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大肠杆菌中高产叶黄素的途径工程

Pathway engineering for high-yield production of lutein in Escherichia coli.

作者信息

Takemura Miho, Kubo Akiko, Watanabe Asuka, Sakuno Hanayo, Minobe Yuka, Sahara Takehiko, Murata Masahiro, Araki Michihiro, Harada Hisashi, Terada Yoshinobu, Yaoi Katsuro, Ohdan Kohji, Misawa Norihiko

机构信息

Research Institute for Bioresources and Biotechnology, Ishikawa Prefectural University, Nonoichi, Ishikawa, Japan.

Applied Research Laboratory, Ezaki Glico Co., Ltd., Osaka, Japan.

出版信息

Synth Biol (Oxf). 2021 May 15;6(1):ysab012. doi: 10.1093/synbio/ysab012. eCollection 2021.

Abstract

Lutein is an industrially important carotenoid pigment, which is essential for photoprotection and photosynthesis in plants. Lutein is crucial for maintaining human health due to its protective ability from ocular diseases. However, its pathway engineering research has scarcely been performed for microbial production using heterologous hosts, such as , since the engineering of multiple genes is required. These genes, which include tricky key carotenoid biosynthesis genes typically derived from plants, encode two sorts of cyclases (lycopene ε- and β-cyclase) and cytochrome P450 CYP97C. In this study, upstream genes effective for the increase in carotenoid amounts, such as isopentenyl diphosphate isomerase () gene, were integrated into the JM101 (DE3) genome. The most efficient set of the key genes ( and ) was selected from among the corresponding genes derived from various plant (or bacterial) species using that had accumulated carotenoid substrates. Furthermore, to optimize the production of lutein in , we introduced several sorts of plasmids that contained some of the multiple genes into the genome-inserted strain and compared lutein productivity. Finally, we achieved 11 mg/l as lutein yield using a mini jar. Here, the high-yield production of lutein was successfully performed using through approaches of pathway engineering. The findings obtained here should be a base reference for substantial lutein production with microorganisms in the future.

摘要

叶黄素是一种具有重要工业价值的类胡萝卜素色素,对植物的光保护和光合作用至关重要。叶黄素因其预防眼部疾病的能力而对维持人体健康至关重要。然而,由于需要对多个基因进行工程改造,其在诸如大肠杆菌等异源宿主微生物生产中的途径工程研究几乎尚未开展。这些基因包括通常源自植物的棘手的关键类胡萝卜素生物合成基因,编码两种环化酶(番茄红素ε-环化酶和β-环化酶)以及细胞色素P450 CYP97C。在本研究中,将对增加类胡萝卜素产量有效的上游基因,如异戊烯基二磷酸异构酶(idi)基因,整合到大肠杆菌JM101(DE3)基因组中。使用积累了类胡萝卜素底物的大肠杆菌,从源自各种植物(或细菌)物种的相应基因中筛选出最有效的一组关键基因(crtE和crtB)。此外,为了优化大肠杆菌中叶黄素的生产,我们将包含多个基因中的一些基因的几种质粒导入基因组插入菌株,并比较叶黄素的生产力。最后,我们使用小型摇瓶实现了11毫克/升的叶黄素产量。在此,通过途径工程方法成功地利用大肠杆菌实现了叶黄素的高产生产。此处获得的研究结果应成为未来利用微生物大量生产叶黄素的基础参考。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/4c78/8546607/7203afef4b38/ysab012f1.jpg

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