代谢工程改造酿酒酵母发酵生产β-胡萝卜素  

Metabolic engineering modified Saccharomyces cerevisiae to produceβ-carotene by fermentation

作　　者：李渊涛 石贵阳 张梁[1,2] 丁重阳 周景文[1,2] 李由然 徐沙[1,2] LI Yuantao;SHI Guiyang;ZHANG Liang;DING Chongyang;ZHOU Jingwen;LI Youran;XU Sha(National Engineering Research Center for Cereal Fermentation and Food Biomanufacturing,Jiangnan University,Wuxi 214122,China;School of Biotechnology,Jiangnan University,Wuxi 214122,China)

机构地区：[1]江南大学,粮食发酵与食品生物制造国家工程研究中心,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与发酵工业》2024年第14期26-34,共9页Food and Fermentation Industries

基　　金：国家自然科学基金(22278188)。

摘　　要：β-胡萝卜素属于多烯烃类,是水果和蔬菜中含量最高的类胡萝卜素,能够预防并减轻心脏病,白内障和癌症等疾病,市场需求量较大,利用生物合成法生产β-胡萝卜素具有较好的应用前景。胞质乙酰辅酶A是β-胡萝卜素合成的重要前体,如何提高胞质乙酰辅酶A的含量是高效合成β-胡萝卜素的关键。该研究利用米曲霉(Aspergillus oryzae)来源的ATP-柠檬酸裂解酶基因(ATP-citrate lyase,ACL)转化出发菌株ZS20,同时过表达酿酒酵母(Saccharomyces cerevisiae)自身CAB 1基因,从而增加胞质乙酰辅酶A含量,β-胡萝卜素产量显著提高,菌株L5的摇瓶发酵产量达到333.3 mg/L,是出发菌株的2.1倍。在此基础上,菌株L5回补缺陷的腺嘌呤、赖氨酸、亮氨酸和色氨酸合成关键基因得到菌株L11,β-胡萝卜素摇瓶发酵产量为378.1 mg/L。经发酵罐发酵优化后,产量提高到1152.7 mg/L。β-carotene,a member of the polyolefin family,is the predominant carotenoid found in fruits and vegetables.It possesses the ability to prevent and alleviate various diseases,such as heart disease,cataracts,and cancer.As a result,it is highly sought after in the market due to its potential health benefits.The potential forβ-carotene production through biosynthesis holds promising prospects.Significantly,cytoplasmic acetyl coenzyme A is a crucial precursor forβ-carotene synthesis,highlighting the importance of increasing its content for efficientβ-carotene production.In this study,the ATP-citrate lyase(ACL)from Aspergillus oryzae was introduced into the starter strain Saccharomyces cerevisiae ZS20,in which the CAB1 gene was also overexpressed,to increase the cytoplasmic acetyl-coenzyme A content.As a result,theβ-carotene production significantly increased,reaching 333.3 mg/L in the flask fermentation of strain L5,which was a 2.1-fold increase compared to the starter strain.To further enhance the yield ofβ-carotene production,strain L5 was utilized to complement the deficient key genes responsible for adenine,lysine,leucine,and tryptophan synthesis,resulting in the development of strain L11.Initially,strain L11 demonstrated aβ-carotene yield of 378.1 mg/L when cultivated in shaking flasks.Subsequently,through additional optimization of the fermentation process carried out in the fermenter,the yield was significantly enhanced to 1152.7 mg/L.

关 键 词：酿酒酵母 Β-胡萝卜素 乙酰辅酶A ATP-柠檬酸裂解酶 氨基酸 

分 类 号：TQ920.6[轻工技术与工程—发酵工程]