Light-Induced Nanobody-Mediated Targeted Protein Degradation for Metabolic Flux Control.

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

ACS Synthetic Biology Pub Date : 2024-11-11 DOI:10.1021/acssynbio.4c00552

Allison Y Tang, Seyi Jung, César Carrasco-López, José L Avalos

引用次数: 0

Abstract

In metabolic engineering, increasing chemical production usually involves manipulating the expression levels of key enzymes. However, limited synthetic tools exist for modulating enzyme activity beyond the transcription level. Inspired by natural post-translational mechanisms, we present targeted enzyme degradation mediated by optically controlled nanobodies. We applied this method to a branched biosynthetic pathway, deoxyviolacein, and observed enhanced product specificity and yield. We then extend the biosynthesis pathway to violacein and show how simultaneous degradation of two target enzymes can further shift production profiles. Through the redirection of metabolic flux, we demonstrate how targeted enzyme degradation can be used to minimize unwanted intermediates and boost the formation of desired products.

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光诱导纳米抗体介导的靶向蛋白质降解用于代谢通量控制

在代谢工程中，提高化学品产量通常涉及操纵关键酶的表达水平。然而，在转录水平之外调节酶活性的合成工具非常有限。受自然翻译后机制的启发，我们提出了由光控纳米抗体介导的定向酶降解。我们将这种方法应用于脱氧紫草素这一分支生物合成途径，并观察到产品特异性和产量均有所提高。然后，我们将生物合成途径扩展到中草药苷，并展示了两种目标酶的同时降解如何进一步改变产量曲线。通过对代谢通量的重新定向，我们展示了如何利用目标酶降解来尽量减少不需要的中间产物并促进所需产物的形成。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

ACS Synthetic Biology 生物-

CiteScore

8.00

自引率

10.60%

发文量

380

审稿时长

6-12 weeks

期刊介绍： The journal is particularly interested in studies on the design and synthesis of new genetic circuits and gene products; computational methods in the design of systems; and integrative applied approaches to understanding disease and metabolism. Topics may include, but are not limited to: Design and optimization of genetic systems Genetic circuit design and their principles for their organization into programs Computational methods to aid the design of genetic systems Experimental methods to quantify genetic parts, circuits, and metabolic fluxes Genetic parts libraries: their creation, analysis, and ontological representation Protein engineering including computational design Metabolic engineering and cellular manufacturing, including biomass conversion Natural product access, engineering, and production Creative and innovative applications of cellular programming Medical applications, tissue engineering, and the programming of therapeutic cells Minimal cell design and construction Genomics and genome replacement strategies Viral engineering Automated and robotic assembly platforms for synthetic biology DNA synthesis methodologies Metagenomics and synthetic metagenomic analysis Bioinformatics applied to gene discovery, chemoinformatics, and pathway construction Gene optimization Methods for genome-scale measurements of transcription and metabolomics Systems biology and methods to integrate multiple data sources in vitro and cell-free synthetic biology and molecular programming Nucleic acid engineering.