Kawai Soichiro, Kawamoto Jun, Ogawa Takuya, Kurihara Tatsuo
Laboratory of Molecular Microbial Science, Institute for Chemical Research, Kyoto University , Kyoto , Japan.
Biosci Biotechnol Biochem. 2019 Nov;83(11):2153-2162. doi: 10.1080/09168451.2019.1638754. Epub 2019 Jul 11.
A low-temperature protein expression system is useful for the production of thermolabile proteins. We previously developed a system that enables constitutive protein production at low temperatures, using the psychrotrophic bacterium Ac10 as the host. To increase the utility of this system, in the present study, we introduced a repressible promoter of the operon of this bacterium into the system. When ß-lactamase was produced under the control of this promoter at 18°C and 4°C, the yields were 75 and 33 mg/L-culture, respectively, in the absence of L-Trp, and the yields were decreased by 72% and 77%, respectively, in the presence of L-Trp. We also found that 3-indoleacrylic acid, a competitive inhibitor of the repressor, increased the expression of the reporter gene. This repressible gene expression system would be useful for regulatable recombinant protein production at low temperatures.
低温蛋白质表达系统对于生产热不稳定蛋白质很有用。我们之前开发了一种系统，该系统利用嗜冷细菌Ac10作为宿主，能够在低温下组成型生产蛋白质。为了提高该系统的实用性，在本研究中，我们将这种细菌操纵子的可阻遏启动子引入该系统。当β-内酰胺酶在该启动子的控制下于18°C和4°C生产时，在不存在L-色氨酸的情况下，产量分别为75和33 mg/L培养物，而在存在L-色氨酸的情况下，产量分别降低了72%和77%。我们还发现，阻遏物的竞争性抑制剂3-吲哚丙烯酸增加了报告基因的表达。这种可阻遏基因表达系统将有助于在低温下进行可调节的重组蛋白生产。
