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Now showing items 1 - 8 of 8

  • Metabolic Engineering of Escherichia coli for Natural Product Biosynthesis

    Yang, Dongsoo   Park, Seon Young   Park, Yae Seul   Eun, Hyunmin   Lee, Sang Yup  

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  • Broad-Spectrum Gene Repression using Scaffold Engineering of Synthetic sRNAs

    Noh, Minho   Yoo, Seung Min   Yang, Dongsoo   Lee, Sang Yup  

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  • CRISPR/Cas9-coupled recombineering for metabolic engineering of Corynebacterium glutamicum

    Cho, Jae Sung   Choi, Kyeong Rok   Prabowo, Cindy Pricilia Surya   Shin, Jae Ho   Yang, Dongsoo   Jang, Jaedong   Lee, Sang Yup  

    Genome engineering of Corynebacterium glutamicum, an important industrial microorganism for amino acids production, currently relies on random mutagenesis and inefficient double crossover events. Here we report a rapid genome engineering strategy to scarlessly knock out one or more genes in C. glutamicum in sequential and iterative manner. Recombinase RecT is used to incorporate synthetic single-stranded oligodeoxyribonucleotides into the genome and CRISPR/Cas9 to counter-select negative mutants. We completed the system by engineering the respective plasmids harboring CRISPR/Cas9 and RecT for efficient curing such that multiple gene targets can be done iteratively and final strains will be free of plasmids. To demonstrate the system, seven different mutants were constructed within two weeks to study the combinatorial deletion effects of three different genes on the production of.-aminobutyric acid, an industrially relevant chemical of much interest. This genome engineering strategy will expedite metabolic engineering of C. glutamicum.
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  • Systems metabolic engineering as an enabling technology in accomplishing sustainable development goals

    Yang, Dongsoo   Cho, Jae Sung   Choi, Kyeong Rok   Kim, Hyun Uk   Lee, Sang Yup  

    With pressing issues arising in recent years, the United Nations proposed 17 Sustainable Development Goals (SDGs) as an agenda urging international cooperations for sustainable development. In this perspective, we examine the roles of systems metabolic engineering (SysME) and its contribution to improving the quality of life and protecting our environment, presenting how this field of study offers resolutions to the SDGs with relevant examples. We conclude with offering our opinion on the current state of SysME and the direction it should move forward in the generations to come, explicitly focusing on addressing the SDGs.
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  • Systems metabolic engineering as an enabling technology in accomplishing sustainable development goals

    Yang, Dongsoo   Cho, Jae Sung   Choi, Kyeong Rok   Kim, Hyun Uk   Lee, Sang Yup  

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  • Systems Metabolic Engineering Strategies: Integrating Systems and Synthetic Biology with Metabolic Engineering

    Choi, Kyeong Rok   Jang, Woo Dae   Yang, Dongsoo   Cho, Jae Sung   Park, Dahyeon   Lee, Sang Yup  

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  • Repurposing type III polyketide synthase as a malonyl-CoA biosensor for metabolic engineering in bacteria.

    Yang, Dongsoo   Kim, Won Jun   Yoo, Seung Min   Choi, Jong Hyun   Ha, Shin Hee   Lee, Mun Hee   Lee, Sang Yup  

    Malonyl-CoA is an important central metabolite for the production of diverse valuable chemicals including natural products, but its intracellular availability is often limited due to the competition with essential cellular metabolism. Several malonyl-CoA biosensors have been developed for high-throughput screening of targets increasing the malonyl-CoA pool. However, they are limited for use only in Escherichia coli and Saccharomyces cerevisiae and require multiple signal transduction steps. Here we report development of a colorimetric malonyl-CoA biosensor applicable in three industrially important bacteria: E. coli, Pseudomonas putida, and Corynebacterium glutamicum RppA, a type III polyketide synthase producing red-colored flaviolin, was repurposed as a malonyl-CoA biosensor in E. coli Strains with enhanced malonyl-CoA accumulation were identifiable by the colorimetric screening of cells showing increased red color. Other type III polyketide synthases could also be repurposed as malonyl-CoA biosensors. For target screening, a 1,858 synthetic small regulatory RNA library was constructed and applied to find 14 knockdown gene targets that generally enhanced malonyl-CoA level in E. coli These knockdown targets were applied to produce two polyketide (6-methylsalicylic acid and aloesone) and two phenylpropanoid (resveratrol and naringenin) compounds. Knocking down these genes alone or in combination, and also in multiple different E. coli strains for two polyketide cases, allowed rapid development of engineered strains capable of enhanced production of 6-methylsalicylic acid, aloesone, resveratrol, and naringenin to 440.3, 30.9, 51.8, and 103.8 mg/L, respectively. The malonyl-CoA biosensor developed here is a simple tool generally applicable to metabolic engineering of microorganisms to achieve enhanced production of malonyl-CoA-derived chemicals.=20
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  • Expanded synthetic small regulatory RNA expression platforms for rapid and multiplex gene expression knockdown

    Yang, Dongsoo   Yoo, Seung Min   Gu, Changdai   Ryu, Jae Yong   Lee, Jae Eun   Lee, Sang Yup  

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