Department of Chemical and Biomolecular Engineering
Korea Advanced Institute of Science and Technology

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[Cover Page]Rational Design of Escherichia coli for l-Isoleucine Production

Rational Design of Escherichia coli for l-Isoleucine Production

Jin Hwan Park , Jae Eun Oh , Kwang Ho Lee , Ji Young Kim , and Sang Yup Lee

 

ABSTRACT  

Metabolic engineering of Escherichia coli was performed to construct a 100% rationally engineered strain capable of overproducing l-isoleucine, an important branched-chain amino acid. The thrABC (encoding l-threonine biosynthetic enzymes), ilvA (encoding feedback-resistant threonine dehydratase), ilvIH (encoding feedback-resistant acetohydroxy acid synthase III), and ygaZH (encoding branched-chain amino acid exporter) genes were amplified by plasmid-based overexpression. The ilvCED (encoding l-isoleucine biosynthetic enzymes) and lrp (encoding global regulator Lrp) genes were also amplified by chromosomal promoter replacement in order to further increase the flux toward l-isoleucine. The final engineered E. coli strain was able to produce 9.46 g/L of l-isoleucine with a yield of 0.14 g/g of glucose by fed-batch culture. The overall design principles described here for the production of highly regulated product should be useful in designing strains for the production of other similar bioproducts.  

 

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등록자배성순

등록일2012-11-30

조회수9,852

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