Obtaining 2,3-Dihydrobenzofuran and 3-Epilupeol from Ageratina pichinchensis (Kunth) R.King & Ho.Rob. Cell Cultures Grown in Shake Flasks under Photoperiod and Darkness, and Its Scale-Up to an Airlift Bioreactor for Enhanced Production

Abstract

Ageratina pichinchensis (Kunth) R.King & Ho.Rob. is a plant used in traditional Mexican medicine, and some biotechnological studies have shown that its calluses and cell suspension cultures can produce important anti-inflammatory compounds. In this study, we established a cell culture of A. pichinchensis in a 2 L airlift bioreactor and evaluated the production of the anti-inflammatory compounds 2,3-dihydrobenzofuran (1) and 3-epilupeol (2). The maximum biomass production (11.90 ± 2.48 g/L) was reached at 11 days of culture and cell viability was between 80% and 90%. Among kinetic parameters, the specific growth rate (µ) was 0.2216 days−1 and doubling time (td) was 3.13 days. Gas chromatography coupled with mass spectrometry (GC-MS) analysis of extracts showed the maximum production of compound 1 (903.02 ± 41.06 µg/g extract) and compound 2 (561.63 ± 10.63 µg/g extract) at 7 and 14 days, respectively. This study stands out for the significant production of 2,3-dihydrobenzofuran and 3-epilupeol and by the significant reduction in production time compared to callus and cell suspension cultures, previously reported. To date, these compounds have not been found in the wild plant, i.e., its production has only been reported in cell cultures of A. pichinchensis. Therefore, plant cell cultured in an airlift reactor can be an alternative for the improved production of these anti-inflammatory compounds.

Cytotoxic Activity of Wild Plant and Callus Extracts of Ageratina pichinchensis and 2,3-Dihydrobenzofuran Isolated from a Callus Culture

“…On the other hand, the compounds stigmasterol (5), stigmasterol glucoside (11), and β-sitosterol (6) are considered some of the predominant compounds in plants [50,51]. Their pharmacological studies have shown their antimicrobial, anti-inflammatory, antiosteoarthritic, anti-tumor, cytotoxic, anti-hypercholesterolemia, and antioxidant activities [52][53][54][55].…”

“…Moreover, compound (7) exhibits chirality, which complicates the development of synthesis routes to obtain it. On the other hand, our research group has reported the production of the compound through suspension cell cultures in flasks as well as in airlift reactors, significantly improving its production and possible scaling [10,11], which provides a viable and sustainable alternative to produce the compound.…”

“…The GE-4 fraction was made up of α-amyrin (8), 3-epilupeol (9), and campesterol (10). Compound (11) was identified in the GE-5 fraction by direct comparison with an authentic sample available in the laboratory.…”

“…For instance, the production of the compounds 3-epilupeol and a benzofuran was found in callus and cell suspension cultures, which are not produced by wild plants. In fact, the production of these anti-inflammatory compounds could be improved using an airlift reactor, and their production was improved by 1.82-to 1.35-fold, respectively [9][10][11].…”