...
  • 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Environmental Science & Technology >Cell Surface Display off Functional Macromolecule Fusions on Escherichia coli for Development of an Autofluorescent Whole-Cell Biocatalyst
【24h】

Cell Surface Display off Functional Macromolecule Fusions on Escherichia coli for Development of an Autofluorescent Whole-Cell Biocatalyst

机译：细胞表面展示功能性大分子融合在大肠杆菌上开发自发荧光的全细胞生物催化剂

获取原文
获取原文并翻译 | 示例
           
页面导航
  • 摘要
  • 著录项
  • 相似文献
  • 相关主题

摘要

At present Lpp-OmpA-mediated surface display has opened a new dimension in the development of whole-cell factories. Here, we report the surface display of methyl parathion hydrolase (MPH) and enhanced green fluorescent protein (EGFP) fusions (60 kDa) by employing the Lpp-OmpA chimera as an anchoring motif. A broad-host-range vector, pL0MG33, coding for Lpp-OmpA-MPH-GFP fusion protein was constructed for targeting the fusion protein onto the surface of Escherichia coli. The surface localization of fusion protein was demonstrated by Western blot analysis, immunofluorescence microscopy, and a protease accessibility experiment The surface-exposed fusion protein retains the MPH activity and GFP fluorescence. Anchorage of macromolecule fusions on the outer membrane neither inhibits cell growth nor affects cell viability, as shown by growth kinetics of cells and stability of resting cultures. The engineered ￡ coli with surface-expressed MPH-GFP has two major advantages over the same strain expressing cytosolic MPH-GFP, including 7-fold higher whole-cell activity and 2-fold stronger fluorescence. Moreover, the construct pL0MG33 can potentially be applied to various bacterial species for enhancing field use. This is the first report on the presentation of GFP fusions on the cell surface by Lpp-OmpA. Our results suggest that Lpp-OmpA is a useful tool for the functional display of macromolecule passenger proteins on the cell surface.
机译：目前，Lpp-OmpA介导的表面显示器为全细胞工厂的发展开辟了新的领域。在这里，我们报告通过使用Lpp-OmpA嵌合体作为锚定基序的甲基对硫磷水解酶（MPH）和增强的绿色荧光蛋白（EGFP）融合（60 kDa）的表面展示。构建用于编码Lpp-OmpA-MPH-GFP融合蛋白的广泛宿主载体pL0MG33，以将融合蛋白靶向大肠杆菌表面。通过蛋白质印迹分析，免疫荧光显微镜和蛋白酶可及性实验证明了融合蛋白的表面定位。表面暴露的融合蛋白保留了MPH活性和GFP荧光。如细胞生长动力学和静息培养物的稳定性所表明的那样，大分子融合物在外膜上的锚固既不抑制细胞生长也不影响细胞活力。与表达胞质MPH-GFP的同一菌株相比，具有表面表达的MPH-GFP的工程大肠杆菌具有两个主要优势，包括全细胞活性高7倍，荧光强度高2倍。此外，构建体pLOMG33可以潜在地应用于各种细菌种类以增强田间使用。这是有关Lpp-OmpA在细胞表面呈现GFP融合蛋白的第一份报告。我们的结果表明，Lpp-OmpA是在细胞表面上功能展示大分子过客蛋白的有用工具。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱：kefu@zhangqiaokeyan.com

京公网安备：11010802029741号 ICP备案号：京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有

  • 客服微信

  • 服务号