Molecular and functional characterization of unique thermo-halophilic thioredoxin from the metagenome of an exotic environment
AbdelWahed, Mohamed G; Abdel-Badiea, Elham; Ouf, Amged; Sayed, Ahmed A;
Abstract
The lower convective layer (LCL) at Atlantis II brine pool of the Red sea represents one of the exceptional, unique ecosystems. Thioredoxin is a multi-functional antioxidant redox protein that has a crucial role in various vital cellular processes. In the current study, a functional metagenomics approach was used to isolate and characterize thioredoxin from the LCL of Atlantis II Deep brine pool (Trx-ATII). From the metagenomic DNA of the LCL, the thioredoxin gene was directly retrieved and sequenced. Sequence analysis showed that the gene belonged to thioredoxin-like superfamily with classical Trx motif (-CXXC-). Phylogenetic analysis revealed that Trx-ATII was closely related to Trx of Prochlorococcus marinus with a maximum identity of 86%. Successfully, Trx-ATII was cloned and expressed in E. coli, where the purified protein had M.wt of 16 kDa. Characterization studies revealed that Trx-ATII protein is halophilic; can tolerate up to 2.5 M NaCl and thermostable, where 90% of its activity was retained at 60 °C. Trx-ATII can reduce both DTNB and insulin disulfide- containing substrates. In conclusion, a unique thioredoxin protein was isolated from a harsh environment that can maintain its activity under extreme conditions of salinity and temperature as a promising redox protein for biotechnological applications.
Other data
Title | Molecular and functional characterization of unique thermo-halophilic thioredoxin from the metagenome of an exotic environment | Authors | AbdelWahed, Mohamed G; Abdel-Badiea, Elham ; Ouf, Amged; Sayed, Ahmed A | Keywords | Brine pools; Halotolerance; Metagenomics; Thermal analysis; Thioredoxin | Issue Date | 15-Jun-2020 | Publisher | ELSEVIER | Journal | International journal of biological macromolecules | ISSN | 0141-8130 | DOI | 10.1016/j.ijbiomac.2020.03.011 | PubMed ID | 32142845 | Web of science ID | WOS:000531080800077 |
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