Protocols for the Production and Analysis of Isoprenoids in Bacteria and Yeast

Isoprenoids (a.k.a. terpenes/terpenoids) are the largest group of natural products. They fulfil a wide variety of both essential and non-essential roles in biology; many isoprenoids also have useful industrial applications. In recent years, there has been a significant focus on metabolic engineering of various isoprenoids in microbial cells. Here, we describe methods for lab-scale culturing, sampling and analytics of different isoprenoid classes using specific examples from the following classes: (1) highly volatile isoprenoids that sequester into culture headspaces, e.g. the hemiterpene isoprene; (2) volatile isoprenoids that can be collected in a non-toxic organic phase, e.g. the monoterpene limonene; and (3) non-volatile isoprenoids that accumulate in the cell, e.g. the carotenoid lycopene. Production methods are provided for isoprene, limonene, and lycopene as examples for each class. Specific analytical methods are provided for isoprene, limonene, terpinolene, caryophyllene, amorphadiene, linalool, nerolidol, pinene and lycopene. We focus on yeast and Escherichia coli as production organisms. The protocols can be modified for other organisms and products as appropriate.