Exchanging the order of carotenogenic genes linked by porcine teschovirus-1 2A peptide enable to optimize carotenoid metabolic pathway in Saccharomyces cerevisiae†,RSC Advances

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Exchanging the order of carotenogenic genes linked by porcine teschovirus-1 2A peptide enable to optimize carotenoid metabolic pathway in Saccharomyces cerevisiae†
RSC Advances ( IF 3.9 ) Pub Date : 2018-10-12 00:00:00 , DOI: 10.1039/c8ra06510a
Xiang Jiao _{1,

2} , Wenyi Sun ₁ , Yue Zhang ₁ , Xiangjian Liu ₁ , Qi Zhang ₁ , Qian Wang ₁ , Sufang Zhang _{1,

3} , Zongbao Kent Zhao _{1,

3}

Affiliation

The yeast Saccharomyces cerevisiae serves as a promising host for the production of a wide range of chemical compounds and fuels. Currently, simultaneous expression of several genes could be achieved via the use of 2A viral peptides, yet detailed characterizations to assess the discrepancy of different orders of genes linked by 2A peptides are rarely sufficient. In this study, we investigated the effects of the order of genes linked by porcine teschovirus-1 2A (P2A) peptide on the metabolic pathway in S. cerevisiae. A heterologous carotenoid biosynthetic system involving nine kinds of polycistronic expression of codon-optimized carotenogenic genes GGPPS, CARB and CARRP from Blakeslea trispora was introduced into S. cerevisiae. The order of genes in the polycistronic segment was exchanged; β-carotene production by engineered yeasts was significantly different. The highest β-carotene yield was achieved in transformants carrying the plasmid, with CARB as the first gene in the polycistronic construct regardless of the location of GGPPS, CARRP. In addition, we found that β-carotene production was coupled with the growth in engineered strain with the highest β-carotene content during the shake flask fermentation and fed-batch fermentation. A novel microbial heterologous carotenoid production system was established by optimizing the order of genes linked by P2A peptide sequences in a polycistronic expression construct. The observation of the importance of the order in a polycistronic construct may be used to increase yields in other P2A peptide-containing expression systems.

中文翻译：

交换猪teschovirus-1 2A肽连接的类胡萝卜素基因顺序优化酿酒酵母类胡萝卜素代谢途径†

酵母酿酒酵母是生产多种化合物和燃料的有前途的宿主。目前，可以通过使用 2A 病毒肽实现多个基因的同时表达，但评估由 2A 肽连接的不同基因顺序的差异的详细表征很少足够。在这项研究中，我们研究了由猪 teschovirus-1 2A (P2A) 肽连接的基因顺序对酿酒酵母代谢途径的影响。一种异源类胡萝卜素生物合成系统，涉及密码子优化的类胡萝卜素基因GGPPS、CARB和CARRP的9 种多顺反子表达Blakeslea trispora被引入酿酒酵母。多顺反子片段中的基因顺序被交换；工程酵母产生的β-胡萝卜素显着不同。无论GGPPS、CARRP的位置如何，在携带质粒的转化子中实现了最高的 β-胡萝卜素产量，其中CARB作为多顺反子构建体中的第一个基因. 此外，我们发现在摇瓶发酵和补料分批发酵过程中，β-胡萝卜素的产生与β-胡萝卜素含量最高的工程菌株的生长有关。通过优化多顺反子表达构建体中P2A肽序列连接的基因顺序，建立了一种新型微生物异源类胡萝卜素生产系统。观察多顺反子构建体中顺序的重要性可用于增加其他含 P2A 肽的表达系统的产量。

更新日期：2018-10-12

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