Increased beta-carotene production in recombinant Escherichia coli harboring an engineered isoprenoid precursor pathway with mevalonate addition.
When pT-LYCm4 containing lycopene synthetic genes was co-transformed with pSUcrtY or pSHcrtY containing crtY gene of Pantoea ananatis (P. ananatis) or Pantoea agglomerans (P. agglomerans), beta-carotene productions of 36 and 35 mg/L were obtained, respectively. No lycopene was detected in the beta-carotene production culture. pT-HB, constructed by addition of P. ananatis crtY gene into pT-LYCm4, was used for co-transformation with pSdxs and pSSN12Didi, which increased isopentenyl diphosphate and dimethylallyl diphosphate synthesis. beta-Carotene production significantly increased 1.5-fold (51 mg/L) with the amplification of the dxs gene through pSdxs and 4-fold (135 mg/L) with the mevalonate bottom pathway of pSSN12Didi in the presence of 3.3 mM mevalonate. The pT-DHB, constructed by integrating the dxs gene into pT-HB, was used for cotransformation of Escherichia coli (E. coli) harboring pSSN12Didi, resulting in beta-carotene production of 141 mg/L. Recombinant E. coli harboring pT-DHB and pSSN12Didi was used to maximize beta-carotene production by adjusting the available amounts of glycerol, a carbon source, and mevalonate, the precursor of the mevalonate bottom pathway. When recombinant E. coli was given 16.5 mM mevalonate and 2.5% (w/v) glycerol, beta-carotene production of 503 mg/L in concentration and 49.3 mg/g DCW in content was obtained at 144 h, which was the highest level of carotenoid production in E. coli ever reported in the literature.
Yoon
Sang-Hwal
SH
Division of Applied Life Science (BK21), EB-NCRC and PMBBRC, and Division of Forest Science, Gyeongsang National University, Jinju 660-701, Korea.
Park
Hye-Min
HM
Kim
Ju-Eun
JE
Lee
Sook-Hee
SH
Choi
Myung-Suk
MS
Kim
Jae-Yean
JY
Oh
Deok-Kun
DK
Keasling
Jay D
JD
Kim
Seon-Won
SW
D001426:Bacterial Proteins / Q000235:genetics / Q000378:metabolism; D004926:Escherichia coli / Q000187:drug effects / Q000235:genetics* / Q000378:metabolism; D015964:Gene Expression Regulation, Bacterial / Q000187:drug effects; D005818:Genetic Engineering / Q000379:methods; D005990:Glycerol / Q000494:pharmacology; D045782:Hemiterpenes / Q000378:metabolism; D019753:Intramolecular Lyases / Q000235:genetics* / Q000378:metabolism; D008798:Mevalonic Acid / Q000494:pharmacology*; D008954:Models, Biological; D009943:Organophosphorus Compounds / Q000378:metabolism; D020636:Pantoea / Q000235:genetics; D013729:Terpenes / Q000378:metabolism*; D019207:beta Carotene / Q000378:metabolism*
D001426:Bacterial Proteins; D045782:Hemiterpenes; D009943:Organophosphorus Compounds; D013729:Terpenes; D019207:beta Carotene; C004809:isopentenyl pyrophosphate; D019753:Intramolecular Lyases; C070925:lycopene cyclase-isomerase; D005990:Glycerol; D008798:Mevalonic Acid