Microbial Production, Extraction, and Quantitative Analysis of Isoprenoids

Satta, Alessandro; Lu, Zeyu; Plan, Manuel R.; Esquirol, Lygie; Ebert, Birgitta E.

doi:10.1007/978-1-0716-2185-1_20

Cited by 1 publication

(1 citation statement)

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“…Triterpenoids were quantified by HPLC-CAD following a published protocol. 43 Specifically, samples of 800 μL of culture broth were transferred into 2 mL sample tubes to analyze intracellular triterpenoid concentrations and were stored at -20 °C until further processing or immediately analyzed. Cells were disrupted by adding 250 μL of glass beads (smooth 500 μm, Carl Roth, Karlsruhe, Germany), 80 μL 1 M HCl, and 800 μL chloroform: methanol (ratio 80:20, v/v) and vigorously agitating in the Mini-Beadbeater 16 (BioSpec Products, Bartlesville, USA) for 4 min.…”

Section: Methodsmentioning

confidence: 99%

Triterpenoid production with a minimally engineered Saccharomyces cerevisiae chassis

Guo

Jacobsen

Walter

et al. 2022

Preprint

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Triterpenoids, one of the most diverse classes of natural products, have been used for centuries as active ingredients in essential oils and Chinese medicines and are of interest for many industrial applications ranging from low-calorie sweeteners to cosmetic ingredients and vaccine adjuvants. However, not only can the extraction from plant material be cumbersome due to low concentrations of the specific triterpenoid, but concerns are also increasing regarding the sustainability of wild plant harvest while meeting market demands. The alternative is to produce triterpenoids with engineered microbes. Here, we present a generally applicable strategy for triterpenoid production in the yeast Saccharomyces cerevisiae based on a modified oxidosqualene cyclase Erg7. The modification reduces the flux into the sterol pathway while increasing the precursor supply for triterpenoid production. The minimally engineered strain was exploited for the exemplary production of the lupane triterpenoids betulin, betulin aldehyde, and betulinic acid at a total titer above 6 g/L, the highest reported so far. To further highlight the chassis concept, squalene, oleanane- and dammarane-type triterpenoids were synthesized to titers at a similar gram scale. We propose the developed baker's yeast as a host for the thousands of triterpenoid synthesis pathways from plants, reducing the pressure on the natural resources.

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Section: Methodsmentioning

confidence: 99%