Difference between revisions of "Part:BBa K1604020"

Revision as of 17:53, 16 September 2015

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. html><a href="#fn:1" id="fnref:1" title="see footnote" class="footnote">[1]</a></html>

FIGURE 1. Biochemical pathway of β-carotene. 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. Cells transformed with BBa_K731201(araC-pBAD) were used as negative control for β-carotene production. BBa_K731201(araC-pBAD) (A), BBa_K1604020 (β-carotene producer) without arabinose 5mM(B), and not induced (C). 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 50 °C.[2] Afterward they were centrifuged to recover the extracted pigments. Panel A. extraction in acetone of β-carotene. Panel B. TLC analysis of β extract from BBa_K1604020 (β-carotene producer) without arabinose 5mM (A), and induced (B) β-carotene reference (C).

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 (green); BBa_K173201, control cells (violet): and BBa_K1604020 (aracpbad- β-carotene) with 5 mM arabinose (red) and withouy induction(blue).</p> Check out our Wiki UNITN-Trento iGEM 2015!