Metabolic engineering of oleaginous yeast Yarrowia lipolytica for limonene overproductionReportar como inadecuado

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Biotechnology for Biofuels

, 9:214

First Online: 11 October 2016Received: 18 June 2016Accepted: 27 September 2016


BackgroundLimonene, 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.

ResultsTo 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.

ConclusionsThis 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.

KeywordsLimonene Yarrowia lipolytica Neryl diphosphate synthase 1 Limonene synthase AbbreviationsDMAPPdimethylallyl pyrophosphate

IPPisopentenyl pyrophosphate

GPPgeranyl diphosphate

NPPneryl diphosphate


NDPS1neryl diphosphate synthase

LSlimonene synthase

Electronic supplementary materialThe online version of this article doi:10.1186-s13068-016-0626-7 contains supplementary material, which is available to authorized users.

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Autor: Xuan Cao - Yu-Bei Lv - Jun Chen - Tadayuki Imanaka - Liu-Jing Wei - Qiang Hua


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