Dietary Change Enables Robust Growth-Coupling of Heterologous Methyltransferase Activity in Yeast,ACS Synthetic Biology

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Dietary Change Enables Robust Growth-Coupling of Heterologous Methyltransferase Activity in Yeast
ACS Synthetic Biology ( IF 3.7 ) Pub Date : 2020-11-12 , DOI: 10.1021/acssynbio.0c00348
Anne Sofie Lærke Hansen ₁ , Maitreya J. Dunham ₂ , Dushica Arsovska ₁ , Jie Zhang ₁ , Jay D. Keasling _{1,

3,

4,

5,

6} , Markus J. Herrgard _{1,

7} , Michael K. Jensen ₁

Affiliation

Genetic modifications of living organisms and proteins are made possible by a catalogue of molecular and synthetic biology tools, yet proper screening assays for genetic variants of interest continue to lag behind. Synthetic growth-coupling (GC) of enzyme activities offers a simple, inexpensive way to track such improvements. In this follow-up study we present the optimization of a recently established GC design for screening of heterologous methyltransferases (MTases) and related pathways in the yeast Saccharomyces cerevisiae. Specifically, upon testing different media compositions and genetic backgrounds, improved GC of different heterologous MTase activities is obtained. Furthermore, we demonstrate the strength of the system by screening a library of catechol O-MTase variants converting protocatechuic acid into vanillic acid. We demonstrated high correlation (R² = 0.775) between vanillic acid and cell density as a proxy for MTase activity. We envision that the improved MTase GC can aid evolution-guided optimization of biobased production processes for methylated compounds with yeast in the future.

中文翻译：

饮食变化使酵母中异源甲基转移酶活性得以强劲增长。

分子和合成生物学工具的目录使对活生物体和蛋白质的遗传修饰成为可能，但是针对感兴趣的遗传变异的适当筛选测定仍然滞后。酶活性的合成生长偶联（GC）提供了一种简单，廉价的方法来跟踪这种改善。在这项后续研究中，我们介绍了最近建立的用于筛选酵母酿酒酵母中异源甲基转移酶（MTase）及其相关途径的GC设计的优化方案。具体地，通过测试不同的培养基组成和遗传背景，获得了不同异源MTase活性的改进的GC。此外，我们通过筛选将原儿茶酸转化为香草酸的儿茶酚O-MTase变体文库来证明系统的强度。我们证明了香草酸和细胞密度之间的高度相关性（R ² = 0.775），可以作为MTase活性的代表。我们预想，改进的MTase GC可以在将来帮助基于进化的生物基生产工艺优化含酵母的甲基化化合物。

更新日期：2020-12-18

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