文献类型：期刊文献

英文题名：Metabolic engineering of oleaginous yeast Yarrowia lipolytica for limonene overproduction

作者：Cao, Xuan[1];Lv, Yu-Bei[1];Chen, Jun[1];Imanaka, Tadayuki[1];Wei, Liu-Jing[1];Hua, Qiang[1,2]

机构：[1]East China Univ Sci & Technol, State Key Lab Bioreactor Engn, 130 Meilong Rd, Shanghai 200237, Peoples R China;[2]Shanghai Collaborat Innovat Ctr Biomfg Technol, 130 Meilong Rd, Shanghai 200237, Peoples R China

年份：2016

卷号：9

期号：1

外文期刊名：BIOTECHNOLOGY FOR BIOFUELS

收录：;EI(收录号：20164202919216);WOS:【SCI-EXPANDED(收录号:WOS:000385580000001)】；

基金：This work was financially supported by National Basic Research Program of China (973 Program) (2012CB721101), and National Natural Science Foundation of China (21576089).

语种：英文

外文关键词：Limonene; Yarrowia lipolytica; Neryl diphosphate synthase 1; Limonene synthase

摘要：Background: Limonene, a monocyclic monoterpene, is known for its using as an important precursor of many flavoring, pharmaceutical, and biodiesel products. Currently, d-limonene has been produced via fractionation from essential oils or as a byproduct of orange juice production, however, considering the increasing need for limonene and a certain amount of pesticides may exist in the limonene obtained from the citrus industry, some other methods should be explored to produce limonene. Results: To construct the limonene synthetic pathway in Yarrowia lipolytica, two genes encoding neryl diphosphate synthase 1 (NDPS1) and limonene synthase (LS) were codon-optimized and heterologously expressed in Y. lipolytica. Furthermore, to maximize limonene production, several genes involved in the MVA pathway were overexpressed, either in different copies of the same gene or in combination. Finally with the optimized pyruvic acid and dodecane concentration in flask culture, a maximum limonene titer and content of 23.56 mg/L and 1.36 mg/g DCW were achieved in the final engineered strain Po1f-LN-051, showing approximately 226-fold increase compared with the initial yield 0.006 mg/g DCW. Conclusions: This is the first report on limonene biosynthesis in oleaginous yeast Y. lipolytica by heterologous expression of codon-optimized tLS and tNDPS1 genes. To our knowledge, the limonene production 23.56 mg/L, is the highest limonene production level reported in yeast. In short, we demonstrate that Y. lipolytica provides a compelling platform for the overproduction of limonene derivatives, and even other monoterpenes.