食品与发酵工业2026，Vol.52Issue(18)：1-11,11.DOI:10.13995/j.cnki.11-1802/ts.045299
通过常压室温等离子体诱变和基因组复制工程辅助连续进化技术提升酿酒酵母琥珀酸产量及其发酵优化
Enhanced succinic acid production in Saccharomyces cerevisiae via combinational use of atmospheric mutagenesis and room temperature plasma and genome replication engineering assisted continuous evolution techniques and fermentation optimization
摘要
Abstract
The kokumi taste of soy sauce is closely related to its quality.Succinic acid is one of the key compounds enhancing this at-tribute.This study aimed to improve the yield of succinic acid by a yeast strain via non-genetically engineered microbial breeding technolo-gies and to optimize the fermentation process,thereby providing technical support for enhancing the kokumi quality of soy sauce.Firstly,the physicochemical properties and sensory characteristics of commercial soy sauce were analyzed and a significant positive correlation be-tween succinic acid concentration and kokumi intensity was observed.Then,a combinational strategy of atmospheric and room temperature plasma(ARTP)mutagenesis and genome replication engineering assisted continuous evolution(GREACE)was applied to improve the succinic acid yield using Saccharomyces cerevisiae LA-FR as start strain.Through ARTP mutagenesis,the mutant strain LA-FR-100 was obtained and further subjected to GREACE evolution combined with high-salinity stresses.A genetically stable evolved strain LA-FR-100-15 with a succinic acid yield of 620.12 mg/L was obtained,which was 106％higher than that of start strain.Finally,the fermentation conditions for succinic acid production by the evolved strain were optimized.The optimal medium composition was as follows,glucose 80 g/L,yeast extract 13 g/L,corn steep liquor 7 g/L,K2HPO4·3H2O 3 g/L,MgSO4·7H2O 1.5 g/L and ZnSO4·7H2O 0.5 g/L.The optimal fermentation parameters were 26 ℃,initial pH 6.5,and 9％inoculum.Under the optimal fermentation condition,the yield of succinic acid by LA-FR-100-15 strain reached 1.54 g/L,which was 148.7％higher than that before optimization.Through fed-batch fer-mentation in 5 L fermenter,the final succinic acid yield reached 9.6 g/L.The results obtained in this study delivered a non-genetically engineered microbial solution and an optimized process to enhance the kokumi of soy sauce.关键词
酿酒酵母/琥珀酸/常压室温等离子体/基因组复制工程辅助连续进化/发酵优化/补料发酵Key words
Saccharomyces cerevisiae/succinic acid/atmospheric and room temperature plasma/genome replication engineering assisted continuous evolution/fermentation optimization/fed-batch fermentation引用本文复制引用
王雅欣,尚贝贝,许睿琦,钮成拓,郑飞云,王金晶,刘春凤,李崎..通过常压室温等离子体诱变和基因组复制工程辅助连续进化技术提升酿酒酵母琥珀酸产量及其发酵优化[J].食品与发酵工业,2026,52(18):1-11,11.基金项目
国家自然科学基金面上项目(32272282) （32272282）
