文献类型：期刊文献

英文题名：Metabolic engineering of β-carotene biosynthesis in Yarrowia lipolytica

作者：Zhang, Xin-Kai[1];Wang, Dan-Ni[1];Chen, Jun[1];Liu, Zhi-Jie[2];Wei, Liu-Jing[1];Hua, Qiang[1,3]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Hubei Univ Technol, Hubei Collaborat Innovat Ctr Ind Fermentat, Key Lab Fermentat Engn, Hubei Key Lab Ind Microbiol,Minist Educ, Wuhan 430068, Peoples R China;[3]Shanghai Collaborat Innovat Ctr Biomfg Technol, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2020

卷号：42

期号：6

起止页码：945

外文期刊名：BIOTECHNOLOGY LETTERS

收录：;EI(收录号：20200908213330);WOS:【SCI-EXPANDED(收录号:WOS:000516342500001)】；

基金：This study was financially supported by National Natural Science Foundation of China (21576089, 21776081), the 111 Project (B18022) and Research Program of State Key Laboratory of Bioreactor Engineering.

语种：英文

外文关键词：Yarrowia lipolytica; beta-carotene; CRISPR; Cas9 system; Metabolic engineering; Fermenter cultivation

摘要：Objective Carotenoids, as potent antioxidant compounds, have gained extensive attention, especially in human health. In this study, the combination of CRISPR/Cas9 integration strategy and fermenter cultivation was utilized to obtain efficient beta-carotene-producing Yarrowia lipolytica cell factories for potential industrial application. Results The introduction of the genes of Mucor circinelloides, encoding phytoene dehydrogenase (carB) and bifunctional phytoene synthase/lycopene cyclase (carRP), contributed to the heterologous production of beta-carotene in Y. lipolytica XK2. Furthermore, beta-carotene production was efficiently enhanced by increasing the copy numbers of the carB and carRP genes and overexpressing of GGS1, ERG13, and HMG, the genes related to the mevalonate (MVA) pathway. Thus, the optimized strain overexpressed a total of eight genes, including three copies of carRP, two copies of carB, and single copies of GGS1, HMG, and ERG13. As a consequence, strain Y. lipolytica XK19 accumulated approximately 408 mg/L beta-carotene in shake flask cultures, a twenty-four-fold increase compared to the parental strain Y. lipolytica XK2. Conclusions 4.5 g/L beta-carotene was obtained in a 5-L fermenter through a combination of genetic engineering and culture optimization, suggesting a great capacity and flexibility of Y. lipolytica in the production of carotenoids.