Dual regulation of lipid droplet-triacylglycerol metabolism and ERG9 expression for improved β-carotene production in Saccharomyces cerevisiae.
Dual regulation of lipid droplet-triacylglycerol metabolism and ERG9 expression for improved β-carotene production in Saccharomyces cerevisiae.
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DOI：
10.1186/s12934-021-01723-y
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发表时间：
2022-01-04
影响因子：
6.4
通讯作者：
Yan GL
中科院分区：
文献类型：
--
作者：
Bu X;Lin JY;Duan CQ;Koffas MAG;Yan GL
非胡萝卜素工程菌高效高产类胡萝卜素面临着储存空间的限制、产物的细胞毒性和前体竞争等问题。为了提高酿酒酵母细胞中β-胡萝卜素的积累，本研究采用外源油酸（OA）结合代谢工程的方法，在提高细胞贮藏能力的同时，强化了类胡萝卜素途径的代谢通量。脂滴的直接分离、定量分析和基因阻断试验表明，脂滴是β-胡萝卜素在S.酿酒。但是，由于β-胡萝卜素与LDs-三酰甘油生物合成之间存在前体竞争，通过改造LDs相关基因扩大贮藏空间对β-胡萝卜素积累的促进作用不大。添加2 mM OA显著促进了LD-三酰甘油代谢，导致β-胡萝卜素含量增加36.4%。利用转录组分析方法挖掘OA抑制性启动子，并以IZH 1启动子替代ERG 9启动子，动态下调ERG 9的表达，使代谢流转向β-胡萝卜素途径，在不影响细胞生长的情况下，使β-胡萝卜素含量增加31.7%。通过诱导一条额外的组成型β-胡萝卜素合成途径将前体法尼醇进一步转化为β-胡萝卜素，最终菌株的DCW产量为11.4mg/g，β-胡萝卜素产量为142 mg/L，分别比出发菌株提高了107.3%和49.5%。该策略可应用于在S.酿酒。在线版本包含补充材料，可通过10.1186/s12934-021-01723-y获得。
The limitation of storage space, product cytotoxicity and the competition for precursor are the major challenges for efficiently overproducing carotenoid in engineered non-carotenogenic microorganisms. In this work, to improve β-carotene accumulation in Saccharomyces cerevisiae, a strategy that simultaneous increases cell storage capability and strengthens metabolic flux to carotenoid pathway was developed using exogenous oleic acid (OA) combined with metabolic engineering approaches. The direct separation of lipid droplets (LDs), quantitative analysis and genes disruption trial indicated that LDs are major storage locations of β-carotene in S. cerevisiae. However, due to the competition for precursor between β-carotene and LDs-triacylglycerol biosynthesis, enlarging storage space by engineering LDs related genes has minor promotion on β-carotene accumulation. Adding 2 mM OA significantly improved LDs-triacylglycerol metabolism and resulted in 36.4% increase in β-carotene content. The transcriptome analysis was adopted to mine OA-repressible promoters and IZH1 promoter was used to replace native ERG9 promoter to dynamically down-regulate ERG9 expression, which diverted the metabolic flux to β-carotene pathway and achieved additional 31.7% increase in β-carotene content without adversely affecting cell growth. By inducing an extra constitutive β-carotene synthesis pathway for further conversion precursor farnesol to β-carotene, the final strain produced 11.4 mg/g DCW and 142 mg/L of β-carotene, which is 107.3% and 49.5% increase respectively over the parent strain. This strategy can be applied in the overproduction of other heterogeneous FPP-derived hydrophobic compounds with similar synthesis and storage mechanisms in S. cerevisiae. The online version contains supplementary material available at 10.1186/s12934-021-01723-y.
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影响因子：
6.4
作者：
Chen Y;Xiao W;Wang Y;Liu H;Li X;Yuan Y
通讯作者：
Yuan Y
DOI：
10.1073/pnas.1301358110
发表时间：
2013-05-07
影响因子：
11.1
作者：
Doshi, Rupak;Tuan Nguyen;Chang, Geoffrey
通讯作者：
Chang, Geoffrey
影响因子：
2.9
作者：
Song, Linsheng
通讯作者：
Song, Linsheng
影响因子：
1.2
作者：
Kumar, Naresh M.;Thunuguntla, V. B. S. C.;Bondili, J. S.
通讯作者：
Bondili, J. S.
通过改善 ATP 供应和膜灵活性改造内源 ABC 转运蛋白可增强酿酒酵母中 β-胡萝卜素的分泌
影响因子：
6.3
作者：
Bu X;Lin JY;Cheng J;Yang D;Duan CQ;Koffas M;Yan GL
通讯作者：
Yan GL