Centre of Biological Engineering, University of Minho, Campus Gualtar, 4710-057 Braga, Portugal.
The Novo Nordisk Foundation Center for Biosustainability, Technical University of Denmark, DK-2800 Kgs. Lyngby, Denmark.
ACS Synth Biol. 2021 Aug 20;10(8):1895-1903. doi: 10.1021/acssynbio.1c00120. Epub 2021 Jul 24.
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 strains known for their thermotolerance and/or resistance to inhibitory compounds were identified as suitable hosts for 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 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 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.
白藜芦醇是一种具有强抗氧化活性的酚类化合物，在健康、食品和化妆品等领域有广泛的应用前景。它通常从植物中提取，或者通过化学合成，这两种方法都很复杂，而且不可持续，但微生物生物合成白藜芦醇可以克服这些缺点。在这项工作中，评估了从木质纤维素材料通过微生物生物合成生产白藜芦醇。选择了三种具有热耐受性和/或对抑制性化合物抗性的工业菌株作为从葡萄糖和乙醇生产白藜芦醇的合适宿主。通过 CRISPR/Cas9 系统，成功地对所有工业菌株和一个实验室菌株进行了工程改造，使其具有白藜芦醇生物合成途径和苯丙氨酸中间产物。所有菌株都在 30°C 和 39°C 下进行了进一步筛选，以评估其热耐受性，这是同步糖化发酵过程的一个关键特征。Ethanol Red RBP 在 39°C 时表现最佳，与其他菌株相比，白藜芦醇产量提高了 2.6 倍以上。然后，该菌株被用于评估从葡萄糖和乙醇生产白藜芦醇。在含有 2%葡萄糖和 5%乙醇（w/v）的培养基中，最大白藜芦醇浓度达到 187.07±19.88mg/L。最后，在 39°C 下，Ethanol Red RBP 从经过水热预处理木材的纤维素中（含量为 2.95%）生产了 151.65±3.84mg/L 的白藜芦醇，在同步糖化发酵过程中。据我们所知，这是木质纤维素生产白藜芦醇的首次报道，为在工业环境中实施集成木质纤维素到白藜芦醇的过程奠定了基础。
