posted on 2021-01-11, 16:38authored byPingping Zhou, Yi Du, Xin Fang, Nannan Xu, Chunlei Yue, Lidan Ye
Linalool,
as a fragrant monoterpene, is an important feedstock
for food, pharmaceuticals, and cosmetics industries. Although our
previous study had significantly increased linalool production by
the directed evolution of linalool synthase and overexpression of
the whole mevalonate pathway genes, the engineered yeast strain suffered
from dramatically reduced biomass. Herein, a stress-free linalool-producing
yeast cell factory was constructed by the combinational regulation
of linalool synthase and farnesyl diphosphate synthase instead of
multienzyme overexpression. First, the expression level of linalool
synthase was successfully enhanced by introducing a N-terminal SKIK
tag, which improved linalool production by 3.3-fold. Subsequently,
the modular assembly of linalool synthase and dominant negative farnesyl
diphosphate synthase via short peptide tags efficiently converted
geranyl pyrophosphate to linalool. Additional downregulation of the
native farnesyl diphosphate synthase led to the highest reported linalool
production (80.9 mg/L) in yeast. This combinatorial modulation strategy
may also be applied to the production of other high-value monoterpenes.