2 级微发酵

IF 3.7 Q2 BIOTECHNOLOGY & APPLIED MICROBIOLOGY
Shuai Li , Zhixia Ye , Eirik A. Moreb , Romel Menacho-Melgar , Maximillian Golovsky , Michael D. Lynch
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引用次数: 0

摘要

以细胞为基础的工厂可以生产多种产品。DNA 合成和基因组编辑技术的进步大大简化了这些工厂的设计和建造。生成数百甚至数千个用于评估的细胞工厂菌株变体变得前所未有的容易。这些进步更加需要标准化、高通量的方法来评估这些设计。为了实现这一目标,我们之前报道了工程大肠杆菌菌株的开发和相关的两阶段生产流程,以简化和标准化菌株工程、评估和放大。这种方法将生长(第 1 阶段)与生产(第 2 阶段)分离开来。磷酸盐耗竭是停止生长和诱导异源表达的触发器。在此,我们详细介绍了在两阶段微发酵中对工程大肠杆菌菌株进行评估的方案开发。这些方案可随时用于评估生产各种蛋白质和小分子产品的菌株。此外,通过详细介绍方案开发的方法,这些方法还可适用于其他细胞宿主,以及具有各种额外触发器的其他两阶段过程。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
2-Stage microfermentations

Cell based factories can be engineered to produce a wide variety of products. Advances in DNA synthesis and genome editing have greatly simplified the design and construction of these factories. It has never been easier to generate hundreds or even thousands of cell factory strain variants for evaluation. These advances have amplified the need for standardized, higher throughput means of evaluating these designs. Toward this goal, we have previously reported the development of engineered E. coli strains and associated 2-stage production processes to simplify and standardize strain engineering, evaluation and scale up. This approach relies on decoupling growth (stage 1), from production, which occurs in stationary phase (stage 2). Phosphate depletion is used as the trigger to stop growth as well as induce heterologous expression. Here, we describe in detail the development of protocols for the evaluation of engineered E. coli strains in 2-stage microfermentations. These protocols are readily adaptable to the evaluation of strains producing a wide variety of protein as well as small molecule products. Additionally, by detailing the approach to protocol development, these methods are also adaptable to additional cellular hosts, as well as other 2-stage processes with various additional triggers.

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来源期刊
Metabolic Engineering Communications
Metabolic Engineering Communications Medicine-Endocrinology, Diabetes and Metabolism
CiteScore
13.30
自引率
1.90%
发文量
22
审稿时长
18 weeks
期刊介绍: Metabolic Engineering Communications, a companion title to Metabolic Engineering (MBE), is devoted to publishing original research in the areas of metabolic engineering, synthetic biology, computational biology and systems biology for problems related to metabolism and the engineering of metabolism for the production of fuels, chemicals, and pharmaceuticals. The journal will carry articles on the design, construction, and analysis of biological systems ranging from pathway components to biological complexes and genomes (including genomic, analytical and bioinformatics methods) in suitable host cells to allow them to produce novel compounds of industrial and medical interest. Demonstrations of regulatory designs and synthetic circuits that alter the performance of biochemical pathways and cellular processes will also be presented. Metabolic Engineering Communications complements MBE by publishing articles that are either shorter than those published in the full journal, or which describe key elements of larger metabolic engineering efforts.
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