As an important aromatic alcohols with favorable flavor and superior property, 2-phenylethanol has been widely used in food, medicine, and chemical industry. In this study, the deletion of phenylpyruvate decarboxylase gene PDC5 was found to improve the synthesis of 2-phenylethanol, and related metabolic strategy of the Ehrlich pathway was rational reconstructed in Saccharomyces cerevisiae for further product-enhancement. Strain RM22 (pdc5△) was obtained using CRISPR/Cas9, and improved 2-phenylethanol production was observed in the optimized culture conditions. Additionally, by expressing aromatic transaminase Aro8p and Aro9p respectively, we found that overexpressing of Aro8p in aro9-deleted strain was an efficient strategy to increase 2-phenylethanol production. To further improve the production of 2-phenylethanol, metabolic engineered strain RM-59810 (pdc5△aro9△PTPI-ARO8-TTT PTPI-ARO10-TTT) was constructed, resulting in 3.85 g/L of 2-phenylethanol from 6.7 g/L phenylalanine after 120 h, and the molar conversion rate was 0.67 mol/mol, which was 1.33-fold than that of the control strain, with a yield of 0.5 g/g L-phenylalanine. In this study, a new efficient strategy was provided to improve 2-phenylethanol production in S. cerevisiae, and also provided a primary foundation for studying the possible regulatory effects of pyruvate decarboxylase Pdc5p.
ZHU Linghuan
,
XU Sha
,
LI Youran
,
ZHANG Liang
,
SHI Guiyang
. Improvement of 2-phenylethanol production by deleting gene PDC5 and related metabolic strategies in Saccharomyces cerevisiae[J]. Food and Fermentation Industries, 2021
, 47(16)
: 22
-30
.
DOI: 10.13995/j.cnki.11-1802/ts.027019
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