Marine Biotechnology Division, National Institute of Ocean Technology, Chennai, 600100, India.
Department of Chemical Engineering, Konkuk University, Seoul, 143 701, Korea.
Biotechnol Lett. 2024 Feb;46(1):19-28. doi: 10.1007/s10529-023-03441-4. Epub 2023 Nov 21.
Assembly and construction of resveratrol production pathway in Saccharomyces cerevisiae for denovo production of resveratrol using seaweed extract as fermentation medium.
Genes involved in the production of resveratrol from tyrosine pathway, tyrosine ammonia lyase (FTAL) gene from Flavobacterium johnsoniae (FjTAL), the 4-coumarate:CoA ligase gene from Arabidopsis thaliana (4CL1) and the stilbene synthase gene from Vitis vinifera (VvSTS) were introduced into low copy, high copy and integrative vector and transformed into S. cerevisiae W303-1a. The resulting strains W303-1a/pARS-res5, W303-1a/2µ-res1 and W303-1a/IntUra-res9 produced a level of 2.39 ± 0.01, 3.33 ± 0.03 and 8.34 ± 0.03 mg resveratrol l respectively. CRISPR mediated integration at the δ locus resulted in 17.13 ± 1.1 mg resveratrol l. Gracilaria corticata extract was tested as a substrate for the growth of transformant to produce resveratrol. The strain produced a comparable level, 13.6 ± 0.54 mg resveratrol l when grown in seaweed extract medium.
The strain W303-1a/IntδC-res1 utilized Gracillaria hydrolysate and produced 13.6 ± 0.54 mg resveratrol l and further investigations are being carried out focusing on pathway engineering and optimization of process parameters to enhance resveratrol yield.
在酿酒酵母中组装和构建白藜芦醇生产途径，以利用海藻提取物作为发酵培养基从头生产白藜芦醇。
从黄杆菌（FjTAL）中引入酪氨酸途径生产白藜芦醇的相关基因、拟南芥（4CL1）的 4-香豆酸：CoA 连接酶基因和葡萄（VvSTS）的芪合酶基因到低拷贝、高拷贝和整合载体中，并转化到酿酒酵母 W303-1a 中。得到的菌株 W303-1a/pARS-res5、W303-1a/2µ-res1 和 W303-1a/IntUra-res9 分别产生 2.39±0.01、3.33±0.03 和 8.34±0.03 mg l 的白藜芦醇。在 δ 位点进行 CRISPR 介导的整合导致 17.13±1.1 mg l 的白藜芦醇。测试了石花菜提取物作为转化体生长的底物来生产白藜芦醇。当在海藻提取物培养基中生长时，该菌株产生了相当水平的 13.6±0.54 mg l 白藜芦醇。
菌株 W303-1a/IntδC-res1 利用石花菜水解物并产生 13.6±0.54 mg l 白藜芦醇，正在进行进一步的研究，重点是途径工程和优化工艺参数以提高白藜芦醇产量。
