Biosynthesis of ursolic acid and oleanolic acid in Saccharomyces cerevisiae

Abstract

Ursolic acid and oleanolic acid are pentacyclic triterpenoid compounds with a variety of biological activities. A mixture of ursolic acid and oleanolic acid has higher antitumor activity than the individual acids. We have developed a simultaneous biosynthesis pathway for different proportions of ursolic acid and oleanolic acid in Saccharomyces cerevisiae. The ScLCZ08 strain produced 175.15 mg/L of the ursolic acid precursor α‐amyrin, the highest amount reported. Ursolic and oleanolic acid titers and proportions were optimized using Medicago truncatula amyrin C‐28 oxidase and Arabidopsis thaliana cytochrome P450 reductase. Using glucose and ethanol fed‐batch fermentation strategies, the final ursolic acid and oleanolic acid titers were 123.27 and 155.58 mg/L, respectively, demonstrating 4.77‐fold and 4.95‐fold higher production than the parent strain. The ScLCZ11 strain displayed the highest ursolic acid production obtained via microbial fermentation in fed‐batch culture to date. © 2018 American Institute of Chemical Engineers AIChE J, 64: 3794–3802, 2018

Identifying and engineering the ideal microbial terpenoid production host

“…Dammarenediol II production has also been described in E. coli , requiring introduction of heterologous 2,3-oxidosqualene biosynthesis as well truncation of all N-terminal transmembrane domains of involved enzymes (Li et al 2016) and P. pastoris (Zhao et al 2016) but, in comparison with S. cerevisiae , at relatively low titers and specific yields of 8.63 mg L −1 and 1.04 mg g −1 DCW, respectively. α- and β-amyrin as well as their CYP450-derived products ursolic and oleanolic acid have been obtained in the low 3-digit mg L −1 range in S. cerevisiae (Lu et al 2018). Recently, Zhao et al (2018) enhanced oleanolic acid levels to 607 mg L −1 in S. cerevisiae .…”

Biotransformation of Oleanane and Ursane Triterpenic Acids

“…Because no enzymes were found to synthesize exclusively α- Genes encoding β-amyrin synthase (βAS) that catalyze the formation of β-amyrin (6)-a precursor of oleanane pentacyclic triterpenoids-from 2,3-oxidosqualene (5) were isolated from the genomes of Glycyrrhiza glabra, Panax ginseng, Catharanthus roseus, Lotus japonicus, Artemisia annua, Chenopodium quinoa, and others [65][66][67][68][69]. Because no enzymes were found to synthesize exclusively α-amyrin (7), a precursor of ursane pentacyclic triterpenoids, this reaction involves mixed amyrin synthases (mix-AS) that catalyze the formation of both αand β-amyrin from Eriobotrya japonica and C. roseus [64,70]. Medicago truncatula is most often used to search for genes encoding CYP450 enzymes that catalyze subsequent conversion of αand β-amyrin [64][65][66][67]69,71].…”

Preparative separation of structural isomeric pentacyclic triterpene oleanolic acid and ursolic acid from natural products by pH-zone-refining countercurrent chromatography

“…According to reports in literature, the mixture of OA and UA had a higher pharmacological activity compared with the activity observed with either OA or UA alone. 16,[35][36][37] The interaction of OA and UA may have taken place chemically and biologically to produce an ideal effect. 38 Therefore, pharmacological research on the mixture of OA and UA was conducted by numerous researchers.…”