Vapor batch crystallization and preliminary X-ray crystallographic analysis of a cold-active endo-β-1,4-glucanase that was produced through the cold temperature protein expression OTHER

Title
Vapor batch crystallization and preliminary X-ray crystallographic analysis of a cold-active endo-β-1,4-glucanase that was produced through the cold temperature protein expression
Author(s)
안영준; 김민규; 송정민; 강미혜; 이윤호; 차선신
KIOST Author(s)
Young, Jun An(안영준)Lee, Youn Ho(이윤호)
Publication Year
2015-09
Abstract
The CaCel gene product from Antarctic springtail Cryptopygus antarcticus (CaCel) belongs to the glycoside hydrolasefamily 45 (GH45) type endo-β-1,4-glucanase. Since the production of soluble recombinant CaCel was not successful at thetemperature range of 15-37oC, we further lowered the expression temperature. The Escherichia coli Rosetta-gami2 (DE3)strain harbouring an expression vector including the CaCel gene was cultured at 10oC. Due to the extremely low growthrate, the induction time was expanded to 9 days and the 18-liter culture volume was necessary to get enough solubleprotein for crystallization. Crystals of CaCel were grown in droplets under Al’s Oil that allows vapor diffusion. In spite ofsmall size, the crystal of CaCel, which belonged to the space group P3121, with unit-cell parameters a = 73.57, b = 83.93,c = 163.77 Å, diffracted to 2.6 Å resolution.
ISSN
2288-6982
URI
https://sciwatch.kiost.ac.kr/handle/2020.kiost/2413
Bibliographic Citation
Biodesign, v.3, no.3, pp.138 - 142, 2015
Publisher
한국구조생물학회
Keywords
.
Type
Article
Language
English
Publisher
한국구조생물학회
Related Researcher
Research Interests

Marine Molecular Genetics,Marine Ecology and Evolution,해양분자유전,해양생태및진화

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