Resveratrol Production from Hydrothermally Pretreated Eucalyptus Wood Using Recombinant Industrial Saccharomyces cerevisiae Strains

Abstract

Resveratrol is a phenolic compound with strong antioxidant activity, being promising for several applications in health, food, and cosmetics. It is generally extracted from plants or chemically synthesized, in both complex and not sustainable processes, but microbial biosynthesis of resveratrol can counter these drawbacks. In this work, resveratrol production by microbial biosynthesis from lignocellulosic materials was assessed. Three robust industrial Saccharomyces cerevisiae strains known for their thermotolerance and/or resistance to inhibitory compounds were identified as suitable hosts for de novo resveratrol production from glucose and ethanol. Through the CRISPR/Cas9 system, all industrial strains, and a laboratory one, were successfully engineered with the resveratrol biosynthetic pathway via the phenylalanine intermediate. All strains were further screened at 30 °C and 39 °C to evaluate thermotolerance, which is a key feature for Simultaneous Saccharification and Fermentation processes. Ethanol Red RBP showed the best performance at 39 °C, with more than 2.6-fold of resveratrol production in comparison with the other strains. This strain was then used to assess resveratrol production from glucose and ethanol. A maximum resveratrol titer of 187.07 ± 19.88 mg/L was attained from a medium with 2% glucose and 5% ethanol (w/v). Lastly, Ethanol Red RBP produced 151.65 ± 3.84 mg/L resveratrol from 2.95% of cellulose from hydrothermally pretreated Eucalyptus globulus wood, at 39 °C, in a Simultaneous Saccharification and Fermentation process. To the best of our knowledge, this is the first report of lignocellulosic resveratrol production, establishing grounds for the implementation of an integrated lignocellulose-to-resveratrol process in an industrial context.

Engineered biosynthesis of plant polyketides by type III polyketide synthases in microorganisms

“…After enhancing P450 activity, overexpressing pathway genes, and increasing the precursor supply, a final production of 800 mg/L was achieved in fed-batch fermentation from glucose. In 2021, Costa et al (2021) used a mixed medium with 2% glucose and 5% ethanol to produce resveratrol and achieved a maximum 187.07 ± 19.88 mg/L production. Multiple similar strategies used in naringenin production were also applied in resveratrol production in microbial hosts to obtain higher production (Wu et al, 2017b;Hong et al, 2020;Sáez-Sáez et al, 2020;Yuan et al, 2020;Li et al, 2022).…”

Valorisation of wine wastes by de novo biosynthesis of resveratrol using a recombinant xylose-consuming industrial Saccharomyces cerevisiae strain

“…Our observation is in accordance with previous studies on de novo resveratrol production from glucose. 7,8 On the other hand, while simultaneously fermenting glucose and xylose, the yeast achieved approx. 48% of the overall pro- duction in the first 12 h, increasing the resveratrol yield of the sugar phase.…”

“…1 Even though therapeutical effects of resveratrol in humans are still unclear regarding its mode of action and molecular target, several beneficial properties are attributed to resveratrol, such as treatment/prevention of cardiovascular diseases, 2 and antiinflammatory 3 and anti-ageing properties. 4,5 Microbial production of resveratrol from carbon sources can be attained through tyrosine 6 or phenylalanine [7][8][9] via the shikimate pathway. In the latter, phenylalanine is converted into p-coumaric acid through the intermediary cinnamic acid.…”

“…Resveratrol production has been previously reported in engineered Saccharomyces cerevisiae from different carbon sources such as glucose, lactose or ethanol. [6][7][8][9] The current environmental and economic challenges have pushed the need for sustainable alternatives to produce energy and value-added products from renewable resources. 11,12 In this sense, using agro-industrial wastes has been gaining attention as possible feedstocks for biotechnological processes.…”

Synthetic Biology-Driven Microbial Production of Resveratrol: Advances and Perspectives

“…It is noteworthy that the use of microorganisms has made significant contributions to the biosynthesis of pharmaceutical and industrial compounds in recent decades, because of its low production cost, high efficiency, high product purity and simple genetic operation process, and it is considered to have a promising potential for the production of natural products ( Cravens et al, 2019 ). De novo resveratrol biosynthesis via metabolic engineering and synthetic biology in microorganisms provides a feasible way to produce resveratrol, and this has attracted worldwide interest ( Liu et al, 2019 ; He et al, 2020 ; Liu Z et al, 2020 ; Yuan et al, 2020 ; Costa et al, 2021 ). This review discusses the current status and progress of resveratrol production in recent years, as well as optimization strategies for related hosts, pathways and enzymes for resveratrol production.…”