Saccharomyces cerevisiae has been used for producing natural 2-phenylethanol (2-PE), but it is limited by low titer and weak tolerance. Three S. cerevisiae strains with good traits were screened, LSC-1 has a 2-PE yield of 3.41 g/L, NGER could tolerate 3.60 g/L of 2-PE, showing great stress tolerance, and S.C-1 showed thermotolerance and could grow at 41 ℃. A fusant strain RH2-16 was obtained through two rounds of protoplast fusion with these three strains as parents. The RH2-16 showed increased 2-PE tolerance by 20% and produced 4.31 g/L of 2-PE by fermentation conversion for 36 h with L-phenylalanine as substrate, an increase of 26.4% and 38.1% when compared to the parental strain LSC-1 and the industrial strain CWY132, respectively. With CRISPR/Cas9 system, mutated Ras-specific guanine nucleotide exchange factor CDC25 (W1416C) was constructed in RH2-16 and the yield of 2-PE increased by 8%. Overexpression of the key enzyme genes ARO8, ARO10, and ADH2 in the Ehrlich pathway in RH2-16 did not increase 2-PE production. Our study provides an effective strategy for screening and breeding of excellent strains of S. cerevisiae producing 2-PE and other natural products.
LIN Lucheng
,
XU Zhiwei
,
ZHANG Jianze
,
SHAN Yudong
,
XIAO Feng
,
XU Hao
,
LI Qinping
,
YI Puhong
,
WANG Kun
,
ZHU Tingheng
. Protoplast fusion combined with gene editing technology significantly improves the ability of Saccharomyces cerevisiae to produce 2-phenylethanol[J]. Food and Fermentation Industries, 2023
, 49(5)
: 18
-24
.
DOI: 10.13995/j.cnki.11-1802/ts.031898
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