Assembly of Minicellulosomes on the Surface of Bacillus subtilis (Retracted article. See vol. 84, unsp. e02429-17, 2018)
Assembly of Minicellulosomes on the Surface of Bacillus subtilis (Retracted article. See vol. 84, unsp. e02429-17, 2018)
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DOI：
10.1128/aem.02599-10
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发表时间：
2011-07-01
影响因子：
4.4
通讯作者：
Clubb, Robert T.
中科院分区：
文献类型：
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作者：
Anderson, Timothy D.;Robson, Scott A.;Clubb, Robert T.
为了经济有效地生产生物燃料，需要新的方法将木质纤维素生物质转化为可发酵的糖。一种有前途的方法是使用纤维素体降解生物质，纤维素体是存在于纤维素分解梭菌和瘤胃球菌属物种中的表面展示的含有多纤维素酶的复合物。在这项研究中，我们创建了枯草芽孢杆菌的纤维素分解菌株，其展示一种或多种纤维素酶。含有适当细胞壁分选信号的蛋白质通过与炭疽芽孢杆菌分选酶A（SrtA）转肽酶共表达而共价锚定至肽聚糖。该方法用于将来自热纤梭菌的Cel 8A内切葡聚糖酶共价连接至细胞壁。此外，Cel 8A-锚定蛋白融合蛋白锚定在B的表面上。枯草杆菌通过与细胞壁附着的粘附素模块的非共价相互作用。我们还表明，它是可能的组装多酶复合物的细胞表面上。在细胞表面上展示了含有三种酶的微纤维素体;它由与大肠杆菌中产生的三种纤维素酶-锚定蛋白融合蛋白非共价结合的细胞壁附着的支架蛋白组成。B。枯草芽孢杆菌具有强大的遗传系统，目前广泛用于工业过程。因此，将更大、更精细的微纤维素体接枝到B的表面上。枯草芽孢杆菌可产生具有增加的效力的纤维素分解细菌，其可用于降解生物质。
To cost-efficiently produce biofuels, new methods are needed to convert lignocellulosic biomass into fermentable sugars. One promising approach is to degrade biomass using cellulosomes, which are surface-displayed multicellulase-containing complexes present in cellulolytic Clostridium and Ruminococcus species. In this study we created cellulolytic strains of Bacillus subtilis that display one or more cellulase enzymes. Proteins containing the appropriate cell wall sorting signal are covalently anchored to the peptidoglycan by coexpressing them with the Bacillus anthracis sortase A (SrtA) transpeptidase. This approach was used to covalently attach the Cel8A endoglucanase from Clostridium thermocellum to the cell wall. In addition, a Cel8A-dockerin fusion protein was anchored on the surface of B. subtilis via noncovalent interactions with a cell wall-attached cohesin module. We also demonstrate that it is possible to assemble multienzyme complexes on the cell surface. A three-enzyme-containing minicellulosome was displayed on the cell surface; it consisted of a cell wall-attached scaffoldin protein noncovalently bound to three cellulase-dockerin fusion proteins that were produced in Escherichia coli. B. subtilis has a robust genetic system and is currently used in a wide range of industrial processes. Thus, grafting larger, more elaborate minicellulosomes onto the surface of B. subtilis may yield cellulolytic bacteria with increased potency that can be used to degrade biomass.