Part:BBa_K1604020
araC-pBAD + beta-carotene
This device produces β carotene under the control of an arabinose promoter.
Usage and Biology
This device contains the four genes necessary for βcarotene biosynthesis.
FIGURE 1. Biochemical pathway of βcarotene. βcarotene biosynthesis from pharnesyl di phospahate a colorless molecule naturally produced in E . coli.
FIGURE 2. Production of β-carotene. NEB10β cells were transformed with BBa_K1604020 and grown up to an OD of 0.6 and induced with 5 mM of arabinose for 24 hours. BBa_K1604020 (β carotene) induced (A) and not induced (B); Also uninduced cells produced high amounts of βcarotene.
FIGURE 3. Extraction of β-carotene. NEB10β cells were transformed with BBa_K1604020 and grown in 100 mL of LB. The cells were induced with 5 mM of arabinose and for 24 hours. The cells were span down and the supernatant was discarded. The pellets were incubated with 2.5 mL of acetone for 10 min at 50C. Afterward they were centrifuged to recover the extracted pigments. Panel A. extraction in acetone of βcarotene. Panel B. TLC analysis of βcarotene reference and BBa_K1604020 induced and uninduced with arabinose.
FIGURE 4. UV VIS spectra of carotenoids. The pigment were extracted as described in Figure 3. The extracted samples were diluted 1:7 in acetone and the spectra were taken with UV-VIS Agilent. The spectra were acquired between 300 and 800 nm and blanked with acetone.Panel B UV VIS spectra: reference βcarotene (..); BBaK173201, aracpbad (..) and BBa_K1604020 (aracpbad- β carotene) with 5 mM arabinose.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21INCOMPATIBLE WITH RFC[21]Illegal BamHI site found at 1144
Illegal BamHI site found at 3185 - 23COMPATIBLE WITH RFC[23]
- 25INCOMPATIBLE WITH RFC[25]Illegal NgoMIV site found at 2721
Illegal NgoMIV site found at 2851
Illegal AgeI site found at 979
Illegal AgeI site found at 1936 - 1000INCOMPATIBLE WITH RFC[1000]Illegal SapI site found at 961