Development of a regulatable low-temperature protein expression system using the psychrotrophic bacterium, Shewanella livingstonensis Ac10, as the host

• Published in: Bioscience, biotechnology, and biochemistry biotechnology, and biochemistry, 2019-11, Vol.83 (11), p.2153-2162
• Main Authors: Kawai, Soichiro, Kawamoto, Jun, Ogawa, Takuya, Kurihara, Tatsuo
• Format: Article
• Language: English
• Subjects: Cold Temperature; Gene Expression; Genetic Engineering - methods; heterologous protein production; Operon - genetics; Promoter Regions, Genetic - genetics; Recombinant Proteins - biosynthesis; Recombinant Proteins - genetics; regulatable gene expression; Shewanella; Shewanella - genetics; Trp promoter
• ISSN: 0916-8451; 1347-6947
• DOI: 10.1080/09168451.2019.1638754

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 Shewanella livingstonensis Ac10 as the host. To increase the utility of this system, in the present study, we introduced a repressible promoter of the trp 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 Escherichia coli trp repressor, increased the expression of the reporter gene. This repressible gene expression system would be useful for regulatable recombinant protein production at low temperatures. A regulatable low-temperature protein expression system was constructed by using a cold-adapted bacterium as the host and its trp operon promoter.