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

Development of a regulatable low-temperature protein expression system using the psychrotrophic bacterium, Shewanella livingstonensis Ac10, as the host
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使用耐冷细菌 Shewanella Livingstonensis Ac10 作为宿主开发可调节的低温蛋白表达系统

DOI:
10.1080/09168451.2019.1638754
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发表时间:
2019
期刊:
Bioscience, Biotechnology, and Biochemistry
影响因子:
--
通讯作者:
Tatsuo Kurihara
Tatsuo Kurihara
中科院分区:
--
文献类型:
--
作者:
Soichiro Kawai;Jun Kawamoto;Takuya Ogawa;Tatsuo Kurihara

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低温蛋白表达系统对耐热性蛋白的生产是有用的。我们之前开发了一个系统,使组成蛋白在低温下生产,使用精神营养细菌希瓦氏菌livingstonensisAc10作为宿主。为了提高该系统的实用性,在本研究中,我们将该细菌的tropoperon的抑制启动子引入该系统。在该启动子控制下,在18℃和4℃条件下产生ß-内酰胺酶时,在不含fl- trp的情况下,产量分别为75和33 mg/ l,在有fl- trp的情况下,产量分别下降了72%和77%。我们还发现,3-吲哚丙烯酸,大肠杆菌抑制因子的竞争性抑制剂,增加了报告基因的表达。这种可抑制的基因表达系统将有助于在低温下生产可调节的重组蛋白。
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 bacteriumShewanella livingstonensisAc10 as the host. To increase the utility of this system, in the present study, we introduced a repressible promoter of thetrpoperon 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 ofl-Trp, and the yields were decreased by 72% and 77%, respectively, in the presence ofl-Trp. We also found that 3-indoleacrylic acid, a competitive inhibitor of theEscherichia coli trprepressor, increased the expression of the reporter gene. This repressible gene expression system would be useful for regulatable recombinant protein production at low temperatures.
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