Week 12 Lab: Bioproduction of Beta-Carotene and Lycopene

Bioproduction of Beta-Carotene and Lycopene Lab

• Was unable to perform this Lab protocol at the William & Mary Node Wet Lab, as this protocol was not performed at the Lab as part of William & Mary’s HTGAA engagement this semester. However I did review the related ‘Bioproduction of Beta-Carotene and Lycopene Lab’ protocol. Answers to the questions found in the ‘Post Lab Questions’ section of the protocol can be found below:

  • Which genes when transferred into E. coli will induce the production of lycopene and beta-carotene, respectively?
    • The Erwinia herbicola crtE, crtI, crtB, and crtY genes induce respective lycopene and beta-carotene production
  • Why do the plasmids that are transferred into the E. coli need to contain an antibiotic resistance gene?
    • Antibiotic resistance genes help researchers identify which E. coli bacteria successfully took the plasmid. This is necessary because a lot of the time E. coli bacteria do not successfully take plasmids
  • Whast outcomes might we expect to see when we vary the media, presence of fructose, and temperature conditions of the overnight cultures?
    • We might expect different levels of lycopene and beta-carotene production (i.e., different levels of biosynthesis, difficult absorption, and/or potentially different shades of produced pigments by varying overnight culture media, presence of fructose, and temperature conditions
  • Generally describe what “OD600” measures and how it can be interpreted in this experiment
    • OD600 measuress cell concentration and peak absorption for respective samples post-cellular culture incubation. It can be interpreted to determine whether or not we actually produced the desired forms of lycopene and beta-carotene with the appropriate pigmentation because we can compare our experimental absorption results form our cultures with previously established lycopene and beta-carotene absorption results in the literature
  • What are other experimental setups where we may be able to use acetone to separate cellular matter from a compound we intend to measure?
    • It would appear that acetone would be useful for experimental setups like separating out certain proteins from a larger mixture
  • Why might we want to engineer E. coli to produce lycopene and beta-carotene pigments when Erwinia herbicola naturally produces them?
    • Likely because we want more control over production of said pigments than Erwinia herbicola naturally provides. Control in this case might extend to pigment shade, concentration, or production time

All supporting prompts for this section listed below