Resveratrol Production from Hydrothermally Pretreated Eucalyptus Wood Using Recombinant Industrial Saccharomyces cerevisiae Strains
Resveratrol Production from Hydrothermally Pretreated Eucalyptus Wood Using Recombinant Industrial Saccharomyces cerevisiae Strains
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DOI：
10.1021/acssynbio.1c00120
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发表时间：
2021-07-24
影响因子：
4.7
通讯作者：
Domingues, Lucilia
中科院分区：
文献类型：
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作者：
Costa, Carlos E.;Moller-Hansen, Iben;Domingues, Lucilia
白藜芦醇是一种酚类化合物，具有很强的抗氧化活性，在保健品、食品和化妆品中具有广泛的应用前景。白藜芦醇通常从植物中提取或化学合成，过程复杂且不可持续，但白藜芦醇的微生物生物合成可以克服这些缺点。在这项工作中，白藜芦醇生产的微生物生物合成木质纤维素材料进行了评估。三个强大的工业酿酒酵母菌株已知的耐热性和/或耐抑制性化合物被确定为合适的主机从头白藜芦醇生产葡萄糖和乙醇。通过CRISPR/Cas9系统，所有工业菌株和实验室菌株都通过苯丙氨酸中间体成功地改造了白藜芦醇生物合成途径。所有菌株在30 ℃和39 ℃下进一步筛选以评估耐热性，这是同时发酵和发酵工艺的关键特征。乙醇红RBP在39 ℃下表现出最好的性能，与其他菌株相比，白藜芦醇产量超过2.6倍。然后使用该菌株来评估从葡萄糖和乙醇产生白藜芦醇。在含有2%葡萄糖和5%乙醇（w/v）的培养基中，白藜芦醇的最大滴度为187.07 +/- 19.88 mg/L。最后，乙醇红RBP在39摄氏度下，在同时澄清和发酵过程中，从来自水热预处理的蓝桉木材的2.95%的纤维素产生151.65 +/- 3.84 mg/L的白藜芦醇。据我们所知，这是木质纤维素白藜芦醇生产的第一份报告，为在工业背景下实施综合木质纤维素-白藜芦醇工艺奠定了基础。
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 degrees C and 39 degrees C to evaluate thermotolerance, which is a key feature for Simultaneous Saccharification and Fermentation processes. Ethanol Red RBP showed the best performance at 39 degrees 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 degrees 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.