产5-氨基乙酰丙酸酿酒酵母工程菌株的构建  

Construction of Recombinant Saccharomyces cerevisiae for Production of 5-Aminolevulinic Acid

作　　者：张俊丽[1,2] 康振[1,2] 钱晟东 邱玲[1,2] 陈坚[1,2] 堵国成[1,2] ZHANG Junl;KANG Zhen;QIAN Shengdong;QIU Ling;CHEN Jian;DU Guocheng(Key Laboratory of Industrial Biotechnology. Ministry of Education, Jiangnan University, Wuxi 214122, China;School of Biotechnology, Jiangnan University, Wuxi 214122, China)

机构地区：[1]江南大学工业生物技术教育部重点实验室,江苏无锡214122 [2]江南大学生物工程学院,江苏无锡214122

出　　处：《食品与生物技术学报》2018年第3期232-239,共8页Journal of Food Science and Biotechnology

基　　金：国家自然科学基金项目(31200020);国家973计划项目(2013CB733602;2014CB745103);长江学者和创新团队发展计划项目(IRT1135)

摘　　要：5-氨基乙酰丙酸(5-aminolevulinic acid,ALA)是一种天然存在的非蛋白质类氨基酸,在医学和农业领域应用广泛。为构建生产ALA的酿酒酵母细胞工厂,作者首先在酿酒酵母中分别表达了来源于酿酒酵母和类球红细菌中编码ALA生物合成C4途径ALA合酶的hem1和hemA基因,同时又表达了来源于大肠杆菌编码C5途径关键酶谷氨酰-tRNA还原酶和谷氨醛氨基转移酶的基因hemA和hemL。结果表明,表达酿酒酵母自身来源的hem1基因更利于ALA的合成,ALA产量为327.6 mg/L。在此基础上将C4和C5途径的基因hem1、hemA和hemL在酿酒酵母中过量表达,在添加前体物质甘氨酸和琥珀酸的条件下,ALA产量为525.8 mg/L,实现了在酿酒酵母中合成ALA。5-aminolevulinic acid （ALA）,a naturally occurring non-protein amino acid,has been widely applied in the industry of medicine and agriculture. To construct a recombinant Saccharomyces cerevisiae for production of ALA,genes hem1 cloned from S. cerevisiae and hemA cloned from Rhodobacter spheroides 2.4.1 encoded ALA synthase of C4 pathway was overexpressed,respectively. Meanwhile,glutamyl-tRNA reductase （encoded by hemA ） and glutamate-l-selnialdehyde aminotransferase （encoded by hemL）,the key enzymes of C5 pathway from Escherichia coli were also overexpressed. The result demonstrated that overexpression ofheml was more beneficial to the production of ALA, 327.6 mg/L. Moreover,genes heml,hemA and/wmL were co-overexpressed,resulting recombinant strain IN-PYIAL. The production of ALA was 525.8 mg/L with addition of precursor glycine and succinic acid. This realized the biosynthsis of ALA in S. eerevisiae.

关 键 词：5-氨基乙酰丙酸 C4途径 C5途径 5-氨基乙酰丙酸合酶 酿酒酵母 

分 类 号：TQ922.9[轻工技术与工程—发酵工程]