Laboratory of Natural Medicine and Molecular Engineering, College of Plant Science and Technology, Huazhong Agricultural University, Wuhan 430070, China.
National & Local Joint Engineering Research Center for Medicinal Plant Breeding and Cultivation, Wuhan 430070, China.
ACS Synth Biol. 2021 Nov 19;10(11):2938-2946. doi: 10.1021/acssynbio.1c00175. Epub 2021 Nov 1.
The enzymes immobilized through yeast surface display (YSD) can be used in metabolic pathway reconstruction as alternatives to the enzymes isolated or purified through conventional biochemistry methods. They can be easily prepared by growing and collecting yeast cells harboring display constructs. This may provide an economical method for enriching certain enzymes for biochemistry characterization and application. Herein, we took the advantage of one-pot cascade reactions catalyzed by YSD-immobilized enzymes in the mevalonate pathway to produce geraniol . YSD-immobilized enzymes of 10 cascade reactions for geraniol production, together with optimization of catalytic components, cofactor regeneration, and byproduct removal, achieved a final yield of 7.55 mg L after seven cycles. This study demonstrated that it is feasible to reconstitute a complex multi-enzymatic system for the chemical biosynthesis by exploiting YSD-immobilized cascade enzymes.
通过酵母表面展示（YSD）固定化的酶可以用作代谢途径重建中的替代物，替代通过传统生物化学方法分离或纯化的酶。它们可以通过生长和收集携带展示构建体的酵母细胞来轻松制备。这可能为丰富某些酶以进行生物化学特性分析和应用提供了一种经济的方法。在此，我们利用 YSD 固定化酶在甲羟戊酸途径中催化的一锅级联反应来生产香叶醇。YSD 固定化酶用于香叶醇生产的 10 级级联反应，以及对催化成分、辅因子再生和副产物去除的优化，在经过 7 个循环后最终产率达到 7.55mg/L。本研究表明，通过利用 YSD 固定化级联酶，可以重新构建用于化学生物合成的复杂多酶系统。
