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通过过表达番茄红素 ε 环化酶基因提高莱茵衣藻叶黄素的产量。

Enhanced Lutein Production in Chlamydomonas reinhardtii by Overexpression of the Lycopene Epsilon Cyclase Gene.

机构信息

Graduate School of Agriculture, Kyoto University, Kitashirakawa Oiwake-cho, Sakyo-ku, Kyoto, 606-8502, Japan.

出版信息

Appl Biochem Biotechnol. 2021 Jun;193(6):1967-1978. doi: 10.1007/s12010-021-03524-w. Epub 2021 Feb 2.

Abstract

Chlamydomonas reinhardtii is a well-established microalgal model species with a shorter doubling time, which is a promising natural source for the efficient production of high-value carotenoids. In the microalgal carotenoid biosynthetic pathway, lycopene is converted either into β-carotene by lycopene β-cyclase or into α-carotene by lycopene ε-cyclase (LCYE) and lycopene β-cyclase. In this study, we overexpressed the LCYE gene in C. reinhardtii to estimate its effect on lycopene metabolism and lutein production. Chlamydomonas transformants (CrLCYE#L1, #L5, and #L6) produced significantly increased amounts of lutein per culture (up to 2.6-fold) without a decrease in cell yields. Likewise, the expression levels of LCYE gene in transformants showed a significant increase compared with that of the wild-type strain. These results suggest that LCYE overexpression enhances the conversion of lycopene to α-carotene, which in turn improves lutein productivity. Interestingly, their β-carotene productivity appeared to increase slightly rather than decrease. Considering that the inhibition of the lycopene cyclization steps often induces higher expression in genes upstream of metabolic branches, this result implies that the redirection from β-carotene to α-carotene by LCYE overexpression might also enhance upstream gene expression, thereby leading to auxiliary β-carotene production.

摘要

莱茵衣藻是一种经过充分验证的微藻模式生物,其倍增时间较短,是高效生产高价值类胡萝卜素的有前途的天然来源。在微藻类胡萝卜素生物合成途径中,番茄红素要么通过番茄红素β-环化酶转化为β-胡萝卜素,要么通过番茄红素ε-环化酶(LCYE)和番茄红素β-环化酶转化为α-胡萝卜素。在本研究中,我们在莱茵衣藻中过表达了 LCYE 基因,以评估其对番茄红素代谢和叶黄素生产的影响。衣藻转化体(CrLCYE#L1、#L5 和#L6)产生的叶黄素量每培养物显著增加(高达 2.6 倍),而细胞产量没有下降。同样,与野生型菌株相比,转化体中 LCYE 基因的表达水平也显著增加。这些结果表明,LCYE 的过表达增强了番茄红素向α-胡萝卜素的转化,从而提高了叶黄素的生产力。有趣的是,它们的β-胡萝卜素生产力似乎略有增加而不是减少。考虑到番茄红素环化步骤的抑制通常会导致代谢分支上游基因的更高表达,这一结果意味着 LCYE 过表达将β-胡萝卜素重新定向为α-胡萝卜素也可能增强上游基因表达,从而导致辅助β-胡萝卜素的产生。

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