HTGAA Class 4a: Bioproduction

​The purpose of this week’s lab is to test the impact of culture conditions (i.e., culture temperature and growth media composition) on the production of lycopene and beta-carotene in E. coli. 

​​Lycopene is the red pigment that gives tomatoes their red color. This pigment is also made by microbes. In fact, transferring a 3-enzyme pathway to E. coli can convert farnesyl diphosphate (FPP) to lycopene. The computational tools and databases presented today can also be used to enhance lycopene production in E.coli or even produce different colors, such as the beta-carotene pigment that makes carrots orange.

​​For the laboratory portion of this assignment, you will characterize lycopene production in E.coli with thepAC-LYC plasmid. You will also characterize beta-carotene production in E.coli with thepAC-BETA plasmid. Note, the pAC-LYC plasmid contains three genes from Erwinia herbicola: CrtE, Crtl, and CrtB. The pAC-BETA plasmid produces beta-carotene through the addition of the Erwinia herbicola CrtY gene. All plasmids include the gene for chloramphenicol resistance.

​As an extension upon this lab, we will be working with the pioreactor to determine proper culture conditions for manufacturing colorants.