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A Toolkit to Enable Hydrocarbon Conversion in Aqueous Environments
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高吞吐量吸收激活滴滴分类用于工程 化物脱酶

Ankit Jain1, Mariko Teshima2, Tomas Buryska1

  • 1Institute for Chemical and Bioengineering, Department of Chemistry & Applied Biosciences, ETH Zürich, Vladimir Prelog Weg 1, 8093, Zürich, Switzerland.

Angewandte Chemie (International ed. in English)
|August 1, 2024
PubMed
概括

这项研究引入了一种吸收活性滴滴分类器,用于高通量酶选,从而使生物催化剂的进化速度更快. 这种新方法成功地识别出一种改进的化脱酶变体,推进了可持续化学.

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科学领域:

  • 生物催化和酶工程 生物催化和酶工程
  • 微流体学 微流体学
  • 可持续化学 可持续化学

背景情况:

  • 生物催化剂的商业化正在增加,转向可持续方法.
  • 酶工程需要选大型突变库,这是传统方法的挑战.
  • 滴滴微流体提供高通量选,但受到光检测的限制.

研究的目的:

  • 开发一种新的吸收活性滴滴分类器,用于高通量酶选.
  • 扩大滴滴微流体的适用性,使其适用于更广泛的酶.
  • 为了能够对大型酶突变库进行稳健及时的选.

主要方法:

  • 在没有光学监控的情况下,开发了一种吸收活性滴滴分类器,用于千赫兹速率分类.
  • 使用NADH介导合试验对10^5个成员的化物脱酶库进行选.
  • 颜色测量检测 WST-1 formmazan产品以吸收率为基础的分类.

主要成果:

  • 成功演示了用于高通量选的吸收活性滴滴分类器.
  • 鉴定了一种具有51%的催化效率改善的阿尔代脱酶变体.
  • 工程变体在广泛的基质频谱中显示出整体活性增加.

结论:

  • 吸附活性滴滴分类器显著提高了酶进化能力.
  • 这项技术扩大了可以使用滴滴微流体学选的酶的范围.
  • 开发的系统有助于为可持续的化学过程发现改进的生物催化剂.