Denovo production of resveratrol by engineered Saccharomyces cerevisiae W303-1a using pretreated Gracilaria corticata extracts.

IF 4.6 Q2 MATERIALS SCIENCE, BIOMATERIALS ACS Applied Bio Materials Pub Date : 2024-02-01 Epub Date: 2023-11-21 DOI:10.1007/s10529-023-03441-4
Nishanthika Thenmozhi Kulasekaran, Mary Leema Thilakam, Dharani Gopal, Jung-Kul Lee, Jeya Marimuthu
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Abstract

Objective: Assembly and construction of resveratrol production pathway in Saccharomyces cerevisiae for denovo production of resveratrol using seaweed extract as fermentation medium.

Results: 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-1 respectively. CRISPR mediated integration at the δ locus resulted in 17.13 ± 1.1 mg resveratrol l-1. 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-1 when grown in seaweed extract medium.

Conclusions: The strain W303-1a/IntδC-res1 utilized Gracillaria hydrolysate and produced 13.6 ± 0.54 mg resveratrol l-1 and further investigations are being carried out focusing on pathway engineering and optimization of process parameters to enhance resveratrol yield.

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利用预处理的藤草提取物,利用工程酿酒酵母W303-1a生产白藜芦醇。
目的:以海藻提取物为发酵培养基,组装和构建酿酒酵母生产白藜芦醇的途径。结果:将酪氨酸途径产生白藜芦醇的相关基因、琼氏黄杆菌(Flavobacterium johnsoniae)的酪氨酸氨裂解酶(FTAL)基因、拟南芥(Arabidopsis thaliana)的4-香豆酸:辅酶a连接酶基因(4CL1)和葡萄(Vitis vinifera)的二苯乙烯合成酶基因(VvSTS)分别导入低拷贝、高拷贝和整合载体,转化为酿酒酵母W303-1a。所得菌株W303-1a/pARS-res5、W303-1a/2µ-res1和W303-1a/IntUra-res9的白藜芦醇l-1含量分别为2.39±0.01、3.33±0.03和8.34±0.03 mg。CRISPR介导的δ位点整合产生17.13±1.1 mg白藜芦醇l-1。以葛尾草提取物为底物,进行了白藜芦醇转化菌生长试验。该菌株在海藻提取液培养基中产生的白藜芦醇l-1含量为13.6±0.54 mg。结论:菌株w304 -1a/IntδC-res1利用江蓠的水解产物产白藜芦醇1(13.6±0.54 mg),可通过优化工艺参数和途径工程提高白藜芦醇的产率。
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来源期刊
ACS Applied Bio Materials
ACS Applied Bio Materials Chemistry-Chemistry (all)
CiteScore
9.40
自引率
2.10%
发文量
464
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