Rational design of Escherichia coli for L-isoleucine production.

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.

journal_name

ACS Synth Biol

journal_title

ACS synthetic biology

authors

Park JH,Oh JE,Lee KH,Kim JY,Lee SY

doi

10.1021/sb300071a

subject

Has Abstract

pub_date

2012-11-16 00:00:00

pages

532-40

issue

11

issn

2161-5063

journal_volume

1

pub_type

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