Enhanced production of β-carotene in recombinant Saccharomyces cerevisiae by inverse metabolic engineering with supplementation of unsaturated fatty acids
Enhanced production of β-carotene in recombinant Saccharomyces cerevisiae by inverse metabolic engineering with supplementation of unsaturated fatty acids
复制标题
通过补充不饱和脂肪酸的逆向代谢工程提高重组酿酒酵母中β-胡萝卜素的产量
DOI：
10.1016/j.procbio.2016.02.004
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发表时间：
2016-05-01
影响因子：
4.4
通讯作者：
Yan, Guoliang
中科院分区：
文献类型：
--
作者：
Sun, Yuxia;Sun, Liang;Yan, Guoliang
利用反代谢工程技术改进了重组酿酒酵母β -胡萝卜素的生物合成。甲羟戊酸途径基因转录谱测定结果显示，与亲本菌株相比，重组菌株诱导了HMG1、ERG8、ERG19和ERG20的转录，而HMG2、ERG12和IDI的表达无显著差异。重组酵母麦角甾醇和不饱和脂肪酸（UFAs）含量降低，参与其合成的ERG9和OLE1表达上调，表明重组酵母对麦角甾醇和不饱和脂肪酸的需求增加。单独过表达ERG19、ERG12和ERG20导致β -胡萝卜素含量增加11.0 ~ 32.2%，而过表达ERG8和IDI1分别减少22.9%和13.6%。相比之下，添加60 mg/l油酸和棕榈油酸分别使β -胡萝卜素含量增加83.7%和130.2%，细胞中UFA含量恢复。这些结果表明，甲羟戊酸途径中大多数基因的单一过表达并不是促进类胡萝卜素产生的有效途径。然而，细胞内UFA含量对类胡萝卜素的生物合成至关重要，通过外源补充UFA含量或加强其生物合成可能是提高重组酿酒酵母类胡萝卜素产量的一种有希望的策略。(C) 2016 Elsevier Ltd.版权所有。
beta-Carotene biosynthesis in recombinant Saccharomyces cerevisiae was improved using inverse metabolic engineering. Determination of the transcriptional profiles of mevalonate pathway genes showed that transcription of HMG1, ERG8, ERG19 and ERG20 were induced in recombinant strain compared with the parent strain, whereas the expression of HMG2, ERG12 and IDI showed no significant differences. The contents of ergosterol and unsaturated fatty acids (UFAs) were decreased in recombinant yeast, and ERG9 and OLE1 involved in their syntheses were upregulated, suggesting that an increased demand exists for ergosterol and UFAs in recombinant yeast. Single overexpression of ERG19, ERG12 and ERG20 led to 11.0-32.2% increments in beta-carotene content, but caused 22.9% and 13.6% decrements in overexpression of ERG8 and IDI1, respectively. In comparison, supplementation of 60 mg/l oleic acid and palmitoleic acid led to 83.7% and 130.2% increments of beta-carotene content, respectively, in parallel with recovery of UFA contents in cells. These results suggested that single overexpression of most genes in mevalonate pathway is not an effective approach for enhanced carotenoid production. However, intracellular UFA contents are important for carotenoid biosynthesis, and enriching UFA contents by exogenous supplementation or strengthening their biosynthesis might be a promising strategy to improve carotenoid production in recombinant S. cerevisiae. (C) 2016 Elsevier Ltd. All rights reserved.