Wang Li, Wang Yang, Li Jianghua, DU Guocheng, Kang Zhen
Key Laboratory of Carbohydrate Chemistry and Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
Key Laboratory of Industrial Biotechnology, Ministry of Education, School of Biotechnology, Jiangnan University, Wuxi 214122, Jiangsu, China.
Sheng Wu Gong Cheng Xue Bao. 2022 Feb 25;38(2):796-806. doi: 10.13345/j.cjb.210166.
Ergothioneine (ERG) is a natural antioxidant that has been widely used in the fields of food, medicine and cosmetics. Compared with traditional plant extraction and chemical synthesis approaches, microbial synthesis of ergothioneine has many advantages, such as the short production cycle and low cost, and thus has attracted intensive attention. In order to engineer an ergothioneine high-yielding strain, the ergothioneine synthesis gene cluster from and from were introduced into BL21(DE3) to generate a strain E1-A1 harboring the ergothioneine biosynthesis pathway. As a result, (95.58±3.2) mg/L ergothioneine was produced in flask cultures. To further increase ergothioneine yield, the relevant enzymes for biosynthesis of histidine, methionine, and cysteine, the three precursor amino acids of ergothioneine, were overexpressed. Individual overexpression of and resulted in an ergothioneine titer of (134.83±4.22) mg/L and (130.26±3.34) mg/L, respectively, while co-overexpression of and increased the production of ergothioneine to (144.97±5.40) mg/L. Eventually, by adopting a fed-batch fermentation strategy in 3 L fermenter, the optimized strain E1-A1- produced 548.75 mg/L and 710.53 mg/L ergothioneine in glucose inorganic salt medium and rich medium, respectively.
麦角硫因（ERG）是一种天然抗氧化剂，已广泛应用于食品、医药和化妆品领域。与传统的植物提取和化学合成方法相比，微生物合成麦角硫因具有许多优点，如生产周期短、成本低，因此受到了广泛关注。为了构建一株麦角硫因高产菌株，将来自[具体来源1]和[具体来源2]的麦角硫因合成基因簇导入大肠杆菌BL21(DE3)，构建了一株具有麦角硫因生物合成途径的菌株E1-A1。结果，摇瓶培养中产生了（95.58±3.2）mg/L的麦角硫因。为了进一步提高麦角硫因产量，过表达了麦角硫因的三种前体氨基酸（组氨酸、蛋氨酸和半胱氨酸）生物合成的相关酶。单独过表达[具体基因1]和[具体基因2]时，麦角硫因产量分别达到（134.83±4.22）mg/L和（130.26±3.34）mg/L，而同时过表达[具体基因1]和[具体基因2]时，麦角硫因产量提高到（144.97±5.40）mg/L。最终，通过在3 L发酵罐中采用分批补料发酵策略，优化后的菌株E1-A1-在葡萄糖无机盐培养基和丰富培养基中分别产生了548.75 mg/L和710.53 mg/L的麦角硫因。
