Bu Ruihong, Yang Zhiheng, Li Zilong, Zhang Guojian, Wang Weishan
College of Medicine, Ocean University of China, Qingdao 266000, Shandong, China.
Institute of Microbiology, Chinese Academy of Sciences, Beijing 100101, China.
Sheng Wu Gong Cheng Xue Bao. 2023 Aug 25;39(8):3508-3519. doi: 10.13345/j.cjb.230015.
is a kind of Gram-positive facultative anaerobic bacteria. The fast growth rate under high temperature and less susceptibility to microbial contamination enable . to be a desirable producer of biofuels and high-value-added chemicals for the next-generation industrial biotechnology. However, compared with the classical model strain , the applications of . are hampered by its low transformation efficiency. This study aimed at obtaining competent cells with high transformation efficiency through inactivating restriction enzymes, adding cell membrane inhibitors and cell wall weakening agents. The results showed that the electro-transformation efficiency achieved 1.2×10 CFU/(μg DNA) by knocking out four genes encoding restriction enzymes. Adding a certain amount of tween 80, dl-threonine and glycine further increased the competent efficiency about 22.5, 44, and 334 times, respectively. The electro-transformation efficiency was enhanced to 4.6×10 CFU/(μg DNA) under the optimized conditions, laying a foundation for genetic manipulation and metabolic engineering of . .
是一种革兰氏阳性兼性厌氧菌。其在高温下生长速度快且对微生物污染的敏感性较低，使得它成为下一代工业生物技术中生物燃料和高附加值化学品的理想生产者。然而，与经典模式菌株相比，其应用受到低转化效率的阻碍。本研究旨在通过灭活限制酶、添加细胞膜抑制剂和细胞壁弱化剂来获得具有高转化效率的感受态细胞。结果表明，通过敲除四个编码限制酶的基因，电转化效率达到了1.2×10 CFU/（μg DNA）。添加一定量的吐温80、dl-苏氨酸和甘氨酸分别使感受态效率进一步提高了约22.5倍、44倍和334倍。在优化条件下，电转化效率提高到了4.6×10 CFU/（μg DNA），为其基因操作和代谢工程奠定了基础。
