Abstract

Article info Vol. 7  No. 2   pp.  47 ~ 51
Title Crystallization and preliminary X-ray diffraction analysis of Thioredoxin from the feather-degrading thermophile Fervidobacterium islandicum AW-1
Authors Immanuel Dhanasingh and Sung Haeng Lee*
Institutions Department of Cellular and Molecular Medicine, Chosun University School of Medicine, Gwangju 61452, Korea *Correspondence: sunglee@chosun.ac.kr
Abstract Comparative genomics of the order Thermotogales revealed that the feather-degrading thermophilic eubacterium Fervidobacterium islandicum AW-1 has more extensive redox system comprising of enzymes thioredoxin (Trx) and thioredoxin reductase (TrxR) than non-feather degrading Fervidobacterium spp. Therefore, thioredoxin from F.islandicum AW-1 (FiTrx) likely plays a key role in the reduction of disulfide bridges present in feather keratin. In order to reveal the insights of the electron transfer between feather keratin and thioredoxin systems in F.islandium AW-1, the preliminary structural study of FiTrx was implemented. The FiTrx gene was cloned, expressed, purified, and crystallized. FiTrx crystallized in 0.005M cobalt(II) chloride hexahydrate, 0.005 M nickel(II) chloride hexahydrate, 0.005 M cadmium chloride hydrate, 0.005 M magnesium chloride hexahydrate, 0.1 M HEPES pH 7.5 and 12% w/v PEG3350, diffracted X-ray to a 1.25 Å resolution and belonged to the space group P212121, with the unit cell parameters a = 35.81, b = 60.26, and c = 87.64 Å. The asymmetric unit of the crystal had two molecules with a calculated Mathew’s coefficient of 2.15 Å3 Da–1 and a solvent content of 42.79%.