Construction of Lycopene Overproducing E. Coli Strains by Combining Systematic and Combinatorial Gene Knockout Targets  

Oleh:

Alper, Hal ; Miyaoku, Kohei ; Stephanopoulos, Gregory

Jenis: Article from Journal - ilmiah internasional
Dalam koleksi: Nature Biotechnology: The Science and Business of Biotechnology vol. 23 no. 5 (Mei 2005), page 612-616.
Topik: gene; lycopene; E. Coli; gene

Ketersediaan

Perpustakaan Pusat (Semanggi) Nomor Panggil: NN9.3 Non-tandon: 1 (dapat dipinjam: 0) Tandon: tidak ada

Lihat Detail Induk

Isi artikel

Identification of genes that affect the product accumulation phenotype of recombinant starings is an important problem in industrial strain construction and a central tenet of metabolic engineering. We have used systematic model-based) and combinatorial (transposon-based) methods to identify gene knockout targets that increase lycopene biosynthesis in strains of escherichia coli. We show that these two search strategies yield two distinct gene sets, which affect product synthesis either through an increase in precursor availability or through (largely unknown kinetic or regulatory mechanisms respectively. Exhaustic exploration of all possible combinations of the above gene sets yielded a unique set of 64 knockout strains spanning the metabolic landscape of systematic and combinatorial gene knockout targets. This included a global maximum strain exhibiting an 8,5 fold product increase over recombinant K12 wild type and a twofold increase over recombinant K12 wild type and a two fold increase over the engineered parental strain. These results were further validated in controlled culture conditions.

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