Abstract:

Using synthetic biology and metabolic engineering techniques, we screened and cloned 5 key enzyme gene atoB,hbd,crt,adhE2 and ter with relatively high activity in butanol synthesis pathway. Firstly, we constructed the pUC18-atoB-hbd-crt-ter plasmid by recombinant PCR, and then coloned tandem gene atoB-hbd-crt-ter and gene adhE2 into the plasmid pET21b and eventually constructed the expression vector of pET-21b-atoB-hbd-crt-ter-adhE2 containing butanol synthesis pathway, and transferred it into E.coli BL21 (DE3) for the preparation of engineering strains. We performed IPTG induced expression detection and gas chromatograph (GC) analysis of samples. The results show that key enzyme proteins have been synthesized and produced a certain amount of butanol.

Key words: escherichia coli (E.coli), biofuels, butanol, fermentation