Boosting Geranyl Diphosphate Synthesis for Linalool Production in Engineered Yarrowia lipolytica

被引:3
作者
Taratynova, Maria O. [1 ]
Tikhonova, Ekaterina E. [1 ]
Fedyaeva, Iuliia M. [1 ]
Dementev, Dmitry A. [1 ]
Yuzbashev, Tigran V. [2 ]
Solovyev, Andrey I. [3 ]
Sineoky, Sergey P. [1 ]
Yuzbasheva, Evgeniya Y. [4 ,5 ]
机构
[1] NRC Kurchatov Inst, Kurchatov Genom Ctr, sq Acad Kurchatova 1, Moscow 123182, Russia
[2] Rothamsted Res, Plant Sci & Bioecon, Harpenden AL5 2JQ, West Common, England
[3] Russian Minist Hlth, Gamaleya Natl Res Ctr Epidemiol & Microbiol, Gamaleya St 18, Moscow 123098, Russia
[4] BioMediCan Inc, 40471 Encyclopedia Circle, Fremont, CA 94538 USA
[5] BioKai Inc, 40471 Encyclopedia Circle, Fremont, CA 94538 USA
关键词
Linalool; Geranyl diphosphate synthase; Yarrowia lipolytica; Metabolic engineering; Monoterpenoids; YEAST; OVERPRODUCTION; SUCROSE;
D O I
10.1007/s12010-023-04581-z
中图分类号
Q5 [生物化学]; Q7 [分子生物学];
学科分类号
071010 ; 081704 ;
摘要
Linalool is a pleasant-smelling monoterpenoid widely found in the essential oils of most flowers. Due to its biologically active properties, linalool has considerable commercial potential, especially in the food and perfume industries. In this study, the oleaginous yeast Yarrowia lipolytica was successfully engineered to produce linalool de novo. The (S)-linalool synthase (LIS) gene from Actinidia argute was overexpressed to convert geranyl diphosphate (GPP) into linalool. Flux was diverted from farnesyl diphosphate (FPP) synthesis to GPP by introducing a mutated copy of the native ERG20(F88W-N119W) gene, and CrGPPS gene from Catharanthus roseus on its own and as part of a fusion with LIS. Disruption of native diacylglycerol kinase enzyme, DGK1, by oligo-mediated CRISPR-Cas9 inactivation further increased linalool production. The resulting strain accumulated 109.6 mg/L of linalool during cultivation in shake flasks with sucrose as a carbon source. CrGPPS expression in Yarrowia lipolytica increased linalool accumulation more efficiently than the ERG20(F88W-N119W) expression, suggesting that the increase in linalool production was predominantly influenced by the level of GPP precursor supply.
引用
收藏
页码:1304 / 1315
页数:12
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