Production of miltiradiene by metabolically engineered Saccharomyces cerevisiae

	Production of miltiradiene by metabolically engineered Saccharomyces cerevisiae
	Dai, Zhubo 2; Liu, Yi 2,3; Huang, Luqi 1; Zhang, Xueli 2
刊名	BIOTECHNOLOGY AND BIOENGINEERING
	2012-11-01
卷号	109 期号:11 页码:2845-2853
关键词	tanshinones miltiradiene terpenoids isoprenoid precursors fermentation Saccharomyces cerevisiae
英文摘要	Metabolic engineering of microorganisms is an alternative and attractive route for production of valuable terpenoids that are usually extracted from plant sources. Tanshinones are the bioactive components of Salvia miltiorrhizha Bunge, which is a well-known traditional Chinese medicine widely used for treatment of many cardiovascular diseases. As a step toward microbial production of tanshinones, copalyl diphosphate (CPP) synthase, and normal CPP kaurene synthase-like genes, which convert the universal diterpenoid precursor geranylgeranyl diphosphate (GGPP) to miltiradiene (an important intermediate of the tanshinones synthetic pathway), was introduced into Saccharomyces cerevisiae, resulting in production of 4.2?mg/L miltiradiene. Improving supplies of isoprenoid precursors was then investigated for increasing miltiradiene production. Although over-expression of a truncated 3-hydroxyl-3-methylglutaryl-CoA reductase (tHMGR) and a mutated global regulatory factor (upc2.1) gene did improve supply of farnesyl diphosphate (FPP), production of miltiradiene was not increased while large amounts of squalene (78?mg/L) were accumulated. In contrast, miltiradiene production increased to 8.8?mg/L by improving supply of GGPP through over-expression of a fusion gene of FPP synthase (ERG20) and endogenous GGPP synthase (BTS1) together with a heterologous GGPP synthase from Sulfolobus acidocaldarius (SaGGPS). Auxotrophic markers in the episomal plasmids were then replaced by antibiotic markers, so that engineered yeast strains could use rich medium to obtain better cell growth while keeping plasmid stabilities. Over-expressing ERG20-BTS1 and SaGGPS genes increased miltiradiene production from 5.4 to 28.2?mg/L. Combinatorial over-expression of tHMGR-upc2.1 and ERG20-BTS1-SaGGPS genes had a synergetic effects on miltiradiene production, increasing titer to 61.8?mg/L. Finally, fed-batch fermentation was performed, and 488?mg/L miltiradiene was produced. The yeast strains engineered in this work provide a basis for creating an alternative way for production of tanshinones in place of extraction from plant sources. Biotechnol. Bioeng. 2012; 109: 28452853. (c) 2012 Wiley Periodicals, Inc.
WOS标题词	Science & Technology ; Life Sciences & Biomedicine
类目[WOS]	Biotechnology & Applied Microbiology
研究领域[WOS]	Biotechnology & Applied Microbiology
关键词[WOS]	YEAST ; PATHWAY ; OVERPRODUCTION ; PRECURSOR ; SYNTHASE ; OVEREXPRESSION ; BIOSYNTHESIS ; ACCUMULATION ; VECTORS ; CLONING
收录类别	SCI
语种	英语
WOS记录号	WOS:000309185600016
公开日期	2014-11-19
内容类型	期刊论文
源URL	[http://124.16.173.210/handle/311007/364]
专题	天津工业生物技术研究所_微生物代谢工程张学礼_期刊论文
作者单位	1.Chinese Acad Chinese Med Sci, Inst Chinese Mat Med, Beijing 100700, Peoples R China 2.Chinese Acad Sci, Key Lab Syst Microbial Biotechnol, Tianjin Inst Ind Biotechnol, Tianjin 300308, Peoples R China 3.Tianjin Univ Sci & Technol, Coll Biotechnol, Tianjin, Peoples R China
推荐引用方式 GB/T 7714	Dai, Zhubo,Liu, Yi,Huang, Luqi,et al. Production of miltiradiene by metabolically engineered Saccharomyces cerevisiae[J]. BIOTECHNOLOGY AND BIOENGINEERING,2012,109(11):2845-2853.
APA	Dai, Zhubo,Liu, Yi,Huang, Luqi,&Zhang, Xueli.(2012).Production of miltiradiene by metabolically engineered Saccharomyces cerevisiae.BIOTECHNOLOGY AND BIOENGINEERING,109(11),2845-2853.
MLA	Dai, Zhubo,et al."Production of miltiradiene by metabolically engineered Saccharomyces cerevisiae".BIOTECHNOLOGY AND BIOENGINEERING 109.11(2012):2845-2853.