Abstract
A genomic DNA fragment, encoding a thermotolerant β-glucosidase, of the obligate anaerobe Thermotoga petrophila RKU-1 was cloned after PCR amplification into Escherichia coli strain BL21 CodonPlus. The purified cloned enzyme was a monomeric, 51.5 kDa protein (by SDS-PAGE) encoded by 1.341 kb gene. The estimated K m and V max values against p-nitrophenyl-β-D-glucopyranoside were 2.8 mM and 42.7 mmol min−1 mg−1, respectively. The enzyme was also active against other p-nitrophenyl substrates. Possible catalytic sites involved in hydrolyzing different p-nitrophenyl substrates are proposed based on docking studies of enzyme with its substrates. Because of its unique characters, this enzyme is a potential candidate for industrial applications.
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This work was supported by a Grant No. 27(54)/2007-DSA (P&D) from the Ministry of Science and Technology, Pakistan. We are very grateful to Prof. Dr. David B. Wilson for critically reading the manuscript.
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Ikram Ul Haq and Mahmood Ali khan contributed equally to this study.
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Haq, I.U., Khan, M.A., Muneer, B. et al. Cloning, characterization and molecular docking of a highly thermostable β-1,4-glucosidase from Thermotoga petrophila . Biotechnol Lett 34, 1703–1709 (2012). https://doi.org/10.1007/s10529-012-0953-0
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DOI: https://doi.org/10.1007/s10529-012-0953-0