Engineering the oleaginous yeast Yarrowia lipolytica for high-level resveratrol production

Abstract

Resveratrol is a plant secondary metabolite with multiple health-beneficial properties. Microbial production of resveratrol in model microorganisms requires extensive engineering to reach commercially viable levels. Here, we explored the potential of the non-conventional yeast Yarrowia lipolytica to produce resveratrol and several other shikimate pathway-derived metabolites ( p -coumaric acid, cis , cis -muconic acid, and salicylic acid). The Y. lipolytica strain expressing a heterologous pathway produced 52.1 ± 1.2 mg/L resveratrol in a small-scale cultivation. The titer increased to 409.0 ± 1.2 mg/L when the strain was further engineered with feedback-insensitive alleles of the key genes in the shikimate pathway and with five additional copies of the heterologous biosynthetic genes. In controlled fed-batch bioreactor, the strain produced 12.4 ± 0.3 g/L resveratrol, the highest reported titer to date for de novo resveratrol production, with a yield on glucose of 54.4 ± 1.6 mg/g and a productivity of 0.14 ± 0.01 g/L/h. The study showed that Y. lipolytica is an attractive host organism for the production of resveratrol and possibly other shikimate-pathway derived metabolites.

Resveratrol Production in Yeast Hosts: Current Status and Perspectives

“…Conversely, strains harboring STS from Morusalba and P. cuspidatum accumulated resveratrol up to 39 mg/L using 70 mg/L p-coumaric acid [86]. Codon optimization for resveratrol pathway genes has also been observed in several studies in both S. cerevisiae and Y. lipolytica [88,91,93,95,96]. The highest resveratrol titer, which was produced by the expression of partial pathway genes (4CL and STS) in yeasts, was obtained from industrial Brazilian sugar-cane-fermenting yeast, and the titer was 391 mg/L of resveratrol [81].…”

“…Despite these efforts and extensive strain engineering, heterologous stilbene synthase activity and the overall resveratrol production are relatively still low. Except for one study that produced 12 g/L of resveratrol from Y. lipolytica [96], almost all engineered yeast strains do not produce more than 1 g/L, which does not meet the industrial needs and is the main drawback for their use on an industrial scale. Applying all of these elements together in a balanced way as well as testing other strategies that have been applied in E. coli may allow scientists to obtain the desired well-designed resveratrol-producing strain.…”

“…In addition to multiple integrations of resveratrol pathway genes (FjTAL, At4CL1, and VvVST1), feedback-insensitive YlARO4 K221L and YlARO7 G139S were employed to enhance resveratrol production. Further improvement was achieved by optimizing the growth medium and fed-batch fermentation [96].…”

Effect of Light and Dark on the Phenolic Compound Accumulation in Tartary Buckwheat Hairy Roots Overexpressing ZmLC

“…Many studies have focused on the enhancement of flavonoid production in bacteria [ 27 , 28 ], yeast [ 29 ], and plants [ 21 ]. Hairy roots from various plant species have been developed for the production of phenolic compounds because of their ability to produce large amounts of metabolites and their high growth rates [ 30 ].…”

Resveratrol Biosynthesis in Hairy Root Cultures of Tan and Purple Seed Coat Peanuts

“…Much effort has been made to increase the resveratrol yield in bacteria [26] that in the disccusion ed resveratrol contents compared with wild-type roots since rol gene co, yeast [27], algae [28], and plants [29]. Among the potential plant materials, hairy root cultures could be a very good source for the production of secondary metabolites, as these transgenic roots can produce the same specific metabolites as their mother plants, but in excessive amounts.…”