文献类型：期刊文献

英文题名：Metabolic compartmentalization in yeast mitochondria: Burden and solution for squalene overproduction

作者：Zhu, Zhan-Tao[1];Du, Meng-Meng[1];Gao, Bei[1];Tao, Xin-Yi[1];Zhao, Ming[1];Ren, Yu-Hong[1];Wang, Feng-Qing[1];Wei, Dong-Zhi[1]

机构：[1]East China Univ Sci & Technol, Newworld Inst Biotechnol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2021

卷号：68

起止页码：232

外文期刊名：METABOLIC ENGINEERING

收录：;EI(收录号：20214411087471);WOS:【SCI-EXPANDED(收录号:WOS:000718926900002)】；

基金：Y This work was financially supported by the Natural Science Foun-dation of Shanghai (No.20ZR1415100) and the National Natural Science Foundation of China (No. 21776075) .

语种：英文

外文关键词：Saccharomyces cerevisiae; Mitochondrial engineering; Metabolic burden; Squalene; Dual engineering

摘要：Harnessing mitochondria is considered as a promising method for biosynthesis of terpenes due to the adequate supply of acetyl-CoA and redox equivalents in mitochondria. However, mitochondrial engineering often causes serious metabolic burden indicated by poor cell growth. Here, we systematically analyzed the metabolic burden caused by the compartmentalization of the MVA pathway in yeast mitochondria for squalene synthesis. The phosphorylated intermediates of the MVA pathway, especially mevalonate-5-P and mevalonate-5-PP, conferred serious toxicity within mitochondria, which significantly compromised its possible advantages for squalene synthesis and was difficult to be significantly improved by routine pathway optimization. These phosphorylated intermediates were converted into ATP analogues, which strongly inhibited ATP-related cell function, such as mitochondrial oxidative respiration. Fortunately, the introduction of a partial MVA pathway from acetyl-CoA to mevalonate in mitochondria as well as the augmentation of the synthesis of mevalonate in cytosol could significantly promote the growth of yeasts. Accordingly, a combinatorial strategy of cytoplasmic and mitochondrial engineering was proposed to alleviate the metabolic burden caused by the compartmentalized MVA pathway in mitochondria and improve cell growth. The strategy also displayed the superimposed effect of cytoplasmic engineering and mitochondrial engineering on squalene production. Through a two-stage fermentation process, the squalene titer reached 21.1 g/L with a specific squalene titer of 437.1 mg/g dcw, which was the highest at present. This provides new insight into the production of squalene and other terpenes in yeasts based on the advantages of mitochondrial engineering.