Week 12 Lab: Bioproduction of Beta-Carotene and Lycopene

Questions etc. are here: https://2026a.htgaa.org/2026a/course-pages/weeks/week-12/lab/index.html

Post Lab Questions | Mandatory for All Students

  1. Which genes when transferred into E. coli will induce the production of lycopene and beta-carotene, respectively?
  2. Why do the plasmids that are transferred into the E. coli need to contain an antibiotic resistance gene?
  3. What outcomes might we expect to see when we vary the media, presence of fructose, and temperature conditions of the overnight cultures?
  4. Generally describe what “OD600” measures and how it can be interpreted in this experiment.

OD600 is optical density at 600 nanometer wavelngth … this helps measure how cloudy the liquid is, to determine cell growth.

  1. What are other experimental setups where we may be able to use acetone to separate cellular matter from a compound we intend to measure?

We could also use acetone to separate out proteins …

  1. Why might we want to engineer E. coli to produce lycopene and beta-carotene pigments when Erwinia herbicola naturally produces them?

Post Lab Questions | For Committed Listeners Only

Let’s get in touch with our metabolic pathway!

  1. What are the enzymes of the carotene pathway?
  2. Within this pathway, which is the rate determining step (the step that takes the longest)? Which enzyme is responsible for this step?

Notes for design of a DNA construct for bioproduction

  1. The first thing to do is to decide what organism you are going to use for this (E. coli or S. cerevisiae) for production. Which would you choose and why (emphases on production differences)?
  2. Now choose one of the enzymes and lets outline the parts of the construct for expression.
  3. For E. coli lets create a expression vector that works as a plasmid you choose E. coli let’s create a expression vector that works as a plasmids