• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文期刊>Biotechnology Progress >Enzyme encapsulation in permeabilized Saccharomyces cerevisiae cells
【24h】

Enzyme encapsulation in permeabilized Saccharomyces cerevisiae cells

机译：透化啤酒酵母细胞中的酶包封

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

The Saccharomyces cerevisiae cell wall provides a semipermeable barrier that can retain intracellular proteins but still permits small molecules to pass through. When S. cerevisiae cells expressing E. coli lacZ are treated with detergent to extract the cell membrane, beta-galactosidase activity in the permeabilized cells is approximately 40% of the activity of the protein in cell extract. However, the permeabilized cells can easily be collected and reused over 15 times without appreciable loss in activity. Cell wall composition and thickness can be modified using different cell strains for enzyme expression or by mutating genes involved in cell wall biosynthesis or degradation. The Sigma1278b strain cell wall is less permeable than the walls of BY4742 and W303 cells, and deleting EXG1, which encodes a 1,3-beta-glucanase, can further reduce permeability. A short Zymolyase treatment can increase cell wall permeability without rupturing the cells. Encapsulating multiple enzymes in permeabilized cells can offer kinetic advantages over the same enzymes in solution. Regeneration of ATP from AMP by adenylate kinase and pyruvate kinase encapsulated in the same cell proceeded more rapidly than regeneration using a cell extract. Combining permeabilized cells containing adenylate kinase with permeabilized cells containing pyruvate kinase can also regenerate ATP from AMP, but the kinetics of this reaction are slower than regeneration using cell extract or permeabilized cells expressing both enzymes.
机译：酿酒酵母细胞壁提供了一种半透性屏障，可以保留细胞内蛋白质，但仍允许小分子通过。用去污剂处理表达大肠杆菌lacZ的酿酒酵母细胞以提取细胞膜时，透化细胞中的β-半乳糖苷酶活性约为细胞提取物中蛋白质活性的40％。但是，通透性细胞可以轻松收集并重复使用15次以上，而不会明显丧失活性。可以使用不同的细胞株进行酶表达或通过突变参与细胞壁生物合成或降解的基因来改变细胞壁的组成和厚度。 Sigma1278b应变细胞壁的渗透性不如BY4742和W303细胞壁，删除编码1，3-β-葡聚糖酶的EXG1可以进一步降低渗透性。短时间的酶合酶处理可以增加细胞壁的通透性，而不会使细胞破裂。与溶液中的相同酶相比，将多种酶包裹在透化细胞中可以提供动力学优势。通过封装在同一细胞中的腺苷酸激酶和丙酮酸激酶从AMP再生ATP的过程比使用细胞提取物再生的过程更快。将含有腺苷酸激酶的透化细胞与含有丙酮酸激酶的透化细胞组合也可以从AMP再生ATP，但是该反应的动力学比使用细胞提取物或表达两种酶的透化细胞的再生慢。

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