Preliminary X-ray crystallographic analysis of Cryptopygus antarticus mannanase and cellulase that were produced at cold temperature in Escherichia coli

Title
Preliminary X-ray crystallographic analysis of Cryptopygus antarticus mannanase and cellulase that were produced at cold temperature in Escherichia coli
Author(s)
나정현; 차선신
Alternative Author(s)
나정현; 차선신
Publication Year
2015-11-20
Abstract
Biochemical, biophysical, and structural information on proteins provide opportunity for understand their biological functions. One of the limitations to the protein study is the difficulty in obtaining highly purified soluble protein. Protein expression in Escherichia coli (E.coli) at 15-25 ℃ is used to overcome this solubility limitation. However, many recombinant proteins are insolubly expressed even at those low temperatures. In this study, we show that recombinant proteins can be expressed as soluble forms at 6-10 ℃ without cold adapted chaperon systems. By using E. coli Rosetta-gami2(DE3), we obtained 1.8 and 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 L culture grown at 6 and 10 ℃, respectively. These proteins are also suitable for X-ray crystallography. Crystals of CaMan and CaCel diffracted to 2.4 Å and 2.6 Å resolution, respectively. Consequently, E. coli cultivation at 6– 10 ℃ is an effective method for overcoming a major hurdle of the inclusion body formation.ein expression in Escherichia coli (E.coli) at 15-25 ℃ is used to overcome this solubility limitation. However, many recombinant proteins are insolubly expressed even at those low temperatures. In this study, we show that recombinant proteins can be expressed as soluble forms at 6-10 ℃ without cold adapted chaperon systems. By using E. coli Rosetta-gami2(DE3), we obtained 1.8 and 0.9 mg of Cryptopygus antarticus mannanase (CaMan) and cellulase (CaCel) from 1 L culture grown at 6 and 10 ℃, respectively. These proteins are also suitable for X-ray crystallography. Crystals of CaMan and CaCel diffracted to 2.4 Å and 2.6 Å resolution, respectively. Consequently, E. coli cultivation at 6– 10 ℃ is an effective method for overcoming a major hurdle of the inclusion body formation.
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/25037
Bibliographic Citation
제 27차 방사광이용자연구발표회, pp.173, 2015
Publisher
포항가속기연구소
Type
Conference
Language
English
Publisher
포항가속기연구소
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