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首页> 外文期刊>Applied Microbiology >Yeast Surface Display of Trifunctional Minicellulosomes for Simultaneous Saccharification and Fermentation of Cellulose to Ethanol
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Yeast Surface Display of Trifunctional Minicellulosomes for Simultaneous Saccharification and Fermentation of Cellulose to Ethanol

机译：酵母表面展示的三功能微型纤维素同时糖化和发酵成乙醇的酵母。

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By combining cellulase production, cellulose hydrolysis, and sugar fermentation into a single step, consolidated bioprocessing (CBP) represents a promising technology for biofuel production. Here we report engineering of Saccharomyces cerevisiae strains displaying a series of uni-, bi-, and trifunctional minicellulosomes. These minicellulosomes consist of (i) a miniscaffoldin containing a cellulose-binding domain and three cohesin modules, which was tethered to the cell surface through the yeast a-agglutinin adhesion receptor, and (ii) up to three types of cellulases, an endoglucanase, a cellobiohydrolase, and a β-glucosidase, each bearing a C-terminal dockerin. Cell surface assembly of the minicellulosomes was dependent on expression of the miniscaffoldin, indicating that formation of the complex was dictated by the high-affinity interactions between cohesins and dockerins. Compared to the unifunctional and bifunctional minicellulosomes, the quaternary trifunctional complexes showed enhanced enzyme-enzyme synergy and enzyme proximity synergy. More importantly, surface display of the trifunctional minicellulosomes gave yeast cells the ability to simultaneously break down and ferment phosphoric acid-swollen cellulose to ethanol with a titer of ～1.8 g/liter. To our knowledge, this is the first report of a recombinant yeast strain capable of producing cell-associated trifunctional minicellulosomes. The strain reported here represents a useful engineering platform for developing CBP-enabling microorganisms and elucidating principles of cellulosome construction and mode of action.
机译：通过将纤维素酶生产，纤维素水解和糖发酵整合为一个步骤，整合生物处理（CBP）代表了一种有前途的生物燃料生产技术。在这里，我们报告的酿酒酵母菌株的工程展示一系列的单，双和三功能微型纤维素体。这些微纤维素小体由（i）含有纤维素结合结构域和3个黏附素模块的微支架素组成，它们通过酵母α-凝集素粘附受体束缚在细胞表面，以及（ii）多达三种类型的纤维素酶，即一种内切葡聚糖酶，纤维二糖水解酶和β-葡萄糖苷酶，每个都带有一个C末端码头蛋白。微型细胞体的细胞表面组装取决于微型支架蛋白的表达，表明复合物的形成是由粘着蛋白和码头蛋白之间的高亲和力相互作用决定的。与单功能和双功能微纤维素体相比，季三功能复合物显示出增强的酶-酶协同作用和酶邻近协同作用。更重要的是，三功能微纤维素酶的表面展示使酵母细胞能够同时分解磷酸溶胀的纤维素并将其发酵为乙醇，滴度约为1.8 g / L。据我们所知，这是能够产生细胞相关的三功能微纤维素体的重组酵母菌株的首次报道。此处报道的菌株代表了一个有用的工程平台，可用于开发支持CBP的微生物并阐明纤维素小体构建原理和作用方式。

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