SIAT Scientists Made Progresses on Saccharomyces cerevisiae’s Biological Components Standardization and Applications in Metabolic Engineering01 Jul 2021 Research Progress
Figure 1. Integration of multiple copies of genes using Wicket
The quantitative relationship and spatial distance between the substrates in each reaction of the metabolic pathway have a significant impact on metabolic flow and final output of the target product. Funded by this project, the researchers established a new method (AProSS) to increase the yield of specific natural products in yeast by rapidly assembling artificial scaffold proteins. Through the modular design of protein domains and the changes in the type, number and location of the domains, the researchers successfully constructed scaffold proteins with different characteristics to regulate both the direction of metabolic flow and the yield of violacein and deoxyvioletin respectively by 29 % and 63% (Figure 2).
Figure 2. Using artificial scaffold protein to increase the production of violacein.
How to quickly construct and characterize artificial cells is an important issue in synthetic biology. In this project, the researchers developed a high-throughput, semi-quantitative phenotyping method to evaluate the growth of synthetic yeast by choosing the test conditions and standardization of statistical analysis methods. At the same time, the research team also designed an efficient chromosome assembly strategy based on CRISPR/Cas9-mediated Gene Conversion, which greatly saved the time for assembly of large-segment DNA. These two methods can improve efficiency in the process of characterization and construction of synthetic strains, which greatly promotes the development of synthetic genomics related projects, and is expected to provide an artificial chassis for yeast-based metabolic engineering (Figure 3).
Figure 3. Using semi-quantitative phenotyping and fine assembly strategies to improve the efficiency of chromosome synthesis.
Email: lj.sun @siat.ac.cn
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Wicket: A Versatile Tool for the Integration and Optimization of Exogenous Pathways in Saccharomyces cerevisiae
Artificial Protein Scaffold System (AProSS): An efficient method to optimize exogenous metabolic pathways in Saccharomyces cerevisiae
Improving Chromosome Synthesis with a Semiquantitative Phenotypic Assay and Refined Assembly Strategy