摘要
咖啡酸是一种天然酚酸类化合物,具有抗氧化、抗肿瘤、抗炎等多种生物学活性,咖啡酸也是迷迭香酸、绿原酸和咖啡酸苯乙酯等高附加值化合物合成的重要前体,在医药、食品和化妆品等行业具有较大的应用价值。为实现咖啡酸绿色可持续生产,在前期构建的产咖啡酸酿酒酵母菌株中,敲除苯丙氨酸途径中的预苯酸脱水酶基因(PHA2),将咖啡酸产量提高42%。而敲除色氨酸途径中邻氨基苯甲酸合成酶基因(TRP2),对咖啡酸积累影响不大。之后,回补了URA3、HIS3、MET15营养标记基因,然后将工程菌株在5 L的发酵罐中进行了补料分批发酵,使得咖啡酸产量达到了9.3 g/L,是目前文献中报道微生物发酵生产咖啡酸的最高水平,该研究为咖啡酸的绿色生产以及其衍生物的高效微生物合成奠定了基础。
Caffeic acid is a natural phenolic acid with various pharmaceutical properties,including antioxidant,anti-tumor and anti-inflammatory effects.It is also an important precursor for biosynthesis of other valuable compounds such as rosmarinic acid,chlorogenic acid and phenethyl caffeate.Therefore,caffeic acid has significant value in pharmaceutical,food,cosmetic and other industries.To achieve green and sustainable production of caffeic acid,the prephenate dehydratase gene(PHA2),which is involved in the biosynthesis of phenylalanine,was knocked out in a previously constructed caffeic acid produced Saccharomyces cerevisiae,resulting in a 42%improvement in caffeic acid production.Additionally,the anthranilate synthase gene(TRP2),which is responsible for tryptophan biosynthesis,was deleted.However,this had little effect on coffee acid production.The engineered strain was further complemented with auxotrophic marker URA3,HIS3,MET15.In a subsequent fed-batch fermentation process conducted in a 5 L bioreactor,the engineered strain achieved a caffeic acid production of 9.3 g/L,this is currently the highest reported titer of caffeic acid produced by engineered microbial cells.This study provides a foundation for the green production of caffeic acid and its derivatives.
作者
袁豆豆
周秀琪
庞雪晴
杜家艳
周萍萍
YUAN Doudou;ZHOU Xiuqi;PANG Xueqing;DU Jiayan;ZHOU Pingping(College of Bioscience and Biotechnology,Yangzhou University,Yangzhou 225009,China)
出处
《食品与发酵工业》
CAS
CSCD
北大核心
2024年第19期17-24,共8页
Food and Fermentation Industries
基金
国家自然科学基金项目(32371480,32001032)
扬州大学“高端人才支持计划”资助项目
江苏省大学生创新创业训练计划项目(202211117105Y)。
关键词
酿酒酵母
咖啡酸
预苯酸脱水酶
邻氨基苯甲酸合成酶
生物合成
Saccharomyces cerevisiae
caffeic acid
prephenate dehydratase
anthranilate synthase
biosynthesis
