Open AccessExpression, purification, crystallization and preliminary crystallographic study of a potential metal‐dependent hydrolase with cyclase activity from Thermoanaerobacter tengcongensis
Author(s) -
Liu Sen,
Wu Guangteng,
Huang Qichen,
Lai Luhua,
Tang Youqi,
Unno Hideaki,
Kusunoki Masami
Publication year - 2005
Publication title -
acta crystallographica section f
Language(s) - English
Resource type - Journals
ISSN - 1744-3091
DOI - 10.1107/s1744309104029392
Subject(s) - escherichia coli , hydrolase , thermophile , crystallization , molecular replacement , crystallography , chemistry , accession number (library science) , strain (injury) , resolution (logic) , genbank , recombinant dna , crystal structure , biochemistry , gene , enzyme , biology , organic chemistry , anatomy , artificial intelligence , computer science
The putative metal‐dependent hydrolase gene TTE1006 from Thermoanaerobacter tengcongensis strain MB4 T (T = type strain; Genbank accession No. AE008691) was heterologously expressed in Escherichia coli . The 205‐amino‐acid gene product was purified and crystallized. The crystal used for data collection belongs to space group P 2 1 , with unit‐cell parameters a = 85.2, b = 62.1, c = 172.4 Å, β = 104.2°. Using a synchrotron‐radiation source, the resolution limit of the data reached 1.87 Å. Eight molecules were estimated to be present in the asymmetric unit, with a solvent content of 48%. Structure determination is ongoing using the multiple‐wavelength anomalous diffraction (MAD) method and also the molecular‐replacement (MR) method.