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Cited 46 time in webofscience Cited 51 time in scopus
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Controlling Citrate Synthase Expression by CRISPR/Cas9 Genome Editing for n-Butanol Production in Escherichia coli

Authors
Heo, Min-JiJung, Hwi-MinUm, JaeyongLee, Sang-WooOh, Min-Kyu
Issue Date
Feb-2017
Publisher
AMER CHEMICAL SOC
Keywords
Escherichia coli; CRISPR/Cas9; genome editing; 5 '-untranslated region; n-butanol
Citation
ACS SYNTHETIC BIOLOGY, v.6, no.2, pp 182 - 189
Pages
8
Indexed
SCIE
SCOPUS
Journal Title
ACS SYNTHETIC BIOLOGY
Volume
6
Number
2
Start Page
182
End Page
189
URI
https://scholarworks.korea.ac.kr/kumedicine/handle/2020.sw.kumedicine/29484
DOI
10.1021/acssynbio.6b00134
ISSN
2161-5063
Abstract
Genome editing using CRISPR/Cas9 was successfully demonstrated in Esherichia coli to effectively produce n-butanol in a defined medium under microaerobic condition. The butanol synthetic pathway genes including those encoding oxygen-tolerant alcohol dehydrogenase were overexpressed in metabolically engineered E. coli, resulting in 0.82 g/L butanol production. To increase butanol production, carbon flux from acetyl-CoA to citric acid cycle should be redirected to acetoacetyl-CoA. For this purpose, the 5'-untranslated region sequence of gltA encoding citrate synthase was designed using an expression prediction program, UTR designer, and modified using the CRISPR/Cas9 genome editing method to reduce its expression level. E. coli strains with decreased citrate synthase expression produced more butanol and the citrate synthase activity was correlated with butanol production. These results demonstrate that redistributing carbon flux using genome editing is an efficient engineering tool for metabolite overproduction.
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Lee, Sang Woo
Ansan Hospital (Department of Gastroenterology and Hepatology, Ansan Hospital)
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