• 主题
高级搜索  >
历史搜索 清空历史
首页> 外文会议>Microbial engineering >METHODOLOGY TO RAPIDLY ASSESS ENZYME CASCADES IN AID OF METABOLIC ENGINEERING OF HOST CELLS
【24h】

METHODOLOGY TO RAPIDLY ASSESS ENZYME CASCADES IN AID OF METABOLIC ENGINEERING OF HOST CELLS

机译：快速评估酶降解的方法论，以帮助宿主细胞的代谢工程

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

Chiral amino alcohols are compounds of pharmaceutical interest as they are building blocks of sphingolipids, antibiotics, and antiviral glycosidase inhibitors. Due to the challenges of chemical synthesis we recently developed two TK-TAm reaction cascades using natural and low cost feedstocks as substrates: a recycling cascade comprising of 2 enzymes and a sequential 3-step enzyme cascade yielding 30% and 1% conversion, respectively. In order to improve the conversion yield and aid the host strain engineering we used a combination of microscale experiments and statistical experimental design. For this we implemented a full factorial design to optimise pH, temperature and buffer type, followed by the implementation of Response Surface Methodology (RSM) for the optimisation of substrates and enzymes concentrations. We achieved 60% conversion for the recycling cascade and 3-fold improvement on the sequential pathway. Based on the results, limiting steps and individual requirements for host cell metabolic integration were identified expanding the understanding of the cascades without implementing extensive optimisation modelling. Therefore, the approach described here is ideal for exploratory work or when the interest is in defining the enzymatic expression levels required for microbial cell factories development.
机译：手性氨基醇是具有药用价值的化合物，因为它们是鞘脂，抗生素和抗病毒糖苷酶抑制剂的组成部分。由于化学合成的挑战，我们最近开发了两个使用天然和低成本原料作为底物的TK-TAm反应级联：包含2种酶的循环级联和依次产生30％和1％转化率的连续3步酶级联。为了提高转化率并协助宿主菌株工程设计，我们结合使用了微型实验和统计实验设计。为此，我们实施了全因子设计以优化pH，温度和缓冲液类型，然后实施了响应表面方法（RSM）以优化底物和酶的浓度。对于回收级联，我们实现了60％的转化率，在顺序途径上实现了3倍的改进。根据结果​​，确定了宿主细胞代谢整合的限制步骤和个人要求，从而扩大了对级联反应的理解，而无需实施广泛的优化建模。因此，此处描述的方法非常适合进行探索性工作或在确定微生物细胞工厂发展所需的酶表达水平时感兴趣。

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