WEEK 04

BIOPRODUCTION: LYCOPENE




Using tools such as Biocyc (https://www.biocyc.org) and KEGG (http://www.kegg.jp), review enzymes that can be added to (or knocked out of) your lycopene-producing E. coli to increase the amount of lycopene they produce. Compare the expected and experimental differences in production efficiency between bacteria that carry the pAC-LYC and pAC-LYCipi plasmids.



ENZYMES THAT CAN BE ADDED TO INCREASE: crtI - phytoene desaturase, crtQ - zeta-carotene desaturase, crtH - prolycopene isomerase

Using Biocyc and/or KEGG, can you identify the enzyme to convert the carrot-orange pigment beta-carotene to the corn-yellow pigment zeaxanthin?

ENZYMES CrtT and LUT5 CONVERT BETA CAROTENE TO BETA CRYPTOXANTHIN TO ZEAXANTHIN





1. SET UP OVERNIGHT CULTURES WITH 12 CONDITIONS, DUPLICATED.



2. CIRCULAR ROLLING DRUMS WITH 30 AND 37 DEGREES CELCIUS


3. OD 600 CELL COUNT



4. CONCENTRATE PELLET



5. ACETONE IN POLYPROPYLENE TUBES TO PREVENT REACTION


6. RESUSPEND PELLET IN ACETONE





7. EXTRACT PIGMENT



RESULTS




8. MEASURE ABSORPTION SPECTRUM