Wang Jianrong, Zhang Tianyu, Li Yangyuan, Li Lilang, Wang Yong, Yang Bo, Wang Yonghua
1School of Bioscience and Bioengineering, South China University of Technology, Guangzhou, 510006 People's Republic of China.
Guangdong VTR Bio-Tech Co., Ltd, Zhuhai, 519060 People's Republic of China.
3 Biotech. 2019 Jan;9(1):33. doi: 10.1007/s13205-018-1531-5. Epub 2019 Jan 5.
In this study, 15 methanol-inducible and 9 constitutive promoters were used to drive the expression of lipase (TDL) in . Of the 15 methanol-inducible promoters, formaldehyde dehydrogenase promoter (P) showed the highest efficiency in driving lipase production, followed by alcohol oxidase 1 (P) and dihydroxyacetone synthase (P) promoters. The maximum lipase activity of transformants with P, P and P promoters in 5-l bioreactor was 27,076, 24,159 and 22,342 U/ml, respectively. For the nine constitutive promoters, glycosyl phosphatidyl inositol-anchored protein promoter (P) produced the highest amount of lipases in a medium containing glucose or glycerol as the only carbon source, followed by mitochondrial alcohol dehydrogenase isozyme (P) and glyceraldehyde-3-phosphate dehydrogenase (P) promoters. The maximum lipase yields in 5-l bioreactors under the control of P, P and P promoters were 17,353, 15,046 and 14,276 U/ml, respectively. The result of this study not only identifies a few highly efficient promoters for the heterologous expression of TDL in , but also casts some insight into the optimization of protein production in heterologous systems.
在本研究中，使用了15个甲醇诱导型启动子和9个组成型启动子来驱动脂肪酶（TDL）在……中的表达。在15个甲醇诱导型启动子中，甲醛脱氢酶启动子（P）在驱动脂肪酶产生方面表现出最高效率，其次是乙醇氧化酶1（P）和二羟基丙酮合酶（P）启动子。在5升生物反应器中，具有P、P和P启动子的转化体的最大脂肪酶活性分别为27,076、24,159和22,342 U/ml。对于9个组成型启动子，糖基磷脂酰肌醇锚定蛋白启动子（P）在以葡萄糖或甘油作为唯一碳源的培养基中产生的脂肪酶量最高，其次是线粒体乙醇脱氢酶同工酶（P）和甘油醛-3-磷酸脱氢酶（P）启动子。在P、P和P启动子控制下，5升生物反应器中的最大脂肪酶产量分别为17,353、15,046和14,276 U/ml。本研究结果不仅为TDL在……中的异源表达鉴定了一些高效启动子，也为异源系统中蛋白质生产的优化提供了一些见解。
