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Using carotenoid genes of Erwinia herbicola,metabolic engineering was carried out for lycopene productionwith the pAC-LYCO4 plasmid, which was composed of achromosomal DNA fragment of E. herbicolacrtE, crtB, and crtI genes under the control of the tetracyclinepromoter and the ipi gene of Haematococcus pluvialis withthe trc promoter. Plasmid pAC-LYCm4 was constructed forefficient expression of the four exogenous genes using astrong RBS sequence and the same tetracycline promoter. Theoptimized expression construct of pAC-LYCm4 increasedlycopene production thre times as compared with pAC-LYCO4. pAC-LYCm5 containing ispAgenes was constructed. There was no significant diference inlycopene production and cell growth between pAC-LYCm4and pAC-LYCm5. FPP synthase encoded by ispA was notrate-limiting for lycopene production. Each gene of crtE,crtB, crtI, and ipi was overexpresed, using pBAD-crtE,pBAD-crtIB, and pBAD-ipiHP1, in addition to their expressionfrom pAC-LYCm4. However, there was no increase of lycopeneproduction with the additional overexpression of each exogenousgene. The four exogenous genes appeared to be not rate-limiting in cells harboring pAC-LYCm4. When pDdxs, pBAD24containing dxs, was introduced into cels harboring lycopenesynthetic plasmids, lycopene production of pAC-LYCO4,pAC-LYCm4, and pAC-LYCm5 was increased by 4.7-, 2.2-,and 2.2-fold, respectively. Lycopene production of pBAD-DXm4 containing crtE, crtB, crtI, ipi, and dxs was 5.2 mg/gdry cell weight with 0.2% arabinose, which was 8.7-fold higherthan that of the initial strain with pAC-LYCO4. Therefore, thepresent study showed that proper regulation of a metabolically

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