Preliminary X-ray crystallographic analysis of Cryptopygus antarticus mannanase and cellulase that were produced through the cold temperature protein expression

Title
Preliminary X-ray crystallographic analysis of Cryptopygus antarticus mannanase and cellulase that were produced through the cold temperature protein expression
Author(s)
나정현; 차선신
Alternative Author(s)
나정현; 차선신
Publication Year
2015-11-06
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/25114
Bibliographic Citation
2015년 한국결정학회 학술대회, pp.41, 2015
Publisher
한국결정학회
Type
Conference
Language
English
Publisher
한국결정학회
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