Modular Cloning Tools for Streptomyces spp. and Application to the De Novo Biosynthesis of Flavokermesic Acid.

IF 3.7 2区生物学 Q1 BIOCHEMICAL RESEARCH METHODS

ACS Synthetic Biology Pub Date : 2024-10-18 Epub Date: 2024-09-22 DOI:10.1021/acssynbio.4c00412

Jean-Malo Massicard, Delphine Noel, Andrea Calderari, André Le Jeune, Cyrille Pauthenier, Kira J Weissman

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Abstract

The filamentous Streptomyces are among the most prolific producers of bioactive natural products and are thus attractive chassis for the heterologous expression of native and designed biosynthetic pathways. Although suitable Streptomyces hosts exist, including genetically engineered cluster-free mutants, the approach is currently limited by the relative paucity of synthetic biology tools facilitating the de novo assembly of multicomponent gene clusters. Here, we report a modular system (MoClo) for Streptomyces including a set of adapted vectors and genetic elements, which allow for the construction of complete genetic circuits. Critical functional validation of each of the elements was obtained using the previously reported β-glucuronidase (GusA) reporter system. Furthermore, we provide proof-of-principle for the toolbox inS. albus, demonstrating the efficient assembly of a biosynthetic pathway to flavokermesic acid (FK), an advanced precursor of the commercially valuable carminic acid.

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链霉菌的模块化克隆工具及其在黄腐酸新生物合成中的应用。

丝状链霉菌是生物活性天然产物最丰富的生产者之一，因此是本地和设计生物合成途径异源表达的诱人底盘。虽然存在合适的链霉菌宿主，包括基因工程无簇突变体，但目前这种方法受限于促进从头组装多组分基因簇的合成生物学工具相对匮乏。在这里，我们报告了一种用于链霉菌的模块化系统（MoClo），其中包括一套适配载体和遗传元件，可以构建完整的遗传回路。利用之前报道的 β-葡糖醛酸酶（GusA）报告系统对每个元件进行了重要的功能验证。此外，我们还提供了工具箱在白僵菌（S. albus）中的原理验证，证明了黄腐酸（FK）生物合成途径的有效组装，黄腐酸是具有商业价值的卡米尼克酸的高级前体。

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.