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中国生物工程杂志

China Biotechnology
China Biotechnology  2016, Vol. 36 Issue (3): 61-67    DOI: DOI:10.13523/j.cb.20160309
    
In vitro Refolding of an Outer Membrane Protein from a Psychrotrophic Shewanella sp.
XUAN Huan-ling, LI Jing, LUO Feng, DAI Xian-zhu
Research Center of Bioenergy and Bioremediation, College of Resources and Environment, Southwest University, Chongqing 400716, China
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Abstract  

Objective: To investigate cold-adapatation mechanisms of outer membrane protein folding of psychrotrophic Gram-negative bacterium by constructing the in vitro refolding system. Methods: The outer membrane protein Omp74 from a psychrotrophic bacterial Shewanella sp. was overexpressed as inclusion body in E. coli, purified, dissolved in surfactant or high concentrations of urea, and refolded with non-ionic surfactant. OmpA from E. coli was used as a control from mesotrophic bacteria. Results: Being different from OmpA, the refolding of Omp74 was not sensitive to temperature, and low concentration of sarkosyl facilitated the folding of wild type Omp74 but showed no effect on OmpA folding; C-terminal periplasmic domain inhibited the folding; complete refolding this recombinant OMP in vitro in 1%DDM mixed with 0.4% sarkosyl.



Key wordsOuter membrane protein      Psychrotrophic bacteria      Surfactant      In vitro folding     
Received: 06 September 2015      Published: 04 January 2016
ZTFLH:  Q816  
Cite this article:

XUAN Huan-ling, LI Jing, LUO Feng, DAI Xian-zhu. In vitro Refolding of an Outer Membrane Protein from a Psychrotrophic Shewanella sp.. China Biotechnology, 2016, 36(3): 61-67.

URL:

https://manu60.magtech.com.cn/biotech/DOI:10.13523/j.cb.20160309     OR     https://manu60.magtech.com.cn/biotech/Y2016/V36/I3/61

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