered) yeast displaying two types of agglutinin (modified a-agglutinin and alpha-agglutinin) on the cell surface, with agglutination being independent of both mating type and pheromones. The modified a-agglutinin was artificially prepared by the fusion of the genes encoding Aga1p and Aga2p. The modified a-agglutinin could induce agglutination of cells displaying Agalpha1p (alpha-agglutinin). The upstream region of the isocitrate lyase gene of Candida tropicalis (UPR-ICL), active at a low glucose concentration, was used as the promoter to express the modified a-agglutinin- and alpha-agglutinin-encoding genes. The arming yeast displaying both agglutinins agglutinated and sedimented in response to decreased glucose concentration. When the glucose concentration was high, the arming yeast grew normally. In the late log phase, when the glucose concentration became very low, agglutination occurred suddenly and drastically and yeast cells sedimented completely. Sedimentation was confirmed by weighing the aggregated cells after filtration of the broth. Strains in which aggregation can be genetically controlled can be used in industrial processes in which the separation of yeast cells from the supernatant is necessary.

摘要

我们构建了一种武装(细胞表面工程化)酵母,其在细胞表面展示两种凝集素(修饰的a凝集素和α凝集素),凝集作用与交配型和信息素均无关。修饰的a凝集素是通过编码Aga1p和Aga2p的基因融合人工制备的。修饰的a凝集素可诱导展示Agalpha1p(α凝集素)的细胞发生凝集。热带假丝酵母异柠檬酸裂解酶基因的上游区域(UPR-ICL)在低葡萄糖浓度下具有活性,被用作启动子来表达修饰的a凝集素和α凝集素编码基因。展示两种凝集素的武装酵母会因葡萄糖浓度降低而发生凝集和沉淀。当葡萄糖浓度较高时,武装酵母正常生长。在对数后期,当葡萄糖浓度变得非常低时,凝集突然剧烈发生,酵母细胞完全沉淀。通过对肉汤过滤后聚集的细胞称重来确认沉淀。能够进行遗传控制凝集的菌株可用于需要从上清液中分离酵母细胞的工业过程。

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本文引用的文献

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AG alpha 1 is the structural gene for the Saccharomyces cerevisiae alpha-agglutinin, a cell surface glycoprotein involved in cell-cell interactions during mating.AG alpha 1是酿酒酵母α-凝集素的结构基因,α-凝集素是一种细胞表面糖蛋白,在交配过程中参与细胞间相互作用。
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